Recombinant
RabMAb

Recombinant Anti-PKC beta 1 antibody [EPR18512] (ab195039)

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Overview

  • Product name

    Anti-PKC beta 1 antibody [EPR18512]
    See all PKC beta 1 primary antibodies

  • Description

    Rabbit monoclonal [EPR18512] to PKC beta 1

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human PKC beta 1 aa 600 to the C-terminus. The exact sequence is proprietary.
    Database link: P05771

  • Positive control

    • WB: Active human PKC beta 1 full length protein; Human fetal brain whole cell lysate; L-929, C2C12, U-87 MG, HeLa, C6 and NIH/3T3 cell lysates; Mouse brain and rat brain lysates. IHC-P: Human colon, Human colon cancer, mouse kidney and rat spleen tissues. ICC/IF: HeLa and K562 cells. Flow Cyt: K562 cells.

  • General notes

     

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.

  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR18512

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab195039 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 1/2000. Detects a band of approximately 77, 40 kDa (predicted molecular weight: 77 kDa).
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/100.
Flow Cyt 1/200.

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

    see all

  • Alternative names

    • KPCB_HUMAN antibody
    • PKC beta antibody
    • PKC-B antibody
    • PKC-beta antibody
    • PKCB antibody
    • Prkcb antibody
    • PRKCB I antibody
    • PRKCB1 antibody
    • PRKCB2 antibody
    • Protein kinase C beta antibody
    • Protein kinase C beta type antibody
    • protein kinase C, beta 1 polypeptide antibody
    • protein kinase C, beta-1 antibody
    see all

Images

  • Western blot - Anti-PKC beta 1 antibody [EPR18512] (ab195039)
    Western blot - Anti-PKC beta 1 antibody [EPR18512] (ab195039)
    All lanes : Anti-PKC beta 1 antibody [EPR18512] (ab195039) at 1/2000 dilution

    Lane 1 : Active human PKC alpha full length protein
    Lane 2 : Active human PKC beta 1 full length protein
    Lane 3 : Active human PKC beta 2 full length protein
    Lane 4 : Active human PKC gamma full length protein
    Lane 5 : Active human PKC delta full length protein
    Lane 6 : Active human PKC eta full length protein
    Lane 7 : Active human PKC epsilon full length protein
    Lane 8 : Active human PKC theta full length protein
    Lane 9 : Active human PKC mu full length protein

    Lysates/proteins at 0.02 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

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


    Exposure time: 1 second


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Active human PKC alpha full length protein (ab55672) contains aa1-672 with GST-tag; Active human PKC beta 1 full length protein (ab60840) contains aa1-671 with GST-tag; Active human PKC beta 2 full length protein (ab60841) contains aa1-673 with GST-tag; Active human PKC gamma full length protein (ab60842) contains aa1-677 with GST-tag; Active human PKC delta full length protein (ab60844) contains aa1-676 with GST-tag; Active human PKC eta full length protein (ab60849) contains aa1-683 with GST-tag; Active human PKC epsilon full length protein (ab60847) contains aa1-737 with GST-tag; Active human PKC theta full length protein (ab56641) contains aa1-706 with GST-tag; Active human PKC mu full length protein (ab60873) contains aa1-912 with GST-tag.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC beta 1 antibody [EPR18512] (ab195039)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC beta 1 antibody [EPR18512] (ab195039)

    Immunohistochemical analysis of paraffin-embedded human colon tissue labeling PKC beta 1 with ab195039 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus and weak cytoplasm staining on epithelial cells of Human colon is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

     

  • Immunocytochemistry/ Immunofluorescence - Anti-PKC beta 1 antibody [EPR18512] (ab195039)
    Immunocytochemistry/ Immunofluorescence - Anti-PKC beta 1 antibody [EPR18512] (ab195039)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling PKC beta 1 with ab195039 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and cytoplasm staining on K562 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab195039 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

     

  • Flow Cytometry - Anti-PKC beta 1 antibody [EPR18512] (ab195039)
    Flow Cytometry - Anti-PKC beta 1 antibody [EPR18512] (ab195039)

    Flow cytometric analysis of 4% paraformaldehyde-fixed K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling PKC beta 1 with ab195039 at 1/200 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.

