Horseradish peroxidase hrp conjugated goat anti rabbit

Manufactured by Abcam

Sourced in United States

Horseradish peroxidase (HRP)-conjugated goat anti-rabbit is a secondary antibody that binds to rabbit primary antibodies. HRP is an enzyme that can be used to detect and quantify the presence of the target antigen in a sample.

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Lab products found in correlation

Pvdf membrane, by Merck Group (3 mentions) Ecl kit, by Thermo Fisher Scientific (2 mentions) Pierce ip lysis buffer, by Thermo Fisher Scientific (2 mentions) Pvdf membrane, by Abcam (1 mentions) Ecl kit, by Santa Cruz Biotechnology (1 mentions) Rabbit anti pten antibody, by Cell Signaling Technology (1 mentions) Chemiluminescent reagent, by Merck Group (1 mentions) Enhanced chemiluminescence, by Merck Group (1 mentions) Ripa lysis buffer, by Beyotime (1 mentions) Bca assay kit, by Beyotime (1 mentions) Ab203070, by Abcam (1 mentions) Ripa buffer, by Beyotime (1 mentions) Ab9485, by Abcam (1 mentions) Bicinchoninic acid protein assay kit, by Beyotime (1 mentions) Hrp dab chromogenic substrate kit, by Tiangen Biotech (1 mentions) Rabbit anti β tubulin antibody, by Abcam (1 mentions) Anti caspase 3, by Abcam (1 mentions) Anti cleaved caspase 3, by Abcam (1 mentions)

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5 protocols using horseradish peroxidase hrp conjugated goat anti rabbit

PTEN Protein Expression Analysis

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Total proteins were lysed by RIPA buffer (added with protease inhibitors) on ice and then were centrifuged at 12,000 × g at 4°C for 5 min. A mixture of equal protein and sample buffer was subjected to 15% SDS-PAGE. The separated protein was then transferred onto a PVDF membrane (Millipore Corporation, Billerica, MA, USA). The PVDF membrane was blocked in blocking buffer (containing TBST and 5% fat-free milk) for 1 h at room temperature. Primary antibody (rabbit anti-PTEN antibody, 1:1,000) was purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). GAPDH acted as the internal reference. PVDF membrane was incubated in the diluted primary antibodies at 4°C overnight. The PVDF membrane was then incubated with horseradish peroxidase (HRP)-conjugated goat anti-rabbit (1:500; Abcam, Cambridge, MA, USA) at 4°C overnight. Immunoblots were visualized by ECL kit (Santa Cruz Biotechnology, Santa Cruz, CA, USA) and scanned by ImageJ software. The band densities of each sample were normalized to the GAPDH band.

Qin N., Tong G.F., Sun L.W, & Xu X.L. (2017). Long Noncoding RNA MEG3 Suppresses Glioma Cell Proliferation, Migration, and Invasion by Acting as a Competing Endogenous RNA of miR-19a. Oncology Research, 25(9), 1471-1478.

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Detecting Donor HLA in Circulating Exosomes

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To detect donor HLA in circulating exosomes, we used the enzyme-linked immunosorbent assay (ELISA) method as described by Logozzi M et al, [17 (link)] with minor modifications. We set up an ELISA protocol for measuring HLA-A2 in the circulating exosomes since HLA-A2 is one of the high frequency HLA antigen in our population. In brief, antibodies (Abs) to CD9 (Bio-Legend) were coated in 96-well plates, followed by blocking with 0.5% bovine serum albumin in PBS) Circulating exosomes (1mg/ml) isolated from sera of LTxRs were then added and incubated at 37°C for 12 hours, followed by addition of a rabbit Ab specific to HLA-A2 (Abcam) and secondary Ab, horseradish peroxidase (HRP) conjugated goat anti-rabbit (Abcam). Reaction was developed with chemiluminescent reagent (Millipore) for 15 minutes and stopped with 0.1N HCl. Optical density was measured at 450 nm. Data were expressed as mean ± standard deviation, and p-values were deemed significant at less than 0.05.

Sharma M., Ravichandran R., Perincheri S., Danziger-Isakov L., Heeger P.S., Sweet S.C, & Mohanakumar T. (2020). Distinct Molecular and Immunological Properties of Circulating exosomes Isolated from Pediatric Lung Transplant Recipients with Bronchiolitis Obliterans Syndrome. Transplant international : official journal of the European Society for Organ Transplantation, 33(11), 1491-1502.

