Recombinant
RabMAb

Anti-PKC beta 2 antibody [Y125] (ab32026)

Overview

  • Product name
    Anti-PKC beta 2 antibody [Y125]
    See all PKC beta 2 primary antibodies
  • Description
    Rabbit monoclonal [Y125] to PKC beta 2
  • Specificity
    ab32026 recognises PKC beta 2, it does not cross react with human PKC beta 1. Immunogen's homology with PKC alpha is 77.8%. Cross-reactivity with PKC alpha has not been tested.
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details
  • Species reactivity
    Reacts with: Rat, Human
    Predicted to work with: Cow, PigDoes not react with: Mouse
  • Immunogen

    Synthetic peptide corresponding to residues near C terminus of human PKC beta 2

  • Epitope
    ab32026 reacts with an epitope located in the region near the C terminus of PKC beta 2.
  • Positive control
    • K562 cell lysate, HeLa cell lysate, rat spinal cord lysate and human breast carcinoma
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    A trial size is available to purchase for this antibody.

    Alternative versions available:

    Anti-PKC beta 2 antibody (Alexa Fluor® 488) [Y125] (ab195251)

    Anti-PKC beta 2 antibody (Alexa Fluor® 647) [Y125] (ab195252)

    Anti-PKC beta 2 antibody (HRP) [Y125] (ab194906)

    Anti-PKC beta 2 antibody (Phycoerythrin) [Y125] (ab210976)

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer
    PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    Y125
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab32026 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000 - 1/10000. Detects a band of approximately 77 kDa (predicted molecular weight: 77 kDa).
IHC-P Use at an assay dependent concentration.
ICC/IF 1/250 - 1/500.
Flow Cyt 1/50.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP 1/60.

Target

  • Function
    Calcium-activated and phospholipid-dependent serine/threonine-protein kinase involved in various processes such as regulation of the B-cell receptor (BCR) signalosome, apoptosis and transcription regulation. Plays a key role in B-cell activation and function by regulating BCR-induced NF-kappa-B activation and B-cell suvival. Required for recruitment and activation of the IKK kinase to lipid rafts and mediates phosphorylation of CARD11/CARMA1 at 'Ser-559', 'Ser-644' and 'Ser-652', leading to activate the NF-kappa-B signaling. Involved in apoptosis following oxidative damage: in case of oxidative conditions, specifically phosphorylates 'Ser-36' of isoform p66Shc of SHC1, leading to mitochondrial accumulation of p66Shc, where p66Shc acts as a reactive oxygen species producer. Acts as a coactivator of androgen receptor (ANDR)-dependent transcription, by being recruited to ANDR target genes and specifically mediating phosphorylation of 'Thr-6' of histone H3 (H3T6ph), a specific tag for epigenetic transcriptional activation that prevents demethylation of histone H3 'Lys-4' (H3K4me) by LSD1/KDM1A. Also involved in triglyceride homeostasis. Serves as the receptor for phorbol esters, a class of tumor promoters.
  • Sequence similarities
    Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. PKC subfamily.
    Contains 1 AGC-kinase C-terminal domain.
    Contains 1 C2 domain.
    Contains 2 phorbol-ester/DAG-type zinc fingers.
    Contains 1 protein kinase domain.
  • Post-translational
    modifications
    Phosphorylation on Thr-500 within the activation loop renders it competent to autophosphorylate. Subsequent autophosphorylation of Thr-642 maintains catalytic competence, and autophosphorylation on Ser-661 appears to release the kinase into the cytosol. Autophosphorylation on other sites i.e. in the N-terminal and hinge regions have no effect on enzyme activity.
  • Cellular localization
    Cytoplasm. Nucleus. Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • KPCB_HUMAN antibody
    • PKC Beta antibody
    • PKC-B antibody
    • PKC-beta antibody
    • PKCB antibody
    • Prkcb antibody
    • PRKCB II antibody
    • PRKCB2 antibody
    • Protein kinase C beta antibody
    • Protein kinase C beta type antibody
    see all

Anti-PKC beta 2 antibody [Y125] images

  • Anti-PKC beta 2 antibody [Y125] (ab32026) at 1/500 dilution + Recombinant human PKC beta 2 protein (ab60841) at 0.01 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 77 kDa
    Observed band size : 103 kDa (why is the actual band size different from the predicted?)


