HRP-Goat Anti-Rabbit IgG Conformation Specific Recombinant Secondary Antibody

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  IP-WB detection with RGAR301. Lysate: HeLa cells; IP Primary: RPLP0, 11290-2-AP; IP Control: Rabbit IgG, 30000-1-AP; SDS-PAGE loading amount: 20% of IP elution and 30 μg total protein for input; WB Primary: RPLP0, 11290-2-AP, 1:8000; WB secondary: RGAR301, 1:2500.

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  IP-WB detection with RGAR301. Lysate: HEK-293 cells; IP Primary: TEX264, 25858-1-AP; IP Control: Rabbit IgG, 30000-1-AP; SDS-PAGE loading amount: 20% of IP elution and 30 μg total protein for input; WB Primary: TEX264, 25858-1-AP, 1:3000; WB secondary: RGAR301, 1:2500.

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  IP-WB detection with RGAR301. Lysate: HepG2 cells; IP Primary: TMCO3, 31488-1-AP; IP Control: Rabbit IgG, 30000-1-AP; SDS-PAGE loading amount: 20% of IP elution and 30 μg total protein for input; WB Primary: TMCO3, 31488-1-AP, 1:1000; WB secondary: RGAR301, 1:2500.

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  IP-WB detection with RGAR301. Lysate: Raji cells; IP Primary: AIM2, 25090-1-AP; IP Control: Rabbit IgG, 30000-1-AP; SDS-PAGE loading amount: 20% of IP elution and 30 μg total protein for input; WB Primary: AIM2, 25090-1-AP, 1:600; WB secondary: RGAR301, 1:2500.

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  IP-WB detection with RGAR301. Lysate: HepG2 cells; IP Primary: Annexin A2, 11256-1-AP; IP Control: Rabbit IgG, 30000-1-AP; SDS-PAGE loading amount: 20% of IP elution and 30 μg total protein for input; WB Primary: Annexin A2, 11256-1-AP, 1:30000; WB secondary: RGAR301, 1:2500.

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  IP-WB detection with RGAR301. Lysate: Mouse brain tissue; IP Primary: Synaptophysin, 17785-1-AP; IP Control: Rabbit IgG, 30000-1-AP; SDS-PAGE loading amount: 20% of IP elution and 30 μg total protein for input; WB Primary: Synaptophysin, 17785-1-AP, 1:50000; WB secondary: RGAR301, 1:2500.

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  Rabbit recombinant antibodies (84975-1-RR and 84871-1-RR) and rabbit polyclonal antibodies (15240-1-AP and 15675-1-AP) were subjected to SDS-PAGE at reduced condition at 100 ng/well followed by western blotting with Multi-rAb™ HRP-Goat Anti-Rabbit Recombinant Secondary Antibody (H+L) RGAR001 (0.05 μg/mL) and HRP-Goat Anti-Rabbit IgG Conformation Specific Recombinant Secondary Antibody RGAR301 (0.1 μg/mL). The result suggest that RGAR301 does not react with reduced fragment of rabbit IgGs.

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  Various lysates were subjected to SDS PAGE followed by western blot with 15282-1-AP (HSP60 antibody) at dilution of 1:100000. HRP-Goat Anti-Rabbit IgG Conformation Specific Recombinant Secondary Antibody #RGAR301 was used at 1:5000 for detection.

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  Western blot detection using RGAR301. Various lysates were subjected to SDS PAGE followed by western blot with 15282-1-AP (HSP60 antibody) at dilution of 1:100000. HRP-Goat Anti-Rabbit IgG Conformation Specific Recombinant Secondary Antibody #RGAR301 was used at 1:10000 for detection.

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  Rabbit IgG was coated at 50 ng/well followed by detection with serial diluted secondaries. Signal was developed with TMB and stopped by H2SO4. Signal strength was measured by absorbance at 450 nm. Blue curve: Proteintech's Multi-rAb™ HRP-Goat Anti-Rabbit Recombinant Secondary Antibody (H+L) #RGAR001; Green curve: Proteintech's HRP-Goat Anti-Rabbit IgG Conformation Specific Recombinant Secondary Antibody #RGAR301; Orange curve: Competitor's Mouse Anti-rabbit IgG Conformation Specific secondary antibody.

Cat No. RGAR301

Applications

IP-WB, WB, ELISA

Conjugate

HRP

Host

Goat

Reactivity

Rabbit

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Product Information

• Concentration: 0.5 mg/mL
• Applications: IP-WB, WB, ELISA
• Host Species: Goat
• Species Reactivity: Rabbit
• Physical State: Liquid
• Suggested Dilution Range: 1:3000-1:10,000 for ELISA;; 1:2500-1:5000 for western blotting or IP-WB with ECL substrates.
• Purity: The antibody was purified from culture media supernatant by immunoaffinity chromatography using Protein G beads.
• Storage Buffer: PBS with 50% glycerol, 10 mg/mL BSA, 0.1% Proclin-300, pH 7.4.
• Storage Instructions: Store at -20°C. Stable for one year after shipment.
• Clonality: Recombinant monoclonal
• RRID: AB_3712952

Background

This HRP-Goat Anti-Rabbit IgG Conformation Specific Recombinant Secondary Antibody (RGAR301) strongly reacts with native rabbit IgG and does not react with fully reduced rabbit IgG fragments (heavy chain and light chain). It is a very useful tool in IP-WB detection because the primary antibody that used for immunoprecipitation is reduced during sample preparation and could be detected with regular secondary antibody at ~55 kDa (heavy chain) and ~25 kDa (light chain) in the subsequent western blot, while the fragments would not be detected using this secondary. This is especially beneficial if the molecular weight of target protein is close to ~55 kDa or ~25 kDa as there won't be confusion about the signal source when using this product.