  • Immunocytochemistry/ Immunofluorescence - Anti-PKC beta 1 antibody [EPR18512] (ab195039)
    Immunocytochemistry/ Immunofluorescence - Anti-PKC beta 1 antibody [EPR18512] (ab195039)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling PKC beta 1 with ab195039 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and cytoplasm staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab195039 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

     

  • Western blot - Anti-PKC beta 1 antibody [EPR18512] (ab195039)
    Western blot - Anti-PKC beta 1 antibody [EPR18512] (ab195039)
    All lanes : Anti-PKC beta 1 antibody [EPR18512] (ab195039) at 1/10000 dilution

    Lane 1 : Human fetal brain whole cell lysate
    Lane 2 : L-929 (Mouse connective tissue fibroblast cells) whole cell lysate
    Lane 3 : C2C12 (Mouse myoblast cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/50000 dilution

    Predicted band size: 77 kDa
    Observed band size: 40,77 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The peptide immunogen corresponds to the C terminal region of human PKC beta 1. The antibody recognizes the full length (~77KD) and the catalytic domain (~40KD) of PKC beta 1 (Geraldes P and King GL, 2010. Circ Res. 106, 1319-1331.

  • Western blot - Anti-PKC beta 1 antibody [EPR18512] (ab195039)
    Western blot - Anti-PKC beta 1 antibody [EPR18512] (ab195039)
    All lanes : Anti-PKC beta 1 antibody [EPR18512] (ab195039) at 1/10000 dilution

    Lane 1 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate
    Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 77 kDa
    Observed band size: 40,77 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The peptide immunogen corresponds to the C terminal region of human PKC beta 1. The antibody recognizes the full length (~77KD) and the catalytic domain (~40KD) of PKC beta 1 (Geraldes P and King GL, 2010. Circ Res. 106, 1319-1331.

  • Western blot - Anti-PKC beta 1 antibody [EPR18512] (ab195039)
    Western blot - Anti-PKC beta 1 antibody [EPR18512] (ab195039)
    All lanes : Anti-PKC beta 1 antibody [EPR18512] (ab195039) at 1/2000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Rat brain lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 77 kDa
    Observed band size: 40,77 kDa why is the actual band size different from the predicted?


    Exposure time: 15 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The peptide immunogen corresponds to the C terminal region of human PKC beta 1. The antibody recognizes the full length (~77KD) and the catalytic domain (~40KD) of PKC beta 1 (Geraldes P and King GL, 2010. Circ Res. 106, 1319-1331.

  • Western blot - Anti-PKC beta 1 antibody [EPR18512] (ab195039)
    Western blot - Anti-PKC beta 1 antibody [EPR18512] (ab195039)
    All lanes : Anti-PKC beta 1 antibody [EPR18512] (ab195039) at 1/2000 dilution

    Lane 1 : C6 (Rat glial tumor cells) whole cell lysate
    Lane 2 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 77 kDa
    Observed band size: 40,77 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The peptide immunogen corresponds to the C terminal region of human PKC beta 1. The antibody recognizes the full length (~77KD) and the catalytic domain (~40KD) of PKC beta 1 (Geraldes P and King GL, 2010. Circ Res. 106, 1319-1331.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC beta 1 antibody [EPR18512] (ab195039)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC beta 1 antibody [EPR18512] (ab195039)

    Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling PKC beta 1 with ab195039 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus and cytoplasm staining on cancer cells of colon cancer is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC beta 1 antibody [EPR18512] (ab195039)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC beta 1 antibody [EPR18512] (ab195039)

    Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling PKC beta 1 with ab195039 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus and cytoplasm staining on epithelial cells of mouse kidney is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC beta 1 antibody [EPR18512] (ab195039)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC beta 1 antibody [EPR18512] (ab195039)

    Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling PKC beta 1 with ab195039 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus and cytoplasm staining on lymphocytes of rat spleen is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

     

References

ab195039 has not yet been referenced specifically in any publications.

Publishing research using ab195039? Please let us know so that we can cite the reference in this datasheet.

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