Original iBlot® Gel Transfer Device

Find resources for the original iBlot® Gel Transfer Device, including details and performance data for the iBlot® Western Detection Kits as well as application notes, FAQs, literature citations, ordering information, and more.

 

Q: I saw that the iBlot® 2 Dry Blotting System is now available. Can I still buy consumables for the original iBlot® Gel Transfer Device?

A: Yes, they are still available. You can find them in the product list on this page.

Q: Is the iBlot® 2 Gel Transfer Device compatible with original iBlot® Transfer Stacks?

A: No. Do not use iBlot® Transfer Stacks in the iBlot® 2 Gel Transfer Device, and do not mix components between iBlot® Transfer Stacks and iBlot® 2 Transfer Stacks.

Q: Is there a limitation on the thickness of gels that can be used with the iBlot® Dry Blotting System?

A: The iBlot® Dry Blotting System has been tested to efficiently transfer protein from gels ranging in thickness from 1 mm to 3 mm. We have not tested gels thicker than 3 mm because they are rarely used for SDS-PAGE.

Q: I want to conduct western transfer with mini gels (8 x 8 cm), but I don’t have iBlot® Transfer Stacks (Mini). Can I use iBlot® Transfer Stacks (Regular) to transfer my mini gels?

A: It is best not to transfer a single mini gel on a regular-sized transfer stack. Although in most cases the transfer will be fine, empty spaces on the transfer stack that are not in direct contact with a gel could potentially cause distortions across the whole surface of the membrane, including the portion in contact with the gel. It is best to have more than 50% of a membrane in contact with the gel, if possible.

Q: How can we be more environmentally responsible?

A: The plastic in the iBlot® Dry Blotting System stack packaging is polyethylene terephthalate (PET) and can be recycled.

Q: Are the copper electrodes in the transfer stacks recyclable?

A: Yes, and we have established a recycling program available for customers in North America only.  The electrodes are copper-coated nylon, and the amount of copper left in the electrode after a transfer is rather small. There is 0.6 grams of copper per sheet before transfer, and even less after transfer.

Q: Sometimes there is green discoloration on the blot around the gel. What is this, and does it affect the results?

A: The green discoloration is copper deposits from the iBlot® Transfer Stacks, and it does not affect the results. To minimize this effect, shake excess water off the filter paper and buffer from the gel before placing each on the stack. The current formulation of stacks minimizes the green discoloration.

Q: Do the PVDF iBlot® Transfer Stacks (0.2 µm, non-autofluorescing) require activation prior to use?

A: No. The PVDF membrane comes preactivated. You just need to open the transfer stack with membrane, place the separation gel on top of the membrane, and apply one layer of moistened filter paper to run (the same as with the nitrocellulose stacks).

Q: Can iBlot® Transfer Stacks be used more than once?

A: No. The transfer stacks have a finite amount of ions to drive the transfer and are depleted after a single use.

Q: What is the shelf life of both the nitrocellulose and PVDF iBlot® Transfer Stacks?

A: The minimum guaranteed shelf life for iBlot® Transfer Stacks is 2 months. Depending on when you purchase the transfer stack, shelf life will be 2–8 months.

Q: Is it possible to substitute the membrane from the iBlot® Transfer Stack with my specialized membrane?

A: In theory, you can replace the membrane provided in your iBlot® Transfer Stack with any membrane that is compatible with western blotting. To do this, cut the alternative membrane to match the size of your gel, and wet the membrane. Then either place the alternative membrane on top of the integrated membrane, or carefully remove the integrated membrane from the gel matrix with forceps and replace it with the new membrane. Note that Life Technologies only supports the use of iBlot® Transfer Stacks when they are used with the provided instructions.

Q: Occasionally my western blots have high background. What do you recommend?

A: This may be a result of insufficient blocking or nonspecific binding. We suggest trying our WesternBreeze® Blocker/Diluent. We have been using it with good success. Additionally, you should optimize primary and secondary antibody concentrations as generally recommended for any new blotting technique. Many cases of high background can be resolved by further diluting one or both antibody preparations.

Q: How can I get better transfer of high molecular weight proteins?

