Phospho-PKC alpha (T497) Recombinant Rabbit Monoclonal Antibody [JE44-81]
Catalog# HA722278
Phospho-PKC alpha (T497) Recombinant Rabbit Monoclonal Antibody [JE44-81]
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WB
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IF-Cell
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FC
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
Phospho-PKC alpha (T497) Recombinant Rabbit Monoclonal Antibody [JE44-81]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic phosphopeptide corresponding to residues surrounding Thr497 of Human PKC alpha.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, FC
Molecular Weight
Predicted band size: 77 kDa
Positive Control
HeLa cell lysate, NIH/3T3 cell lysate, C6 cell lysate, MCF7 cells treated with 100nM PMA for 1 hour, HeLa, NIH/3T3, C6, K-562.
Conjugation
unconjugated
Clone Number
JE44-81
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
-
WB
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1:2,000
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IF-Cell
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1:100-1:3,000
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FC
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1:1,000
Target
Function
Protein kinase C alpha (PKCα) is an enzyme that in humans is encoded by the PRKCA gene. Protein kinase C (PKC) is a family of serine- and threonine-specific protein kinases that can be activated by calcium and the second messenger diacylglycerol. PKC family members phosphorylate a wide variety of protein targets and are known to be involved in diverse cellular signaling pathways. PKC family members also serve as major receptors for phorbol esters, a class of tumor promoters. Each member of the PKC family has a specific expression profile and is believed to play a distinct role in cells. The protein encoded by this gene is one of the PKC family members. This kinase has been reported to play roles in many different cellular processes, such as cell adhesion, cell transformation, cell cycle checkpoint, and cell volume control. Knockout studies in mice suggest that this kinase may be a fundamental regulator of cardiac contractility and Ca2+ handling in myocytes. Protein kinase C-alpha (PKC-α) is a specific member of the protein kinase family. These enzymes are characterized by their ability to add a phosphate group to other proteins, thus changing their function. PKC-α has been widely studied in the tissues of many organisms including drosophila, xenopus, cow, dog, chicken, human, monkey, mouse, pig, and rabbit. Many studies are currently being conducted investigating the structure, function, and regulation of this enzyme. The most recent investigations concerning this enzyme include its general regulation, hepatic function, and cardiac function.
Background References
1. Ghashghaeinia M et al. Coronavirus disease 2019 (COVID-19), human erythrocytes and the PKC-alpha/-beta inhibitor chelerythrine -possible therapeutic implication. Cell Cycle. 2020 Dec
2. Ghashghaeinia M et al. The specific PKC-alpha inhibitor chelerythrine blunts costunolide-induced eryptosis. Apoptosis. 2020 Oct
Subcellular Location
Cytoplasm, Cell membrane, Mitochondrion membrane, Nucleus.
Synonyms
AAG6 antibody
Aging associated gene 6 antibody
aPKC antibody
KPCA_HUMAN antibody
PKC alpha antibody
PKC-A antibody
PKC-alpha antibody
PKCA antibody
PRKACA antibody
PRKCA antibody
ExpandImages
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Western blot analysis of Phospho-PKC alpha (T497) on different lysates with Rabbit anti-Phospho-PKC alpha (T497) antibody (HA722278) at 1/2,000 dilution.
Lane 1: HeLa cell lysate
Lane 2: NIH/3T3 cell lysate
Lane 3: C6 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 77 kDa
Observed band size: 77 kDa
Exposure time: 1 minute; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722278) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Cell treatment (CT)
Immunocytochemistry analysis of MCF7 cells treated with 100nM PMA for 1 hour labeling Phospho-PKC alpha (T497) with Rabbit anti-Phospho-PKC alpha (T497) antibody (HA722278) at 1/3,000 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-PKC alpha (T497) antibody (HA722278) at 1/3,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
☑ Cell treatment (CT)
Immunocytochemistry analysis of HeLa cells treated with or without λpp labeling Phospho-PKC alpha (T497) with Rabbit anti-Phospho-PKC alpha (T497) antibody (HA722278) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-PKC alpha (T497) antibody (HA722278) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
☑ Cell treatment (CT)
Immunocytochemistry analysis of NIH/3T3 cells treated with or without λpp labeling Phospho-PKC alpha (T497) with Rabbit anti-Phospho-PKC alpha (T497) antibody (HA722278) at 1/500 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-PKC alpha (T497) antibody (HA722278) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
☑ Cell treatment (CT)
Immunocytochemistry analysis of C6 cells treated with or without λpp labeling Phospho-PKC alpha (T497) with Rabbit anti-Phospho-PKC alpha (T497) antibody (HA722278) at 1/500 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-PKC alpha (T497) antibody (HA722278) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Flow cytometric analysis of K-562 cells labeling Phospho-PKC alpha (T497).
Cells were fixed and permeabilized. Then stained with the primary antibody (HA722278, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
Flow cytometric analysis of NIH/3T3 cells labeling Phospho-PKC alpha (T497).
Cells were fixed and permeabilized. Then stained with the primary antibody (HA722278, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
Flow cytometric analysis of C6 cells labeling Phospho-PKC alpha (T497).
Cells were fixed and permeabilized. Then stained with the primary antibody (HA722278, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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Application: WB,IF-Cell,IF-Tissue,IHC-P,IP,Dot Blot
Reactivity: Human,Mouse,Rat
Conjugate: unconjugated
