Phospho-PKC alpha (T638) 抗体 (YA163)

Phospho-PKC alpha (T638) Antibody is a non-conjugated and Rabbit origined monoclonal antibody about 77 kDa, targeting to Phospho-PKC alpha (T638). It can be used for WB,ICC/IF,IHC-P,IP assays with tag free, in the background of Human, Mouse, Rat.

PKC alpha 是一种钙激活、依赖磷脂和二酰基甘油 (DAG) 的丝氨酸/苏氨酸蛋白激酶，通过直接磷酸化 RAF1、BCL2、CSPG4、TNNT2/CTNT 等靶点或激活涉及 MAPK1/3 (ERK1/2) 和 RAP1GAP 的信号级联，参与细胞增殖、凋亡、分化、迁移、粘附、肿瘤发生、心肌肥大、血管生成、血小板功能和炎症的正负调控。它在神经胶质瘤细胞中通过磷酸化激活 RAF1 或上调 CDKN1A 促进细胞生长，同时在 PMA 刺激的肠细胞中触发细胞周期阻滞程序。PKC alpha 在巨噬细胞分化中易位至细胞核，参与伤口愈合时的粘附调节，并通过磷酸化 CSPG4 增强细胞运动性。在多种癌细胞中高表达，可作为肿瘤促进因子，并参与心肌收缩力的负调控及血管生成、血小板聚集的正调控。此外，它通过磷酸化心脏肌钙蛋白 T (TNNT2/CTNT) 调节心肌细胞功能，并在血管生成中调控内皮细胞迁移和 VEGFA 信号。在血小板中，它通过 CD36/GP4 受体和 RAP1GAP 通路激活 ITGA2B-ITGB3 整合素。PKC alpha 还参与 LPS 诱导的巨噬细胞功能调控，并通过磷酸化 EIF4G1、KIT、ATF2 等靶点影响翻译和转录。类似功能 (By similarity) 包括磷酸化 SOCS2 促进其降解，以及在血管紧张素 II 信号中磷酸化 KLHL3 以调节 WNK4 相互作用。最新研究发现 (PubMed:36040231)，它还磷酸化并激活 LRRK1，进而调控细胞内运输相关的 RAB 蛋白。

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: 1:500-1:5,000 ;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

液体

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.