Horseradish peroxidase conjugated secondary antibody against rabbit igg

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Horseradish peroxidase-conjugated secondary antibody against rabbit IgG. This antibody is designed to bind to rabbit immunoglobulin G (IgG) and is conjugated with the enzyme horseradish peroxidase, which can be used for signal detection in various immunoassays.

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Lab products found in correlation

Rabbit polyclonal antibody, by Merck Group (2 mentions) Nitrocellulose membrane, by Merck Group (1 mentions) P erk1 2, by Cell Signaling Technology (1 mentions) Semi dry blot system, by Bio-Rad (1 mentions) Restore western blot stripping buffer, by Thermo Fisher Scientific (1 mentions) Yap taz, by Cell Signaling Technology (1 mentions) Gapdh, by Merck Group (1 mentions) P akt, by Cell Signaling Technology (1 mentions) Erk1 2, by Cell Signaling Technology (1 mentions) Supersignal west femto maximum sensitivity substrate, by Thermo Fisher Scientific (1 mentions) Phosstop phosphatase inhibitor, by Roche (1 mentions) Complete protease inhibitor, by Roche (1 mentions) Horseradish peroxidase conjugated goat anti mouse igg, by Santa Cruz Biotechnology (1 mentions) β actin monoclonal antibody, by Santa Cruz Biotechnology (1 mentions) Ecl chemiluminescence kit, by GE Healthcare (1 mentions) Caspase 8, by Immunoway (1 mentions) Bcl 2, by Immunoway (1 mentions) Bcl2 associated x (bax), by Immunoway (1 mentions) Beclin1, by Immunoway (1 mentions) Caspase3, by Immunoway (1 mentions)

5 protocols using horseradish peroxidase conjugated secondary antibody against rabbit igg

Western Blot Analysis of Apoptosis Signaling

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Total protein was assessed by SDS-polyacrylamide gel electrophoresis under reducing conditions on 12% gels and then was transferred to nitrocellulose membranes using a tank transfer at 80 mV in Tris-glycine buffer containing 20% methanol for 12 h at 4 °C. Nitrocellulose membranes were blocked for 1 h with 5% bovine serum albumin (BSA) at 37 °C and were incubated for 12 h with the following antibodies: RNF11 (1:800), RIPK1 (1:1500), RIPK3 (1:1500), Caspase8 (1:1000), MLKL (1:1000), JNK (1:2000), ERK (1:1000), p38 (1:1000), PhosphoPlus JNK (1:1000), PhosphoPlus ERK (1:2000) and PhosphoPlus p38 (1:1000) at 4 °C. Next, the membranes were incubated with horseradish peroxidase-conjugated secondary antibody against rabbit IgG (1:1500, Santa Cruz, CA, USA) using the ECL kit (Kangweishiji Biotechnology, Beijing, China). The GAPDH content was analyzed as the loading control with rabbit polyclonal antibody (Sigma, USA).

Yang T., Cao C., Yang J., Liu T., Lei X.G., Zhang Z, & Xu S. (2017). miR-200a-5p regulates myocardial necroptosis induced by Se deficiency via targeting RNF11. Redox Biology, 15, 159-169.

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Comprehensive Protein Analysis by Western Blot

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Whole-cell and tissue extracts were prepared using RIPA buffer containing PhosSTOP phosphatase inhibitor (Roche) and cOmplete protease inhibitor (Roche). Proteins were subjected to 10% SDS–PAGE and blotted on a nitrocellulose membrane (Sigma) with a semi-dry blot system (BioRad). Membranes were incubated with the following primary antibodies: p-ERK1/2 (1:2000, Cell Signaling, #4370), ERK1/2 (1:1000, Cell Signaling, #4695), p-AKT (1:2000, Cell Signaling, #4060), AKT (1:1000 Cell Signaling, #9272), YAP/TAZ (1:1000, Cell Signaling, #8418), DDR1 (1:1000, Cell Signaling, #5583) followed by incubation with horseradish peroxidase-conjugated secondary antibody against rabbit IgG (1:5000, Santa Cruz, #sc-2004). GAPDH (1:15000, Sigma, #G9295) was used as loading control. Blots were visualized using SuperSignal™ West Femto Maximum Sensitivity Substrate (ThermoFisher Scientific) and bands were quantified with ImageJ software. If necessary, stripping was performed with Restore Western Blot Stripping Buffer (ThermoFisher Scientific).

Affo S., Nair A., Brundu F., Ravichandra A., Bhattacharjee S., Matsuda M., Chin L., Filliol A., Wen W., Song X., Decker A., Worley J., Caviglia J.M., Yu L., Yin D., Saito Y., Savage T., Wells R.G., Mack M., Zender L., Arpaia N., Remotti H.E., Rabadan R., Sims P., Leblond A.L., Weber A., Riener M.O., Stockwell B.R., Gaublomme J., Llovet J.M., Kalluri R., Michalopoulos G.K., Seki E., Sia D., Chen X., Califano A, & Schwabe R.F. (2021). Promotion of Cholangiocarcinoma Growth by Diverse Cancer-associated Fibroblast Subpopulations. Cancer cell, 39(6), 866-882.e11.

