Phospho-PKC alpha (Thr638) Polyclonal Antibody

$Image kindly provided by Dr. Magdalena Krol. Control tumor cells, tumor cells grown in macrophage-conditioned medium, tumor cells sorted from co-culture with macrophages, and macrophages from monocultures and sorted from co-culture with tumor cells were analyzed. Total protein concentrations in lysates were determined using a Bio-Rad protein assay. Proteins (50 mg) were resolved using SDS-PAGE and transferred onto PVDF membranes. The membranes were then blocked with 5% non-fat dry milk in TBS buffer containing 0.5% Tween 20. The membranes were then incubated overnight with the primary Rabbit Anti-PKC alpha\/beta II (Thr638\/641) Polyclonal Antibody at 1:100 dilution. Subsequently, the membranes were washed three times in TBS containing 0.5% Tween 20 and incubated for 1 h at room temperature with secondary antibodies conjugated with the appropriate infrared (IR) fluorophore IRDyeH 800 CW or IRDyeH 680 RD at a dilution of 1:5000.$

Image kindly provided by Dr. Magdalena Krol. Control tumor cells, tumor cells grown in macrophage-conditioned medium, tumor cells sorted from co-culture with macrophages, and macrophages from monocultures and sorted from co-culture with tumor cells were analyzed. Total protein concentrations in lysates were determined using a Bio-Rad protein assay. Proteins (50 mg) were resolved using SDS-PAGE and transferred onto PVDF membranes. The membranes were then blocked with 5% non-fat dry milk in TBS buffer containing 0.5% Tween 20. The membranes were then incubated overnight with the primary Rabbit Anti-PKC alpha\/beta II (Thr638\/641) Polyclonal Antibody at 1:100 dilution. Subsequently, the membranes were washed three times in TBS containing 0.5% Tween 20 and incubated for 1 h at room temperature with secondary antibodies conjugated with the appropriate infrared (IR) fluorophore IRDyeH 800 CW or IRDyeH 680 RD at a dilution of 1:5000.

$Formalin-fixed and paraffin embedded mouse brain labeled with Rabbit Anti-PKC alpha\/beta II (Thr638\/641) Polyclonal Antibody, Unconjugated (bs-3333R) at 1:200 followed by conjugation to the secondary antibody and DAB staining\\n$

Formalin-fixed and paraffin embedded mouse brain labeled with Rabbit Anti-PKC alpha\/beta II (Thr638\/641) Polyclonal Antibody, Unconjugated (bs-3333R) at 1:200 followed by conjugation to the secondary antibody and DAB staining\\n

Applications

WB
ELISA
IHC-P
IHC-F
IF(IHC-P)
IF(IHC-F)
IF(ICC)

Reactivity

Human
Mouse
Rat

Predicted Reactivity

Dog
Cow
Horse
Chicken

Overview
Catalog #	bs-3333R
Product Name	Phospho-PKC alpha (Thr638) Polyclonal Antibody
Applications	WB ELISA IHC-P IHC-F IF(IHC-P) IF(IHC-F) IF(ICC)
Reactivity	Human, Mouse, Rat
Predicted Reactivity	Dog, Cow, Horse, Chicken
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	KLH conjugated synthetic phosphopeptide derived from human PKC alpha/beta II around the phosphorylation site of Thr638/641
Modification Site	Thr638/641
Clonality	Polyclonal
Isotype	IgG
Concentration	1ug/ul
Purification	Purified by Protein A.
Storage Buffer	0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage Condition	Shipped at 4_. Store at -20_ for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID	5578
Swiss Prot	P17252
Subcellular location	Cytoplasm, Nucleus, Cell membrane
Synonyms	AAG6; PKCA; PRKACA; PKC-alpha; Protein kinase C alpha type; PKC-A; PRKCA
Background	Protein Kinase c alpha (PKC alpha) is an 77 kDa member of the conventional group (cPKCs: sensitive to calcium, diacylglycerol, phosphatidylserine and phorbol esters) of the PKC family of serine/ threonine kinases that are involved in a wide range of physiological processes including mitogenesis, cell survival and transcriptional regulation. PKC alpha is an ubiquitously expressed PKC isozyme that has been implicated in the regulation of a broad range of cellular functions including proliferation, differentiation, development, migration, cell cell adhesion, cell extracellular matrix adhesion, and solute transport. The activation loop threonine (threonine 497 in PKC alpha) of conventional PKCs is phosphorylated by phosphoinositide dependent kinase 1 (PDK1). This phosphorylation is necessary for the autophosphorylation of threonine 638 in the carboxy terminus of PKC alpha, a step that is critical for regulating the rate of PKC alpha dephosphorylation and inactivation.
Application Dilution
WB	1:300-5000
ELISA	1:500-1000
IHC-P	1:200-400
IHC-F	1:100-500
IF(IHC-P)	1:50-200
IF(IHC-F)	1:50-200
IF(ICC)	1:50-200