Q: We are looking into protein kinase C-alpha antibodies for immunohistochemistry on formalin fixed, paraffin embedded tissue. I see that you have many antibodies, including several monoclonals. I wanted to confirm with you that the following antibodies are specific for C-alpha. NB600-201=MC5 (an older paper suggested that this might react with alpha, beta, gamm), NB110-57356=Y124, NB110-57353=Y143. Also, if you have an comparator data or feedback regarding PKC-alpha antibodies for IHC on FFPE, that would be very helpful. We would like to pick 1 antibody that is easy to titer and robust, so that we can move forward with our studies.

A: We have never heard any feedback that these cross-react with other PKC proteins. If you have seen data that MC5 may cross-react, then I would consider avoiding this antibody if you are concerned. The immunogens for NB110-57356 and NB110-57353 come from the N- and C-terminals, which is where the PKC proteins vary the most, so these are not expected to cross-react and we have had no feedback or testing data that suggests that they do. I would recommend NB110-57356, since it has been published with. You may want to review the published data to determine if it seems this antibody will suit your needs.

PKC alpha [p Ser657, p Tyr658] Antibody

Novus Biologicals | Catalog # NBP3-23303

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Product Documents

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Product Specifications
Scientific Data
Applications
Preparation & Storage
Background
Product Documents
Specific Notices
Research Areas

Key Product Details

Validated by

Biological Validation

Species Reactivity

Human, Rat, Rabbit

Applications

Western Blot, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

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Product Specifications

Immunogen

Phospho-PKC alpha (Ser-657/Tyr-658) synthetic peptide corresponds to amino acid residues around serine 657 and tyrosine 658 of human PKC alpha. This sequence is similar to the conserved sites in rat and mouse, and has high homology to dual sites in human, rat, and mouse PKC-beta-I (Ser-661/Tyr-662) and PKC gamma (Thr-674/Tyr-675). [UniProt# P17252]

Modification

p Ser657, p Tyr658

Specificity

This antibody detects an 82kDa protein corresponding to the molecular mass of phosphorylated PKC alpha on SDS-PAGE immunoblots of neonatal rat brain lysate. Similar results were observed in human SKN-SH, endothelial, and HeLa cells, as well as rabbit spleen fibroblasts and rat pituitary cells. In immunoprecipitation experiments with various PKC isoforms, this antibody detected PKC alpha and PKC beta but not other PKC isoforms in rat brain lysate.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

82 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Description

Stable for at least 1 year at -20°C.

Scientific Data Images for PKC alpha [p Ser657, p Tyr658] Antibody

Immunocytochemistry/Immunofluorescence: PKC alpha [p Ser657, p Tyr658] Antibody [NBP3-23303] -

Immunocytochemistry/Immunofluorescence: PKC alpha [p Ser657, p Tyr658] Antibody [NBP3-23303] - Immunocytochemical labeling of PKC phosphorylation in aldehyde-fixed and NP-40-permeabilized NGF-differentiated PC12 cells. The cells were labeled with rabbit polyclonal anti-PKC alpha (Ser-657/Tyr-658) (NBP3-23303) antibody in the absence (Left) or presence (Right) of blocking peptide. The antibody was detected using appropriate secondary antibody conjugated to DyLight(R) 594.

Western Blot: PKC alpha [p Ser657, p Tyr658] Antibody [NBP3-23303] -

Western Blot: PKC alpha [p Ser657, p Tyr658] Antibody [NBP3-23303] - Western blot analysis of immunoprecipitates from neonatal rat brain lysate using anti-PKC alpha antibody. Control and alkaline phosphatase treated precipitates were probed with anti-PKC alpha (Central region) or anti-phospho-PKC alpha (Ser-657/Tyr-658). The latter shows no detection of PKC alpha after phosphatase treatment.

Applications for PKC alpha [p Ser657, p Tyr658] Antibody

Application

Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:100

Immunoprecipitation

1:100

Western Blot

1:1000

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Formulation

PBS, 1 mg/ml BSA and 50% glycerol

Preservative

0.05% Sodium Azide

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C.

Background: PKC alpha

Protein Kinase C (PKC) is a large superfamily of serine/threonine kinases that mediate essential cellular signals required for activation, proliferation, differentiation and survival. There are at least ten PKC isotypes that are closely related in structure but that have distinct patterns of tissue distribution and function. The PKC isotypes can be subdivided into three classes based on primary structure and biochemical properties. These are: classical or conventional PKC isotypes (cPKC), novel PKC isotypes (nPKC) and atypical PKC isotypes (aPKC). All PKC isotypes share a characteristic equence motif C1 in addition to a serine/threonine-protein kinase domain. The cPKC isotypes include PKC

Long Name

Protein Kinase C alpha

Alternate Names

PKCA, PRKCA

Gene Symbol

PRKCA

Additional PKC alpha Products

Product Documents for PKC alpha [p Ser657, p Tyr658] Antibody

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Certificate of Analysis

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Product Specific Notices for PKC alpha [p Ser657, p Tyr658] Antibody

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Related Research Areas

Diabetes

Intracellular Kinases

PDGF Family

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Protocols

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FAQs for PKC alpha [p Ser657, p Tyr658] Antibody

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Q: We are looking into protein kinase C-alpha antibodies for immunohistochemistry on formalin fixed, paraffin embedded tissue. I see that you have many antibodies, including several monoclonals. I wanted to confirm with you that the following antibodies are specific for C-alpha. NB600-201=MC5 (an older paper suggested that this might react with alpha, beta, gamm), NB110-57356=Y124, NB110-57353=Y143. Also, if you have an comparator data or feedback regarding PKC-alpha antibodies for IHC on FFPE, that would be very helpful. We would like to pick 1 antibody that is easy to titer and robust, so that we can move forward with our studies.

A:
We have never heard any feedback that these cross-react with other PKC proteins. If you have seen data that MC5 may cross-react, then I would consider avoiding this antibody if you are concerned. The immunogens for NB110-57356 and NB110-57353 come from the N- and C-terminals, which is where the PKC proteins vary the most, so these are not expected to cross-react and we have had no feedback or testing data that suggests that they do. I would recommend NB110-57356, since it has been published with. You may want to review the published data to determine if it seems this antibody will suit your needs.