Anti-PKC beta 1 (phospho T642) antibody (ab5782)
Key features and details
- Rabbit polyclonal to PKC beta 1 (phospho T642)
- Suitable for: WB
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-PKC beta 1 (phospho T642) antibody
See all PKC beta 1 primary antibodies -
Description
Rabbit polyclonal to PKC beta 1 (phospho T642) -
Host species
Rabbit -
Specificity
This antibody does not react with PKC beta 2 [pT641], alpha [pT638], nu [pT655], epsilon [pT710], iota [pT555], eta [pT655], or zeta [pT560] 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 that contains threonine 642.
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Positive control
- K562 cells treated with PMA, a phorbol ester.
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 (glycerin, glycerine), 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 1. The final product is generated by affinity chromatography using a PKC beta 1-derived peptide that is phosphorylated at threonine 642. -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Images
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Western blot - Anti-PKC beta 1 (phospho T642) antibody (ab5782)
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 (ë) phosphatase (5), blocked with a 5% BSA-TBST buffer for one hour at room temperature, and incubated with ab5782 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 I [pT642] 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 (ë) phosphatase (5), blocked with a 5% BSA-TBST buffer for one hour at room temperature, and incubated with ab5782 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 I [pT642] blocks the antibody signal. The data also show that phosphatase stripping eliminates the signal, verifying that the antibody is phospho-specific.
