Phospho-PKC alpha (T638) Antibody (YA163)

Phospho-PKC alpha (T638) Antibody (YA163) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to Phospho-PKC alpha (T638).

PKC alpha is a calcium-activated, phospholipid- and diacylglycerol (DAG)-dependent serine/threonine-protein kinase that regulates cell proliferation, apoptosis, differentiation, migration, adhesion, tumorigenesis, cardiac hypertrophy, angiogenesis, platelet function, and inflammation by directly phosphorylating targets like RAF1, BCL2, CSPG4, and TNNT2/CTNT or activating signaling cascades involving MAPK1/3 (ERK1/2) and RAP1GAP. It promotes cell growth in glioma cells via RAF1 phosphorylation or CDKN1A upregulation, while inducing cell cycle arrest in PMA-stimulated intestinal cells. During macrophage differentiation, it translocates to the nucleus and modulates desmosomal adhesion post-wounding. PKC alpha enhances cell motility by phosphorylating CSPG4 and is overexpressed in cancers, acting as an oncogene. It negatively regulates myocardial contractility but positively controls angiogenesis and platelet aggregation. The kinase phosphorylates cardiac troponin T (TNNT2/CTNT) to reduce myocardial contractility and stabilizes VEGFA mRNA to promote angiogenesis. In platelets, it mediates granule release via CD36/GP4 and activates ITGA2B-ITGB3 integrin through RAP1GAP. PKC alpha also modulates LPS-induced macrophage functions and negatively regulates NF-κB via IL1A-dependent NFKBIA induction. It phosphorylates EIF4G1, KIT, and ATF2 to influence translation and transcription. Similar functions (By similarity) include SOCS2 phosphorylation for degradation and KLHL3 phosphorylation to disrupt WNK4 binding. Recent studies (PubMed:36040231) show it activates LRRK1 to regulate RAB proteins in intracellular trafficking.

Free

P17252

Predicted band size: 77 kDa; Observed band size: 77 kDa

Protein A affinity purified.

Cytoplasm, Cell membrane, Mitochondrion membrane, Nucleus.

Non-conjugated

Phosphorylated

AB_3102400

Signal Transduction

Primary Antibody; Recombinant Rabbit Monoclonal Antibody

Recombinant, Monoclonal

Rabbit

Human, Mouse, Rat

WB: 1:500-1:5000; ICC/IF: 1:50-1:500; IHC-P: 1:50-1:500; IP: Use at an assay dependent concentration.

• 
  Western blot analysis of extracts from NIH3T3 (lane 2(20μg)，C6 (lane 3(20μg), Hela (lane 4(20μg) and HEK293(lane 5(20μg) using Phospho-PKC alpha (T638) (HY-P80279) 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 IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature.
• 
  Immunocytochemistry analysis of Hela cells labeling Phospho-PKC alpha (T638) with Phospho-PKC alpha (T638) Antibody (HY-P80279)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-PKC alpha (T638) Antibody (HY-P80279) 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 brain tissue using Phospho-PKC alpha (T638) Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0) 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 (HY-P80279, 1/500) 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 brain tissue using Phospho-PKC alpha (T638) Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0) 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 (HY-P80279, 1/500) 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

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 Thr638 of Human PKC alpha.AA range:622-667.