    Exposure time : 20 seconds

    Blocking Buffer: 5% NFDM/TBST

    Diluting Buffer: 5% NFDM/TBST

  • ab32026, at a 1/100 dilution, staining human PKC beta 2 in breast carcinoma by Immunohistochemistry, Paraffin embedded tissue

  • ab32026, at a 1/250 dilution, staining human PKC beta 2 in HeLa cells by Immunoflourescence
  • Overlay histogram showing HeLa cells stained with ab32026 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32026, 1/50 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • All lanes : Anti-PKC beta 2 antibody [Y125] (ab32026) at 1/500 dilution

    Lane 1 : K562 whole cell lysates at 20 µg
    Lane 2 : Active human PKC beta 2 full length protein at 0.02 µg
    Lane 3 : Human fetal brain at 20 µg
    Lane 4 : NIH/3T3 whole cell lysates at 20 µg
    Lane 5 : Raw264.7 whole cell lysates at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 77 kDa
    Observed band size : 77 kDa


    Exposure time : 15 seconds

    Blocking buffer: 5% NFDM/TBST

    Diluting buffer: 5% NFDM/TBST

    The PKC beta 2 recombinant protein is GST-tagged, so the molecular weight is higher.

  • Anti-PKC beta 2 antibody [Y125] (ab32026) at 1/500 dilution + Rat spinal cord at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 77 kDa
    Observed band size : 77 kDa


    Exposure time : 3 minutes

    Blocking Buffer: 5% NFDM/TBST

    Diluting Buffer: 5% NFDM/TBST

  • Anti-PKC beta 2 antibody [Y125] (ab32026) at 1/500 dilution + K562 cell lysate

    Predicted band size : 77 kDa
    Observed band size : 77 kDa
  • All lanes : Anti-PKC beta 2 antibody [Y125] (ab32026) at 1/1000 dilution

    Lane 1 : MCF-7 whole cell lysates
    Lane 2 : HeLa whole cell lysates
    Lane 3 : K562 whole cell lysates
    Lane 4 : Recombinant human PKC alpha protein (ab55672)

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 77 kDa
    Observed band size : 77 kDa


    Exposure time : 30 seconds

    Blocking Buffer: 5% NFDM/TBST

    Diluting Buffer: 5% NFDM/TBST

References for Anti-PKC beta 2 antibody [Y125] (ab32026)

This product has been referenced in:
  • Ravindran R  et al. The amino acid sensor GCN2 controls gut inflammation by inhibiting inflammasome activation. Nature 531:523-7 (2016). IHC-P ; Mouse . Read more (PubMed: 26982722) »
  • Braggin JE  et al. Essential role of protein kinase R antagonism by TRS1 in human cytomegalovirus replication. Virology 489:75-85 (2016). Read more (PubMed: 26716879) »

See all 7 Publications for this product

Product Wall

Application
Western blot
Sample
Cow Cell lysate - other (Cerebral microvascular endothelial cells)
Gel Running Conditions
Reduced Denaturing (8% Bis-Tris)
Loading amount
10 µg
Specification
Cerebral microvascular endothelial cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 7.5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Apr 14 2015

Thank you for contacting us.

This antibody has been tested in IP so it will more likely to work in Co-IP also. However we cannot confirm it because we do not have tested data. The experimental conditions have to be determined by the end user...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (MCF7)
Loading amount
100000 cells
Specification
MCF7
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Abcam user community

Verified customer

Submitted May 12 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (GLC16 Cells)
Loading amount
40000 cells
Specification
GLC16 Cells
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Feb 10 2010

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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