Phospho-CREB (Ser133) Antibody (YA210)

Phospho-CREB (Ser133) Antibody (YA210) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to Phospho-CREB (Ser133).

Creb is a phosphorylation-dependent transcription factor that stimulates transcription by binding to the cAMP response element (CRE) in DNA, a sequence found in many viral and cellular promoters (By similarity). Its transcriptional activation is enhanced by TORC coactivators, which function independently of Ser-119 phosphorylation (PubMed:14536081). Creb is involved in various cellular processes, including the synchronization of circadian rhythms and the differentiation of adipose cells (By similarity). Additionally, it regulates the expression of apoptotic and inflammatory response factors in cardiomyocytes in response to ERFE-mediated activation of AKT signaling (By similarity).

Free

P16220

Predicted band size: 35 kDa; Observed band size: 42 kDa

Protein A affinity purified.

Nucleus.

Non-conjugated

Phosphorylated

AB_3102531

Epigenetics and Nuclear Signaling

Primary Antibody; Recombinant Rabbit Monoclonal Antibody

Recombinant, Monoclonal

Rabbit

Human, Mouse

WB: 1:500-1:1,000 ;ICC: 1:50-1:100 ;IHC-P: 1:50-1:400 ;FC: 1:50-1:100 ;IP: Use at an assay dependent concentration.

• 
  Western blot analysis of extracts from HEK293(lane 2(20μg) using Phospho-Creb(HY-P80457 Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature.
• 
  Immunocytochemistry analysis of C6 cells labeling Phospho-Creb (Ser133) with Phospho-Creb (Ser133) Antibody (HY-P80457) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Phospho-Creb (Ser133) Antibody (HY-P80457) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunocytochemistry analysis of NIH3T3 cells labeling Phospho-Creb (Ser133) with Phospho-Creb (Ser133) Antibody (HY-P80457) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Phospho-Creb (Ser133) Antibody (HY-P80457)at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002,Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunohistochemical analysis of paraffin-embedded rat colon tissue using Phospho-Creb Antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/100 dilution in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
• 
  Immunohistochemical analysis of paraffin-embedded rat colon tissue using Phospho-Creb Antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/100 dilution in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

WB, ICC/IF, IHC-P, IP, FC

Liquid

Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping with blue ice.

IgG

Endogenous

Synthetic phosphopeptide corresponding to residues surrounding Ser133 of Human Creb.The exact sequence is proprietary to MCE.