LS-C60073

antibody from LSBio

Targeting: CREB1

ELISA

Immunocytochemistry

Immunohistochemistry

Antibody data

Submit

Validation data

CREB1 protein structure - LS-C60073 shown in red.

Supportive validation

Submitted by: LSBio (provider)
Enhanced method: Genetic validation
Main image
Experimental details: Anti-CREB pS133 Antibody - Western Blot. Anti-CREB pS133 was used to detect phosphorylated CREB by western blot. Recombinant His-tagged human CREB was produced in E. coli and purified by metal affinity chromatography. An aliquot of purified CREB was phosphorylated in-vitro using Protein Kinase A and ATP. Western blot of indicated amounts of control (-) and in-vitro phosphorylated CREB (P) were loaded to show that the antibody reacts specifically with the phosphorylated form. Blots were blocked in 5% milk in TBS+0.1% Tween-20 (TBST-M) overnight at 4C. Detection occurs using a 1:500 dilution of antibody diluted in TBST-M and incubated at room temperature with rocking for 1 hour. Blots were rinsed 6X with TBST and incubated with goat anti-rabbit-HRP at 1:5000 in TBST-M at room temperature for 45 min. Blots were again rinsed 6X with TBST and then processed using ECL reagent (Amersham) according to manufacturers instructions. Exposure time: 1 min using Kodak Biomax MR film. Personal Communication, R. Screaton, The Salk Institute for Biological Studies.

Supportive validation

Submitted by: LSBio (provider)
Enhanced method: Genetic validation
Main image
Experimental details: Anti-CREB pS133 Antibody - Western Blot. Anti-CREB pS133 was used to detect phosphorylated CREB by western blot. Recombinant His-tagged human CREB was produced in E. coli and purified by metal affinity chromatography. An aliquot of purified CREB was phosphorylated in-vitro using Protein Kinase A and ATP. western blot of control (-) and in-vitro phosphorylated CREB (+) was used to show that the antibody reacts specifically with the phosphorylated form. Pan reactive CREB (# LS-C19089) reacts equally with both non-phosphorylated and phosphorylated CREB (not shown). Detection occurs using a 1:500 dilution of antibody followed by 1:5000 dilution of HRP Goat-a-Rabbit IgG with visualization via ECL. Film exposure was approximately 1. Other detection systems will yield similar results. Personal Communication, Boss, J., Emory University School of Medicine, Atlanta, GA.