A: Proteins larger than ~150 kDa migrate more slowly than smaller proteins. Therefore, more time is required to transfer them from gel to membrane. We recommend extending the transfer time by 8–10 minutes for optimal transfer of proteins >150 kDa using the iBlot® Dry Blotting System.

To enhance transfer efficiency, we also recommend adding an equilibration (gel-soaking) step between electrophoresis and western transfer and using NuPAGE® Novex 3-8% Tris-acetate gels for electrophoresis. We have an application note available, titled “Transferring Large and Small Proteins Using the iBlot® 7-Minute Blotting System”. To download the pdf, see Application notes tab on this page.

Q: What causes empty spots on my membrane after transfer?

A: The iBlot® Dry Blotting System is similar to conventional transfer methods in that air bubbles between the gel and the membrane will prevent protein transfer. Be sure to remove all air bubbles between the gel and membrane before starting the transfer, using the blotting roller supplied with the iBlot® Dry Blotting System.

Q: Is there a stripping protocol for the iBlot® Dry Blotting System?

A: A conventional stripping protocol using 0.1 M glycine, pH 2, works with polyclonal antibodies.

Q: Does the iBlot® Dry Blotting System work with native or native-blue gels?

A: Yes, we have an application note available, titled, “Western Blotting NativePAGE™ Novex® Bis-Tris gels Using the iBlot® 7-Minute Blotting System’. To download the pdf, see Application notes tab on this page.

Q: Do I need to have an iBlot® Dry Blotting System to use an iBlot® Western Detection Kit?

A: Yes. The iBlot® Western Detection Kits use the iBlot® system to produce an electrical field that accelerates interactions between antibodies and blocking reagents with membrane-bound antigens. iBlot® Western Detection Kits require an iBlot® Dry Blotting System with the P9 program (available in software versions 2.9.5 and higher). If you have an older iBlot® Dry Blotting System, the firmware is freely available for download. See the Instrument Registration page to find out how.

Q: Can iBlot® Western Detection Stacks be used more than once?

A: No. The detection stacks have a finite amount of ions and are depleted after a single use, which includes the three detection steps: blocking, primary antibody, and secondary antibody.

Q: Should I change the detection stacks for each step?

A: No. A single set of stacks is used for all three steps of the western detection.

Q: Does it matter what method I use to transfer protein to my western blot membrane?

A: No, it does not matter. iBlot® Western Detection Kits are compatible with western blots created using wet or semi-dry methods. You can use one regular-sized membrane (13.5 cm x 8 cm) or 1–2 mini-sized membranes (8 x 8 cm).

Q: How many reactions can I perform at one time?

A: iBlot® Western Detection Stacks come in two sizes: mini and regular; they are supplied with assay spacers to create partitions for analysis using more than one set of antibodies. With the mini size you can perform detection on one mini-sized membrane or two halves of a mini-sized membrane. With the regular size you can perform detection on two mini-sized membranes or 4 quarters of a regular-sized membrane (4 x 8 cm).

Q: Sometimes I see high background on the membrane after detection. How can I eliminate it?

A: High background can be caused by nonspecific antibody binding. To eliminate it, use a lower concentration of the secondary antibody (as specified in the manual). In addition, using nitrocellulose rather than PVDF membranes will help to minimize background.

Q: Sometimes my molecular weight (MW) markers are no longer visible after my detection is completed. Does this mean my proteins are also gone?

A: Definitely not. MW markers are usually prestained, and the stains are more highly charged than typical cellular proteins. Applying an electrical field in the western detection procedure can drive the MW markers right through the membrane onto the bottom stack.

Cellular proteins, however, are less charged than the MW markers, and at the detection stage they no longer contain any SDS. The result is that the cellular proteins stay immobilized on the membrane during western detection using the iBlot® system.

Q: Can I use different antibodies or different dilutions of the same antibody in the procedure?

A: Yes. iBlot® Western Detection Kits are supplied with reusable spacers, expressly for this purpose. Use spacers to separate membrane sections for detection with different antibody solutions at the same time. A detailed protocol is provided in the user manual.

Q: What membrane type should I use with the iBlot® Western Detection Kits? Is one type preferred over another?

