Immunohistochemistry for Ki67 and Vimentin was performed on the 4% paraformaldehyde-fixed, paraffin-embedded subcutaneous tumor tissues using a primary antibody against target antigen (anti-Ki67, Abcam, USA; anti-Vimentin, Cell Signaling Technology, Inc) and a horseradish peroxidase-conjugated IgG (Abcam, USA). The proteins in situ were visualized with 3,3-diaminobenzidine.
Horseradish peroxidase conjugated igg
Manufactured by Abcam
Sourced in United States
Horseradish peroxidase-conjugated IgG is a secondary antibody conjugated with the enzyme horseradish peroxidase. The core function of this product is to detect and quantify target proteins in various immunoassay techniques, such as Western blotting, ELISA, and immunohistochemistry.
Automatically generated - may contain errors
Lab products found in correlation
Anti ki67, by Abcam (1 mentions)
Anti vimentin, by Cell Signaling Technology (1 mentions)
Immobilon p pvdf membrane, by Merck Group (1 mentions)
Anti myb, by Abcam (1 mentions)
Anti mdr1, by Abcam (1 mentions)
Anti mrp1, by Abcam (1 mentions)
Complete protease inhibitor cocktail, by Merck Group (1 mentions)
Anti β actin, by Abcam (1 mentions)
Nuclear protein extraction kit, by Thermo Fisher Scientific (1 mentions)
Ecl detection kit, by Thermo Fisher Scientific (1 mentions)
Histone h3, by Abcam (1 mentions)
Polyvinylidene fluoride membrane, by Merck Group (1 mentions)
Mcp 1, by Abcam (1 mentions)
Chemidoc mp system, by Bio-Rad (1 mentions)
Gapdh, by Abcam (1 mentions)
4 protocols using horseradish peroxidase conjugated igg
1
Immunohistochemical Analysis of Tumor Markers
2
Protein Expression Analysis of MDR Factors
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Total cellular protein was extracted using ice-cold RIPA lysis buffer (50 mM Tris–HCl, pH = 7.4, 150 mM NaCl, 0.1% SDS, 1% Triton X-100, 0.5% sodium deoxycholate) containing 1× complete protease inhibitor cocktail (EDTA-free, Sigma-Aldrich). Protein extracts were electrophoresed on a 10% SDS polyacrylamide gel and transferred to the Immobilon-P PVDF membrane (Millipore, Billerica, MA, USA). The following primary antibodies were used: anti-multidrug resistance-related protein 1 (anti-MRP1, Abcam, Cambridge, UK; dilution 1 : 1000), anti-multidrug resistance 1 (anti-MDR1, Abcam; dilution 1 : 2000), anti-Myb (Abcam; dilution 1 : 1000) and anti-β-actin (Abcam; dilution 1 : 5000). Horseradish peroxidase-conjugated IgG (Abcam; dilution 1 : 5000) was used a second antibody. Protein bands were detected using ECL reagent (GE Healthcare Technologies, Waukesha, WI, USA) and analyzed by ImageJ software (National Institutes of Health, Bethesda, MD, USA).
3
Nuclear Protein Extraction and Western Blot
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Nuclear protein was extracted from lymphocytes using the Nuclear Protein Extraction Kit (Thermo Scientific). Protein concentrations were determined using Bradford method. Equal amounts were separated on an sodium dodecyl sulfate‐polyacrylamide gel electrophoresis gel, and transferred onto polyvinylidene fluoride membranes at 4°C. The membranes were subsequently blocked and incubated with primary antibodies against PAX6 (1:1,000; Abcam) and Histone H3 (1:3,000; Abcam) overnight at 4°C. Horseradish peroxidase‐conjugated IgG (1:5,000; Abcam) was used as a secondary antibody for incubation at room temperature for 2 hr. The final detection reaction was performed with the ECL detection kit (Thermo Scientific).
4
Western Blot Analysis of Inflammatory Proteins
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Antibodies against the following proteins were used for Western blot analysis: caspase 1 and caspase 4 (human), caspase 11 (mouse), GSDMD, MCP1, CCR2 (1:1000, Abcam), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH; 1:10000, Abcam). The secondary antibody was horseradish peroxidase-conjugated IgG (1:8000, Abcam). For the blots, 30–50 µg of total protein was added into each well and the proteins were separated by electrophoresis on a 10% sodium dodecylsulphate polyacrylamide gel electrophoresis precast gel (Invitrogen, CA, United States). The proteins were then transferred to polyvinylidene fluoride membranes (Millipore, Bedford, MA, United States). The protein bands were developed with an enhanced luminescent developer (Millipore) and photographed using a Chemi Doc MP System (Bio-Rad, Hercules, United States). The grey value was measured using ImageJ software.
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