Samples separated on polyacrylamide or agarose gels were incubated in transfer buffer (48 mM Tris-HCl, 39 mM glycine, 0.0375% SDS, 20% methanol) and then transferred to a nitrocellulose membrane (Protran; Schleicher and Schuell) in a semidry electroblotter (SD cell, BioRad) (1 h, 200 mA). Blots were blocked with phosphate-buffered saline (PBS) containing 5% nonfat dry milk and incubated with a primary rabbit anti-VP2 antibody diluted in blocking solution (2 h). Membranes were washed and incubated with a horseradish peroxidase-conjugated goat anti-rabbit antibody (GE Healthcare) (1 h). Membranes were developed with ECL chemiluminescence reagent (GE Healthcare).
Horseradish peroxidase conjugated goat anti rabbit antibody
Manufactured by GE Healthcare
Sourced in United Kingdom
Horseradish peroxidase-conjugated goat anti-rabbit antibody is a secondary antibody used in various immunoassay techniques. It contains a horseradish peroxidase enzyme that is conjugated to a goat-derived antibody specific to rabbit immunoglobulins. This product can be used to detect and quantify the presence of rabbit primary antibodies in samples.
Automatically generated - may contain errors
Lab products found in correlation
Ecl chemiluminescence reagent, by GE Healthcare (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
A9044, by Merck Group (1 mentions)
Ecl plus detection system, by GE Healthcare (1 mentions)
Horseradish peroxidase conjugated goat anti mouse antibody, by Merck Group (1 mentions)
Na934, by GE Healthcare (1 mentions)
Rabbit anti creb, by Cell Signaling Technology (1 mentions)
Polyvinylidene fluoride (pvdf), by GE Healthcare (1 mentions)
Enhanced chemiluminescence ecl substrate, by GE Healthcare (1 mentions)
Rabbit anti pcreb, by Cell Signaling Technology (1 mentions)
4 protocols using horseradish peroxidase conjugated goat anti rabbit antibody
1
Western Blot Analysis of VP2 Protein
2
Acetate-Induced ERK1/2 Activation in M2 Macrophages
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M2 polarized bone marrow derived macrophages were cultured in serum-free RPMI 1640 (Gibco) for 3 h, and then stimulated with acetate (10mM;Wako). After 10 min, cells were lysed in TNE buffer and centrifuged at 14,000 g for 20 min at 4°C. The supernatants were resolved by SDS gel electrophoresis and blotted onto a nitrocellulose membrane. Primary antibodies used were follows: ERK1/2 (Cell signaling; rabbit, 1:1,000) and phosphorylated ERK1/2 (Cell signaling; rabbit, 1:1,000). The secondary antibody used was a horseradish peroxidase-conjugated goat anti-rabbit antibody (GE Healthcare; 1:2000). Immunoreactive bands were visualized using an enhanced chemiluminescence detection system. Image J (National Institutes of Health) was used to quantify the integrated density of each band.
3
Western Blot Analysis of Galectin-7 Protein
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Protein extraction and Western blot analysis were done as previously described [14 (link)]. Galectin-7 protein was detected using the rabbit polyclonal anti-galectin-7 antibody (1:3000; [7 (link)]) and horseradish peroxidase-conjugated goat anti-rabbit antibody (1:10000, GE Healthcare NA934, Little Chalfont, Buckinghamshire, United Kingdom). ß-tubulin protein was detected with a mouse monoclonal anti-ß-tubulin antibody (1:8000, GE Healthcare N357) and horseradish peroxidase-conjugated goat anti-mouse antibody (1:15000, A9044, Sigma-Aldrich, St. Louis, MO). Signals were revealed using the ECL Plus detection system (GE Healthcare RPN2132).
4
Western Blot Analysis of pCREB in Rat Cortical Neurons
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Rat cortical neurons were lysed in Laemmli sample buffer (2% SDS, 10% glycerol, 60 mM Tris-Cl, 0.01% bromophenol blue) and proteins were resolved via sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), followed by transfer onto polyvinylidene fluoride (PVDF, GE Healthcare, United Kingdom) membranes. Blots were blocked in 5% fat-free milk in 0.05% Tris Buffered Saline-Tween 20 (TBS-T) and incubated in primary antibodies in 0.05% TBS-T, overnight at 4°C. Primary antibodies included rabbit anti-pCREB (1:500; Cell Signaling Technology, MA, United States) and rabbit anti-CREB (1:1000, Cell Signaling Technology). Blots were washed 3–5 times and incubated with a 1:5000 dilution of horseradish peroxidase-conjugated goat anti-rabbit antibody (GE Healthcare, United Kingdom) for 1 h. Protein-antibody complexes were detected on X-ray films following addition of Enhanced Chemiluminescence (ECL) substrate (GE Healthcare, United Kingdom). The relative density of the pCREB and CREB bands was quantitated using ImageJ software (NIH, United States), and was represented as a pCREB/CREB ratio.
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