A: You can use nitrocellulose or PVDF membranes with the iBlot® Western Detection Kits. However, we highly recommend using nitrocellulose rather than PVDF because less background is seen with nitrocellulose membranes. This results in more sensitive detection.

Q: Can I use the iBlot® Western Detection Kits on membranes with previously transferred proteins?

A: Yes, as long as you activate the membrane before you start the detection procedure. Completely wet nitrocellulose membranes using water. Activate PVDF membranes by immersing in 100% methanol for several seconds until the membrane is completely wet, then rinse with water. Then proceed with the iBlot® Western Detection Kit.

Q: What substrate is used with the kits?

A: The substrate used for the iBlot® Western Detection Kits reacts with alkaline phosphatase (AP). Therefore, horseradish peroxidase (HRP) substrates will not work for this kit. Other conjugates and substrates of AP cannot be used with iBlot® Western Detection Kits. The reagents supplied with the kits have been optimized to obtain the best results.

Q: How is it possible to finish the entire immunodetection in just 30 minutes?

A: Native antibodies typically have some negative charge at neutral pH, which allows their electrophoretic migration in low voltage from the carrier matrix onto the membrane. The force of the electric field is counterbalanced by their affinity for antigen, so that binding antibodies are “captured” while unbound antibodies flow through the blocked membrane and onto the bottom stack. Basically, the iBlot® Dry Blotting System electrophoretically focuses antibodies to greatly accelerate their interaction rate with immobilized antigens.

Q: Are the kits compatible with the Li-Cor® system?

A: Currently, the iBlot® Western Detection Kits use a green matrix, and unfortunately this can create background fluorescence that isn’t compatible with the Li-Cor® Odyssey scanner.

Q: How can I improve the signal intensity?

A: The signal intensity is primarily controlled by the concentration of the secondary antibody; nonspecific binding can result when the concentration is too high, and poor binding to the primary antibody can result when the solution is too dilute. The next option is to modify the primary antibody concentration, which controls the number of antibody molecules that bind to the antigen, and thus the signal and background levels. (See the manual for further details.)

Q: Can the primary antibody solution be reused?

A: No. The primary antibody solution cannot be reused after it has been applied to the matrix.

Q: What concentration of primary antibody should I use with the iBlot® Western Detection Kits?

A: Use twice the primary antibody concentration used in typical western blot detection procedures. Note that the overall amount of the primary antibody used is the same as in typical traditional methods, because only half the volume of primary antibody solution is needed with the iBlot® Western Detection System.

Q: What is the shelf life of the iBlot® Western Detection Stacks?

A: The shelf life is 6 months, at room temperature.

Q: What is the shelf life of the iBlot® Western Detection Reagents?

A: The shelf life is 12 months, at 4°C.

Q: I have reagents left but the detection stacks have expired. Can I purchase only the iBlot® Western Detection Stacks?

A: Yes. To use your reagents with new stacks, you can purchase the stacks only: iBlot® Western Detection Stacks, regular size and iBlot® Western Detection Stacks, mini size.

2011

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Huang-Doran I et al. 2011. Genetic defects in human pericentrin are associated with severe insulin resistance and diabetes. Diabetes 60:925–935.

Ibebunjo C et al. 2011. Voluntary running, skeletal muscle gene expression, and signaling inversely regulated by orchidectomy and testosterone replacement. Am J Physiol Endocrinol Metab 300:E327–E340.

Iben JR et al. 2011. Comparative whole genome sequencing reveals phenotypic tRNA gene duplication in spontaneous Schizosaccharomyces pombe La mutants. Nucleic Acids Res 2011 Feb. 11. [Epub ahead of print]

Iqbal M et al. 2011. Corticosteroid regulation of p-glycoprotein in the developing blood-brain barrier. Endocrinology 152:1067–1079.

Liew ATF et al. 2011. A simple plasmid-based system that allows rapid generation of tightly controlled gene expression in Staphylococcus aureus. Microbiology 157:666–676.

Maruri-Avidal L et al. 2011. Participation of vaccinia virus L2 protein in the formation of crescent membranes and immature virions. J Virol 85:2504–2511.

McGinnes LW et al. 2011. Assembly and immunological properties of newcastle disease virus-like particles containing the respiratory syncytial virus F and G proteins. J Virol 85:366–377.

