PKCα Antibody [B21A18] | Primary Antibodies

Application: Reactivity: Human, Mouse, Rat

Lane 1: SH-SY5Y, Lane 2: NIH/3T3

Usage Information

Dilution
WB1:1000 IF1:100 FCM1:200

Application
WB, IF, FCM

Reactivity
Human, Mouse, Rat

Source
Rabbit Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW
80 kDa

Positive Control	SH-SY5Y cells; NIH/3T3 cells; SK-N-SH cells (TPA, 200 nM, 30 min)
Negative Control	SK-MEL-2 cells

Datasheet & SDS

Biological Description

Specificity
PKCα Antibody [B21A18] detects endogenous levels of total PKCα protein.

Clone
B21A18

Synonym(s)
PRKCA; Protein kinase C alpha type; EC:2.7.11.13; PKC-A; PKC-alpha; PKCA; PRKACA

Background
PKCα (protein kinase C alpha) is a classical, calcium-dependent serine/threonine kinase that plays a pivotal role in transducing signals for diverse cellular responses, including gene expression, secretion, proliferation, and muscle contraction. As a member of the PKC family, PKCα is activated by the coordinated binding of calcium ions to its C2 domain and phosphatidylserine and diacylglycerol (DAG) to its cysteine-rich C1 domain, with phorbol esters serving as potent pharmacological activators. The protein features an autoinhibitory pseudosubstrate domain that occupies the substrate-binding pocket of the catalytic domain, maintaining PKCα in an inactive conformation until cofactor or activator binding induces a conformational change. Full activation requires a series of phosphorylation events, notably at threonine and serine residues within the activation loop (Thr500), turn motif (Thr641), and hydrophobic motif (Ser660), often mediated by PDK1 and autophosphorylation, which stabilize the active conformation and promote downstream signaling. PKCα translocates to the plasma membrane upon activation, where it phosphorylates a wide spectrum of substrates involved in cytoskeletal remodeling, ion channel regulation, and transcriptional control, thus integrating extracellular cues into precise cellular outcomes. Dysregulation of PKCα activity has been implicated in cardiovascular disease, cancer progression, and neurological disorders.

Background

PKCα (protein kinase C alpha) is a classical, calcium-dependent serine/threonine kinase that plays a pivotal role in transducing signals for diverse cellular responses, including gene expression, secretion, proliferation, and muscle contraction. As a member of the PKC family, PKCα is activated by the coordinated binding of calcium ions to its C2 domain and phosphatidylserine and diacylglycerol (DAG) to its cysteine-rich C1 domain, with phorbol esters serving as potent pharmacological activators. The protein features an autoinhibitory pseudosubstrate domain that occupies the substrate-binding pocket of the catalytic domain, maintaining PKCα in an inactive conformation until cofactor or activator binding induces a conformational change. Full activation requires a series of phosphorylation events, notably at threonine and serine residues within the activation loop (Thr500), turn motif (Thr641), and hydrophobic motif (Ser660), often mediated by PDK1 and autophosphorylation, which stabilize the active conformation and promote downstream signaling. PKCα translocates to the plasma membrane upon activation, where it phosphorylates a wide spectrum of substrates involved in cytoskeletal remodeling, ion channel regulation, and transcriptional control, thus integrating extracellular cues into precise cellular outcomes. Dysregulation of PKCα activity has been implicated in cardiovascular disease, cancer progression, and neurological disorders.

References
https://pubmed.ncbi.nlm.nih.gov/10928943/ https://pubmed.ncbi.nlm.nih.gov/35492590/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%