PKC alpha Overexpression Lysate

Novus Biologicals | Catalog # NBP2-04234

Novus Biologicals
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Key Product Details

Species

Human

Applications

Western Blot

Product Summary for PKC alpha Overexpression Lysate

PKC alpha Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC211893

Accession#: NM_002737

Protein Tag: C-MYC/DDK

You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.

Product Specifications

Application Notes

This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.

Theoretical MW

76.6 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.

Type

Overexpression

Scientific Data Images for PKC alpha Overexpression Lysate

Western Blot: PKC alpha Overexpression Lysate [NBP2-04234]

Western Blot: PKC alpha Overexpression Lysate [NBP2-04234]

Western Blot: PKC alpha Overexpression Lysate (Adult Normal) [NBP2-04234] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for PKC alpha.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

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 Overexpression Lysate

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Product Specific Notices for PKC alpha Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Related Research Areas

Diabetes
Intracellular Kinases
PDGF Family

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FAQs for PKC alpha Overexpression Lysate

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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.

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