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Shtanko O et al. 2011. ALIX/AIP1 is required for NP incorporation into mopeia virus Z-induced virus-like particles. J Virol 85:3631–3641.

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Xu Q et al. 2011. Spinal phosphinositide 3-kinase–Akt–mammalian target of rapamycin signaling cascades in inflammation-induced hyperalgesia. J Neurosci 31:2113–2124.

2010

Adlard PA et al. 2010. Cognitive loss in zinc transporter-3 knock-out mice: a phenocopy for the synaptic and memory deficits of Alzheimer’s disease? J Neurosci 30:1631–1636.

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Bach-Ngohou K et al. 2010. Enteric glia modulate epithelial cell proliferation and differentiation through 15-deoxy-Δ12,14-prostaglandin J2. J Physiol 588:2533–2544.

Baeyens N et al. 2010. Identification and functional implication of a Rho kinase-dependent moesin-EBP50 interaction in noradrenaline-stimulated artery. Am J Physiol Cell Physiol 299:C1530–C1540.

Barry C et al. 2010. Features of a spatially constrained cystine loop in the p10 FAST protein ectodomain define a new class of viral fusion peptides. J Biol Chem 285:16424–16433.

Bergeron E et al. 2010. Crimean-Congo hemorrhagic fever virus-encoded ovarian tumor protease activity is dispensable for virus RNA polymerase function. J Virol 84:216–226.

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Bryan MA et al. 2010. Genetic immunization converts the Trypanosoma cruzi B-cell mitogen proline racemase to an effective immunogen. Infect Immunol 78:810–822.

Checkley MA et al. 2010. P-Body components are required for Ty1 retrotransposition during assembly of retrotransposition-competent virus-like particles. Mol Cell Biol 30:382–398.

Chen J et al. 2010. Protein-disulfide isomerase-associated 3 (Pdia3) mediates the membrane response to 1,25-dihydroxyvitamin D3 in osteoblasts. J Biol Chem 285:37041–37050.

Cortes R et al. 2010. Influence of heart failure on nucleocytoplasmic transport in human cardiomyocytes. Cardiovasc Res 85:464–472.

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Fredriksen L et al. 2010. Cell wall anchoring of the 37-kilodalton oncofetal antigen by Lactobacillus plantarum for mucosal cancer vaccine delivery. Appl Envir Microbiol 76:7359–7362.

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Gerus M et al. 2010. Evolutionarily conserved function of RRP36 in early cleavages of the pre-rRNA and production of the 40S ribosomal subunit. Mol Cell Biol 30:1130–1144.

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Graziani A et al. 2010. Cell-cell contact formation governs Ca2+ signaling by TRPC4 in the vascular endothelium: evidence for a regulatory TRPC4-β-catenin interaction. J Biol Chem 285:4213–4223.

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Jian MJ et al. 2010. Ins(1,4,5)P3 interacts with PIP2 to regulate activation of TRPC6/C7 channels by diacylglycerol in native vascular myocytes. J Physiol 588:1419–1433.

Jiang XS et al. 2010. Activation of Rho GTPases in Smith-Lemli-Opitz syndrome: pathophysiological and clinical implications. Hum Mol Genet 19:1347–1357.

Jiang XS et al. 2010. quantitative proteomics analysis of inborn errors of cholesterol synthesis: identification of altered metabolic pathways in DHCR7 and SC5D deficiency. Mol Cell Proteomics 9:1461–1475.

Kahler D et al. 2010. Proteomics out of the archive: two-dimensional electrophoresis and mass spectrometry using HOPE-fixed, paraffin-embedded tissues. J Histochem Cytochem 58:221–228.

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Karlsson JE et al. 2010. High-activity P-glycoprotein, multidrug resistance protein 2, and breast cancer resistance protein membrane vesicles prepared from transiently transfected human embryonic kidney 293-Epstein-Barr virus nuclear antigen cells. Drug Metab Dispos 38:705–714.

Kelley R et al. 2010. Tubular cell-enriched subpopulation of primary renal cells improves survival and augments kidney function in rodent model of chronic kidney disease. Am J Physiol Renal Physiol 299:F1026–F1039.

