Anti-PKC beta 1 + PKC beta 2 (phospho T500) antibody (ab5817)
Key features and details
- Rabbit polyclonal to PKC beta 1 + PKC beta 2 (phospho T500)
- Suitable for: WB
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-PKC beta 1 + PKC beta 2 (phospho T500) antibody
See all PKC beta 1 + PKC beta 2 primary antibodies -
Description
Rabbit polyclonal to PKC beta 1 + PKC beta 2 (phospho T500) -
Host species
Rabbit -
Specificity
This antibody cross-reacts with PKC alpha [pT497] (88% homologous) and partially cross-reacts with PKC gamma [pT514] (63% homologous) and epsilon [pT566] (75% homologous), as determined by peptide competition experiments. -
Tested Applications & Species
See all applications and species dataApplication Species WB Human
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Immunogen
Synthetic phosphopeptide derived from a region of human PKC beta 1 & 2 that contains threonine 500.
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General notes
Protein Kinase C beta (PKC beta) is an 80 kDa member of the conventional group (cPKCs: sensitive to diacylglycerol, phosphotidylserine 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, transcriptional regulation and tumor promotion. PKC beta has been implicated in diabetes and carcinogenesis. PKC beta isoforms 1 & 2 are phosphorylated on three sites, threonine 500 in the activation loop, beta 1 threonine 642 (beta 2 641) in the turn loop and beta 1 serine 661 (beta 2 660) in the hydrophobic loop. Phosphorylation of PKC beta 1 & 2 on threonine 500 by PDK1 is a prerequisite for its autophosphoylation and catalytic competence.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
Storage buffer
pH: 7.30
Preservative: 0.05% Sodium azide
Constituents: PBS, 50% Glycerol, 0.1% BSA -
Concentration information loading... -
Purity
Immunogen affinity purified -
Purification notes
The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated PKC beta. The final product is generated by affinity chromatography using a PKC beta-derived peptide that is phosphorylated at threonine 500. -
Primary antibody notes
Protein Kinase C beta (PKC beta) is an 80 kDa member of the conventional group (cPKCs: sensitive to diacylglycerol, phosphotidylserine 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, transcriptional regulation and tumor promotion. PKC beta has been implicated in diabetes and carcinogenesis. PKC beta isoforms 1 & 2 are phosphorylated on three sites, threonine 500 in the activation loop, beta 1 threonine 642 (beta 2 641) in the turn loop and beta 1 serine 661 (beta 2 660) in the hydrophobic loop. Phosphorylation of PKC beta 1 & 2 on threonine 500 by PDK1 is a prerequisite for its autophosphoylation and catalytic competence. -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Images
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Western blot - Anti-PKC beta 1 + PKC beta 2 (phospho T500) antibody (ab5817)Peptide Competition and Phosphatase Treatment: Lysates prepared from K562 cells stimulated with PMA were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were either left untreated (1-4) or treated with lambda phosphatase (5), blocked with a 5% BSA-TBST buffer for one hour at room temperature, and incubated with ab5817 antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 5), the non-phosphopeptide corresponding to the immunogen (2), a generic phosphothreonine-containing peptide (3), or, the phosphopeptide immunogen (4). After washing, membranes were incubated with goat F(ab’ 2 anti-rabbit IgG HRP conjugate and bands were detected using the Pierce SuperSignalTM method. The data show that only the peptide corresponding to PKC beta 1 & 2 [pT500] blocks the antibody signal. The data also show that phosphatase stripping eliminates the signal, verifying that the antibody is phospho-specific.
Peptide Competition and Phosphatase Treatment: Lysates prepared from K562 cells stimulated with PMA were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were either left untreated (1-4) or treated with lambda phosphatase (5), blocked with a 5% BSA-TBST buffer for one hour at room temperature, and incubated with ab5817 antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 5), the non-phosphopeptide corresponding to the immunogen (2), a generic phosphothreonine-containing peptide (3), or, the phosphopeptide immunogen (4). After washing, membranes were incubated with goat F(ab’ 2 anti-rabbit IgG HRP conjugate and bands were detected using the Pierce SuperSignalTM method. The data show that only the peptide corresponding to PKC beta 1 & 2 [pT500] blocks the antibody signal. The data also show that phosphatase stripping eliminates the signal, verifying that the antibody is phospho-specific.
