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

Applications

IP-WB, WB, ELISA

Conjugate

HRP

Host

Goat

Reactivity

Rabbit

Cat no : RGAR301

Validation Data Gallery

  • 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.

    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.

  • 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.

    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.

  • 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.

    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.

  • 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.

    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.

  • 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.

    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.

  • 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.

    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.

  • 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.

    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.

  • 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.

    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.

  • 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.

    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.

  • 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.

    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.

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.
ClonalityRecombinant monoclonal
RRIDAB_3712952

背景介绍

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.
Chinese Version:
本产品特异性识别天然兔IgG分子，但与完全还原的兔IgG片段（包括重链和轻链）无交叉反应。这一特性使其成为免疫共沉淀-蛋白印迹（IP-WB）检测的理想工具，因为IP过程中使用的一抗在样品制备时会被还原成片段，在后续的Western blot检测中，一抗片段可被常规二抗识别（重链约55 kDa，轻链约25 kDa），而使用本构象特异性二抗时，还原片段则不会被检测到。当目标蛋白分子量接近55 kDa或25 kDa时尤为重要，使用本二抗可以避免由内源信号带来的误判。