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Kosaka K et al. 2010. Role of Nrf2 and p62/ZIP in the neurite outgrowth by carnosic acid in PC12h cells. J Biochem 147:73–81.

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Lazo JS et al. 2010. Identifying a resistance determinant for the antimitotic natural products disorazole C1 and A1. J Pharmacol Exp Ther 332:906–911.

Lee JH et al. 2010. Histone deacetylase inhibitor induces DNA damage, which normal but not transformed cells can repair. Proc Natl Acad Sci USA 107:14639–14644.

Lin JJ et al. 2010. Hsp90 directly modulates the spatial distribution of AF9/MLLT3 and affects target gene expression. J Biol Chem 285:11966–11973.

Lopez JE et al. 2010. A novel surface antigen of relapsing fever spirochetes can discriminate between relapsing fever and lyme borreliosis. Clin Vaccine Immunol 17:564–571.

Lucas TM et al. 2010. Pseudotyping incompatibility between HIV-1 and gibbon ape leukemia virus Env is modulated by Vpu. J Virol 84:2666–2674.

Maldonado EN et al. 2010. Free tubulin modulates mitochondrial membrane potential in cancer cells. Cancer Res 70:10192–10201.

Mamedova LK et al. 2010. Tissue expression of angiopoietin-like protein 4 in cattle. J Anim Sci 88:124–130.
Mao Y et al. 2010. Golgi protein 73 (GOLPH2) is a valuable serum marker for hepatocellular carcinoma. Gut 59:1687–1693.

Mbefo MK et al. 2010. Phosphorylation of synucleins by members of the Polo-like kinase family. J Biol Chem 285:2807–2822.

Mendonca MC et al. 2010. Transforming growth factor-β1 regulation of C-type natriuretic peptide expression in human vascular smooth muscle cells: dependence on TSC22D1. Am J Physiol Heart Circ Physiol 299:H2018–H2027.

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Miehe S et al. 2010. The phospholipid-binding protein SESTD1 Is a novel regulator of the transient receptor potential channels TRPC4 and TRPC5. J Biol Chem 285:12426–12434.

Mizutani M et al. 2010. Connective tissue growth factor (CTGF/CCN2) is increased in peritoneal dialysis patients with high peritoneal solute transport rate. Am J Physiol Renal Physiol 298:F721–F733.

Murakami E et al. 2010. Mechanism of activation of PSI-7851 and its diastereoisomer PSI-7977. J Biol Chem 285:34337–34347.

Murawski MR et al. 2010. Newcastle disease virus-like particles containing respiratory syncytial virus G protein induced protection in BALB/c mice, with no evidence of immunopathology. J Virol 84:1110–1123.

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Nishimura H et al. 2010. Cellular localization of sphingosine-1-phosphate receptor 1 expression in the human central nervous system. J Histochem Cytochem 58:847–856.

O’Brien C et al. 2010. Predictive biomarkers of sensitivity to the phosphatidylinositol 3’ kinase inhibitor GDC-0941 in breast cancer preclinical models. Clin Cancer Res 16:3670–3683.

O’Brien NA et al. 2010. Activated phosphoinositide 3-kinase/AKT signaling confers resistance to trastuzumab but not lapatinib. Mol Cancer Ther 9:1489–1502.

Ohishi M et al. 2010. The relationship between HIV-1 genome RNA dimerization, virion maturation and infectivity. Nucleic Acids Res 10:1093.

Parelkar NK et al. 2010. 2,2,2-Trichloroethanol activates a nonclassical potassium channel in cerebrovascular smooth muscle and dilates the middle cerebral artery. J Pharmacol Exp Ther 332:803–810.

Potrykus K et al. 2010. Imprecise transcription termination within Escherichia coli greA leader gives rise to an array of short transcripts, GraL. Nucleic Acids Res 38:1636–1651.

Puel A et al. 2010. Autoantibodies against IL-17A, IL-17F, and IL-22 in patients with chronic mucocutaneous candidiasis and autoimmune polyendocrine syndrome type I. J Exp Med 207:291–297.

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Ranalletta M et al. 2010. Biochemical characterization of cholesteryl ester transfer protein inhibitors. J Lipid Res 51:2739–2752.

