PKC Alpha Monoclonal antibody

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• KD/KO Validated

PKC Alpha Monoclonal Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat

Applications

WB, IHC, IF/ICC, FC (Intra), ELISA

Conjugate

Unconjugated

Publications

1

CloneNo.

1D11F4

Cat no : 66421-1-Ig

Synonyms

PRKCA, PKC-A, PKCA, PKC A, EC:2.7.11.13

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Validation Data Gallery

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  WB analysis using 66421-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66421-1-Ig (PKC Alpha antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis of HEK-293 using 66421-1-Ig

  WB result of PKC Alpha antibody (66421-1-Ig; 1:8000; incubated at room temperature for 1.5 hours) with sh-Control and sh-PKC Alpha transfected HEK-293 cells.

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  WB analysis of HeLa using 66421-1-Ig

  HeLa cells were subjected to SDS PAGE followed by western blot with 66421-1-Ig (PKC alpha antibody at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis of HEK-293 using 66421-1-Ig

  HEK-293 cells were subjected to SDS PAGE followed by western blot with 66421-1-Ig (PKC alpha antibody at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis of C6 using 66421-1-Ig

  C6 cells were subjected to SDS PAGE followed by western blot with 66421-1-Ig (PKC alpha antibody at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis of RAW 264.7 using 66421-1-Ig

  RAW 264.7 cells were subjected to SDS PAGE followed by western blot with 66421-1-Ig (PKC alpha antibody at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  IHC staining of human lung cancer using 66421-1-Ig

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 66421-1-Ig (PKC alpha antibody at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human lung cancer using 66421-1-Ig

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 66421-1-Ig (PKC alpha antibody at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HeLa using 66421-1-Ig

  Immunofluorescent analysis of (-20°C Methanol) fixed HeLa cells using PKC Alpha antibody (66421-1-Ig, Clone: 1D11F4 ) at dilution of 1:800 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L).

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  IF Staining of HeLa using 66421-1-Ig

  Immunofluorescent analysis of (-20°C Ethanol) fixed HeLa cells using PKC Alpha antibody (66421-1-Ig, Clone: 1D11F4 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L).

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  IF Staining of HeLa using 66421-1-Ig

  Immunofluorescent analysis of (-20℃ Ethanol ) fixed HeLa cells using 66421-1-Ig(PKC alpha antibody) at dilution of 1:100 and Alexa Fluor 488-conjugated Goat Anti-Mouse IgG(H+L).

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  FC experiment of HeLa using 66421-1-Ig

  1X10^6 HeLa cells were intracellularly stained with 0.4 ug Anti-Human PKC Alpha (66421-1-Ig, Clone:1D11F4) and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Mouse IgG1 Isotype Control (MOPC-21) (65124-1-Ig, Clone: MOPC-21) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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Tested Applications

• Positive WB detected in: HeLa cells, HEK-293 cells, C6 cells, RAW 264.7 cells, HepG2 cells, Jurkat cells, K-562 cells, HSC-T6 cells, NIH/3T3 cells
• Positive IHC detected in: human lung cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HeLa cells
• Positive FC (Intra) detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:400-1:1600
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 1 publications below

Product Information

66421-1-Ig targets PKC Alpha in WB, IHC, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: mouse
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: PKC Alpha fusion protein Ag17275
• Full Name: protein kinase C, alpha
• Observed Molecular Weight: 77 kDa
• GenBank Accession Number: BC109274
• Gene Symbol: PKC Alpha
• Gene ID (NCBI): 5578
• RRID: AB_2881793
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

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. PRKCA 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 Ca(2+) handling in myocytes.

Publications

Species	Application	Title
mouse	WB	Ecotoxicol Environ Saf; Arsenic interferes with spermatogenesis involving Rictor/mTORC2-mediated blood-testis barrier disruption in mice; Authors - Xiujuan Li