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Western Blot Protein Analysis Protocol

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Proteins from cells were obtained in protein extraction reagent RIPA lysis buffer (Beyotime) with 10 mM phenylmethylsulphonyl fluoride (Beyotime) at 4 °C for 30 min. The supernatant was harvested after centrifugation at 12000 rpm for 30 min. A BCA assay kit (Beyotime) was used to determine protein concentration. Then, equal amounts of protein samples were added to the loading buffer (Beyotime). After heating at 95 °C for 5 min, each sample was run on a 10% SDS polyacrylamide gel. The separated proteins were electrically transferred onto PVDF membrane (Millipore,USA), which were then blocked with 5% (w/v) no-fat dry milk in TBST for 1 h at room temperature. After blocking, the membranes were incubated with primary antibodies (Supplement Table 3) overnight at 4 °C, followed by incubation in horseradish peroxidase (HRP)-conjugated goat anti-rabbit (Abcam, UK) for 1 h. Immunoreactive bands were visualized by enhanced chemiluminescence (Millipore) and quantitative analyzed by Image J software. β-actin was used as an internal control standard to analyze relative protein expression.

JIANG M., SHEN Q., ZHOU Y., REN W., CHAI M., ZHOU Y, & TAN W.S. (2021). Fluid shear stress and endothelial cells synergistically promote osteogenesis of mesenchymal stem cells via integrin β1-FAK-ERK1/2 pathway. Turkish Journal of Biology, 45(6), 683-694.

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BAMBI Protein Extraction and Detection

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Total protein extraction was accomplished using RIPA buffer (Beyotime, Shanghai, China). After measurement of the protein concentration using a bicinchoninic acid protein assay kit (Beyotime), the protein samples were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) separation and transferred onto a polyvinylidene difluoride (PVDF) membrane. The membrane was blocked with 5% skim milk and incubated with specific primary antibodies against BAMBI (ab203070; 1:1,000, Abcam, Cambridge, MA, USA) and GAPDH (ab9485; 1:2000, Abcam, Cambridge, MA, USA) at 4 °C. After incubation overnight, the membrane was incubated with horseradish peroxidase (HRP)-conjugated goat anti-rabbit (1:2,000, Abcam, Cambridge, MA, USA) secondary antibody for 2 hours at room temperature. Finally, the signals were detected using an electrochemiluminescence (ECL) kit (Thermo Fisher Scientific, Inc.). GAPDH was used as the internal control.

Pan Z., Mo F., Liu H., Zeng J., Huang K., Huang S., Cao Z., Xu X., Xu J., Liu T, & Huang J. (2021). LncRNA prostate androgen-regulated transcript 1 (PART 1) functions as an oncogene in osteosarcoma via sponging miR-20b-5p to upregulate BAMBI. Annals of Translational Medicine, 9(6), 488.

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DUSP5 Overexpression Modulates SGIV Infection

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GS cells transfected with 800 ng pcDNA-3.1-3HA/pcDNA-3.1-DUSP5 for 24 h were infected with SGIV for 24 h. The cells were collected and lysed by pierce IP lysis buffer (Thermo Fisher), then separated by 10% sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE). Subsequently, the proteins were transferred onto 0.2 μm PVDF membranes (Millipore, Billerica, MA, USA) and blocked using 5% skim milk for 2 h. The membranes were incubated with rabbit anti-MCP antibody (1:1000 dilution), which was prepared in our lab with the patent number CN111363758A [19 (link)]. Rabbit anti-β-tubulin antibody (1:2000 dilution), anti-3HA antibody (1:1000 dilution), anti-caspase3 (1:1000 dilution), and anti-cleaved caspase3 (1:1000 dilution) were purchased from Abcam. The horseradish peroxidase (HRP)-conjugated goat anti-rabbit (1:5000 dilution) was purchased from Abcam as well. Immuno-reactive proteins were visualized by an enhanced HRP-DAB Chromogenic Substrate Kit (Tiangen, Beijing, China).

He J., Cai Y., Huang W., Lin Y., Lei Y., Huang C., Cui Z., Qin Q, & Sun H. (2023). The Role of Epinephelus coioides DUSP5 in Regulating Singapore Grouper Iridovirus Infection. Viruses, 15(9), 1807.

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