Rodriguez HM et al. 2010. Modulation of lysyl oxidase-like 2 enzymatic activity by an allosteric antibody inhibitor. J Biol Chem 285:20964–20974.

Roux J et al. 2010. Transforming growth factor β1 inhibits cystic fibrosis transmembrane conductance regulator-dependent cAMP-stimulated alveolar epithelial fluid transport via a phosphatidylinositol 3-kinase-dependent mechanism. J Biol Chem 285:4278–4290.

Sakana A et al. 2010. Rab13 regulates neurite outgrowth in PC12 cells through its effector protein, JRAB/MICAL-L2. Mol Cell Biol 30:1077–1087.

Sakr S et al. 2010. Lon protease quality control of presecretory proteins in Escherichia coli and its dependence on the SecB and DnaJ (Hsp40) chaperones. J Biol Chem 285:23506–23514.

Sayeedur M et al. 2010. Functional characterization of a phospholipase A2 homolog from Rickettsia typhi. J Bacteriol 192:3294–3303.

Schnitzler AC et al. 2010. BMP9 (bone morphogenetic protein 9) induces NGF as an autocrine/paracrine cholinergic trophic factor in developing basal forebrain neurons. J Neurosci 30:8221–8228.

Sharbeen G et al. 2010. Incorporation of dUTP does not mediate mutation of A:T base pairs in Ig genes in vivo. Nucleic Acids Res 38:8120–8130.

Spicakova T et al. 2010. Expression and silencing of the microtubule-associated protein Tau in breast cancer cells. Mol Cancer Ther 9:2970–2981.

Srirajaskanthan R et al. 2010. Identification of Mac-2-binding protein as a putative marker of neuroendocrine tumors from the analysis of cell line secretomes. Mol Cell Proteomics 9:656–666.

Tahir SK et al. 2010. Identification of expression signatures predictive of sensitivity to the Bcl-2 family member inhibitor ABT-263 in small cell lung carcinoma and leukemia/lymphoma cell lines. Mol Cancer Ther 9:545–557.

Tsuji H et al. 2010. Xenografted human amniotic membrane–derived mesenchymal stem cells are immunologically tolerated and transdifferentiated into cardiomyocytes. Circ Res 106:1613–1623.

Twu YC et al. 2010. Phosphorylation status of transcription factor C/EBPα determines cell-surface poly-LacNAc branching (I antigen) formation in erythropoiesis and granulopoiesis. Blood 115:2491–2499.

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Ueta M et al. 2010. Expression of the interleukin-4 receptor αin human conjunctival epithelial cells. Br J Ophthalmol 94:1239–1243.

Wallin JJ et al. 2010. Nuclear phospho-Akt increase predicts synergy of PI3K inhibition and doxorubicin in breast and ovarian cancer. Science Translational Medicine 2:48ra66

Wang HX et al. 2010. WNT2 regulates DNA synthesis in mouse granulosa cells through beta-catenin. Biol Reprod 82:865–875.

Wilhelm MT et al. 2010. Isoform-specific p73 knockout mice reveal a novel role for ΔNp73 in the DNA damage response pathway. Genes Dev 24:549–560.

Wilson CG et al. 2010. Liver-specific overexpression of pancreatic-derived factor (PANDER) induces fasting hyperglycemia in mice. Endocrinology 151:5174–5184.

Yagami A et al. 2010. IL-33 mediates inflammatory responses in human lung tissue cells. J Immunol 185:5743–5750.

Yun CS et al. 2010. Pmr, a histone-like protein H1 (H-NS) family protein encoded by the IncP-7 plasmid pCAR1, is a key global regulator that alters host function. J Bacteriol 192:4720–4731.

Zhu JX et al. 2010. SHP-2 phosphatase activity is required for PECAM-1-dependent cell motility. Am J Physiol Cell Physiol 299:C854–C865.

Zhu Y et al. 2010. Phenotypic plasticity of the ovarian surface epithelium: TGF-β1 induction of epithelial to mesenchymal transition (EMT) in vitro. Endocrinology 151:5497–5505.

2009

Albarino CG et al. 2009. Efficient reverse genetics generation of infectious junin viruses differing in glycoprotein processing. J Virol 83:5606–5614.