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Western Blot Analysis of HSP-70 in PBMCs

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The expression of HSP-70 in the PBMCs pellets (n = 10; 2 samples per replicate in each group) was analyzed by Western blot technique using the protocol described by Abass, Kamel, Khalifa, Gouda, El-Manylawi, Mehaisen and Mashaly [30 (link)]. Briefly, 40 μg of the total protein was loaded and separated on 12% polyacrylamide gel containing sodium dodecyl sulphate. Separated proteins were then transferred in Tris-glycine buffer containing 20% methanol, to poly-vinylidene difluoride membranes using a tank transfer for 2 h at 300 mA. Skim milk (5%) was used to block membranes for 1 h and incubated overnight at 4 °C with diluted primary anti-rabbit IgG polyclonal antibody against HSP-70 (1:1000; Cell Signaling Technology, Inc., Danvers, MA, USA). After incubation, a horse radish peroxidase conjugated secondary antibody against rabbit IgG (1:1500; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) was added. Monoclonal β-actin antibody (1:1000; Santa Cruz Biotechnology, Inc.) was added and incubated to the membrane to confirm equal loading of samples followed by a horse radish peroxidase conjugated goat anti-mouse IgG (1:1000; Santa Cruz Biotechnology, Inc.). Detection of HSP-70 was then performed using the ECL chemiluminescence kit (GE Healthcare Life Sciences, Amersham Place, Little Chalfont, Buckinghamshire, UK).

Alzarah M.I., Althobiati F., Abbas A.O., Mehaisen G.M, & Kamel N.N. (2021). Citrullus colocynthis Seeds: A Potential Natural Immune Modulator Source for Broiler Reared under Chronic Heat Stress. Animals : an Open Access Journal from MDPI, 11(7), 1951.

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Protein Expression Analysis via Western Blot

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Total protein was assessed with SDS-polyacrylamide gel electrophoresis under reducing conditions on 12% gels and then transferred to nitrocellulose membranes using a tank transfer at 80 mV in Tris-glycine buffer containing 20 % methanol for 2 h. Nitrocellulose membranes were blocked for 2 h with 5 % skim milk at 37°C and incubated for 12 h with diluted primary swine antibody Nrf2 (1:1000), SOD2 (1:1000), HO-1 (1:1000), Gpx1 (1:1000), TrxR1 (1:1000) and Catalase (1:1000) at 4°C. This was then followed incubated with a horseradish peroxidase-conjugated secondary antibody against rabbit IgG (1:1500, Santa Cruz, CA, USA) using the ECL kit (Kangweishiji Biotechnology, Beijing, China). For statistical analysis, a box plot analysis was applied.

Yang T., Zhao Z., Liu T., Zhang Z., Wang P., Xu S., Lei X.G, & Shan A. (2017). Oxidative stress induced by Se-deficient high-energy diet implicates neutrophil dysfunction via Nrf2 pathway suppression in swine. Oncotarget, 8(8), 13428-13439.

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Western Blot Analysis of Protein Signaling

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Total protein was assessed via SDS-polyacrylamide gel electrophoresis under reducing conditions on 12% gels and then was transferred to nitrocellulose membranes via tank transfer at 80 mV in Tris-glycine buffer containing 20% methanol for 12 h at 4°C. Nitrocellulose membranes were blocked for 1 h with 5% bovine serum albumin (BSA) at 37°C and were incubated for 12 h with the following antibodies at 4°C: PI3K (1:500, Immuno Way, China), AKT (1:500, Immuno Way, China), mTOR (1:500, Immuno Way, China), LC3 (1:1000, Immuno Way, China), Beclin1 (1:500, ImmunoWay, China), BCL2 (1:500, ImmunoWay, China), BAX (1:500, ImmunoWay, China), Caspase3 (1:500, ImmunoWay, China), Caspase8 (1:500, ImmunoWay, China). PBST was used to wash the nitrocellulose membranes 4 times, for 15 minutes each time. Next, the membranes were incubated with horseradish peroxidase-conjugated secondary antibody against rabbit IgG (1:5000, Santa Cruz, CA, USA) using an ECL kit (Kangweishiji Biotechnology, Beijing, China). Then nitrocellulose membranes were washed 4 times using PBST, each time for 15 minutes. The GAPDH content was analyzed as the loading control with rabbit polyclonal antibody (Sigma, USA).

Liu Q., Cai J., Gao Y., Yang J., Gong Y, & Zhang Z. (2018). miR-2954 Inhibits PI3K Signaling and Induces Autophagy and Apoptosis in Myocardium Selenium Deficiency. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 51(2).

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