Immunoprecipitation of PRAS40 from HeLa cells using PRAS40 (D23C7) XP® Rabbit mAb #2691 (lane 2). Lane 1 contains input control lysate. All lanes are probed with PRAS40 (D23C7) Rabbit mAb #2691 as the primary antibody. Secondary antibodies include Anti-rabbit IgG, HRP-linked Antibody #7074 (panel A) and Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 (panel B).
Immunoprecipitation of Beclin-1 from HeLa cells using Beclin-1 (D40C5) Rabbit mAb #3495 (lane 2). Lane 1 contains input control lysate. All lanes are probed with Beclin-1 (D40C5) Rabbit mAb #3495 as the primary antibody. Secondary antibodies include Anti-rabbit IgG, HRP-linked Antibody #7074 (panel A) and Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 (panel B).
Affinity purified mouse anti-rabbit IgG (Conformation Specific) HRP Conjugate antibody.
Recommended Antibody Dilution:
Supplied in 136 mM NaCl, 2.6 mM KCl, 12 mM sodium phosphate (pH 7.4) dibasic, 2 mg/ml BSA, and 50% glycerol. Store at –20°C. Do not aliquot the antibodies.
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) recognizes the native rabbit IgG. This antibody may cross react with denatured [light chain] IgG at high concentrations.
Monoclonal antibody is produced by immunizing animals with native total rabbit IgG.
Mouse Anti-Rabbit IgG (Conformation Specific) (L27A9) mAb only reacts with native IgG and does not bind to the denatured and reduced rabbit IgG heavy chain or light chain. When performing immunopreciptiation (IP) followed by western blotting, the denatured rabbit IgG light and heavy chains of the primary antibody used for IP run at approximately 25 and 50 kD, respectively, on the subsequent western blot and can often obscure bands of proteins that have similar molecular weights. Using Mouse Anti-Rabbit IgG (Conformation Specific) (L27A9) mAb as a secondary antibody will eliminate this problem.
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