Besheer J et al. 2009. Interoceptive effects of alcohol require mglu5 receptor activity in the nucleus accumbens. J Neurosci 29:9582–9591.

Blaney EN et al. 2009. Increase in ALK1/ALK5 ratio as a cause for elevated MMP-13 expression in osteoarthritis in humans and mice. J Immunol 182:7937–7945.

Boehme SA et al. 2009. Murine bone marrow-derived mast cells express chemoattractant receptor-homologous molecule expressed on T-helper class 2 cells (CRTh2). Int Immunol 21:621–632.

Bradford BJ et al. 2009. Daily injection of tumor necrosis factor-α increases hepatic triglycerides and alters transcript abundance of metabolic genes in lactating dairy cattle. J Nutr 139:1451–1456.

Brown RA et al. 2009. The R753Q polymorphism abrogates toll-like receptor 2 signaling in response to human cytomegalovirus. Clin Infect Dis 49:e96–e99.

Calandra JM et al. 2009. Selective survival rescue in 15-lipoxygenase-1-deficient retinal pigment epithelial cells by the novel docosahexaenoic acid-derived mediator, neuroprotectin D1. J Biol Chem 284:17877–17882.

Carrozzino F et al. 2009. Inhibition of basal p38 or JNK activity enhances epithelial barrier function through differential modulation of claudin expression. Am J Physiol Cell Physiol 297:C775–C787.

Cecchini S et al. 2009. Evidence of prior exposure to human bocavirus as determined by a retrospective serological study of 404 serum samples from adults in the United States. Clin Vaccine Immunol 16:597–604.

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Chen J et al. 2009. Inhibition of TRPC1/TRPC3 by PKG contributes to NO-mediated vasorelaxation. Am J Physiol Heart Circ Physiol 297:H417–H424.

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Great system for westerns

The iBlot has been very easy to use, fast with very clear results and there are rarely any messes to clean up.

Date: February 5, 2010
Reviewer: Jennifer Walter
Institution: University of Minnesota


 

Fast transfer

It is so nice to have a IBLOT, 7 min, it save my time, also I need not to prepare smelly buffer for transfer

Date: October 30, 2009
Reviewer: Dr. Jane Lu
Institution: University of Kansas, Medical Center


 

Useful and convenient

I remember the days when it took hours to do a western blot. Now with the iBlot, it takes only 7 minutes! You save a ton of time sitting around; instead, you can get on with your experiments/results in less than 10 minutes. It's quick and gives you just as good a blot as any old western blot technique. But I do agree with another reviewer that the device can get a bit hot, therefore "frying" the gel. The gel may get really warm but so far that hasn't affected any of my experiments.

Date: February 5, 2010
Reviewer: Name
Institution: Kalobios


 

Increases blot throughput

The iBlot has been a great addition to the lab. It is very easy to use, transfers very quickly, and is mess-free with its no buffer system. Overall, I have been very happy with it. The only issues I have are when I'm transferring very small proteins (~15 kDa) I have incomplete transfers, for these I still use the Invitrogen blot module. Otherwise, it is a great set up.

Date: February 5, 2010
Reviewer: Carly
Institution: Repligen


 

Fast and convenient

Easy and fast transfers with no need for transfer buffers. Not quite as good with very large molecular weight proteins.

Date: February 5, 2010
Reviewer: Dr. JP Hegarty
Institution: PSU College of Medicine


Western detection kits for use with the iBlot® Gel Transfer Device

  • Fast—complete western detection in less then 25 minutes
  • Flexible—works with mini and midi gels
  • Easy optimization—allows the use of different conditions for different sections of the blot
  • Optimized components—kit contains the reagents and detection stacks needed for western detection; all you supply is the primary antibody

Designed to be compatible with the original iBlot® Gel Transfer Device, iBlot® Western Detection Kits offer complete western detection in less than 25 minutes with sensitivity that is equal to (or better than) conventional protocols for most antibody–antigen pairs. The kits are available with anti-mouse or anti-rabbit secondary antibodies and are compatible with chemiluminescent and chromogenic detection.

 

For Research Use Only. Not for use in diagnostic procedures.