Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-PKC delta antibody [EPR17075] - BSA and Azide free (ab222229)

Overview

  • Product name

    Anti-PKC delta antibody [EPR17075] - BSA and Azide free
    See all PKC delta primary antibodies
  • Description

    Rabbit monoclonal [EPR17075] to PKC delta - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment aa 500 to the C-terminus. The exact sequence is proprietary.
    Database link: P28867

  • Positive control

    • WB: A431, C6, NIH/3T3 and HeLa whole cell lysates, human fetal brain and fetal heart, mouse and rat thymus and brain, and rat spleen tissue lysates. IHC-P: Human spleen, human transitional cell carcinoma of bladder, mouse liver and rat testis tissues. ICC/IF: Wild-type HAP1 (PMA-treated and untreated) and HeLa cells. Flow Cyt: HeLa cells.
  • General notes

    Ab222229 is the carrier-free version of ab182126. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab222229 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab222229 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 Use at an assay dependent concentration. Detects a band of approximately 40, 78 kDa (predicted molecular weight: 78 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration.

Treatment with 10nM PMA for 10 min induces translocation of PKCδ to the membrane. 

Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • Function

    This is calcium-independent, phospholipid-dependent, serine- and threonine-specific enzyme. PKC is activated by diacylglycerol which in turn phosphorylates a range of cellular proteins. PKC also serves as the receptor for phorbol esters, a class of tumor promoters. May play a role in antigen-dependent control of B-cell function. Phosphorylates MUC1 in the C-terminal and regulates the interaction between MUC1 and beta-catenin.
  • 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.
  • Domain

    The C1 domain, containing the phorbol ester/DAG-type region 1 (C1A) and 2 (C1B), is the diacylglycerol sensor.
    The C2 domain is a non-calcium binding domain. It binds proteins containing phosphotyrosine in a sequence-specific manner.
  • Post-translational
    modifications

    Phosphorylated on Thr-507, within the activation loop. Autophosphorylated and/or phosphorylated. Although the Thr-507 phosphorylation occurs it is not a prerequisite for enzymatic activity.
  • Cellular localization

    Cytoplasm. Membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • CVID9 antibody
    • D14Ertd420e antibody
    • Kinase PKC delta antibody
    • KPCD antibody
    • KPCD_HUMAN antibody
    • MAY 1 antibody
    • MAY1 antibody
    • MGC49908 antibody
    • nPKC delta antibody
    • nPKC-delta antibody
    • PCKd antibody
    • PKC d antibody
    • PKC delta antibody
    • PKCD antibody
    • PKCdelta antibody
    • PRKC D antibody
    • PRKC delta antibody
    • Prkcd antibody
    • Protein Kinase C delta antibody
    • Protein kinase C delta type antibody
    • Protein kinase C delta VIII antibody
    • Protein Kinase Cdelta antibody
    • Tyrosine protein kinase PRKCD antibody
    see all

Images

  • Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling PKC delta with purified ab182126 at 1/250 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182126).

  • ab182126 staining PKCδ in untreated wild-type HAP1 cells (top panel) and PKCδ untreated knockout HAP1 cells (bottom panel). Untreated cells show PKCδ being expressed in the cytoplasm. The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab182126 at 1/200 dilution and ab7291 at 1ug/ml concentration overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to Mouse IgG (Alexa Fluor® 594) (ab150117) at 2ug/ml (shown in pseudo-color red). Nuclear DNA was labelled in blue with DAPI.

    This product also gave a positive signal under the same testing conditions in HAP1 cells fixed with 100% methanol (5 min).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182126).

  • ab182126 staining PKCδ in 10nM PMA-treated wild-type HAP1 cells (top panel) and PKCδ in 10nM PMA-treated knockout HAP1 cells (bottom panel). The cells were treated with 10nM PMA for 10 minutes to induce translocation of PKCδ to the cell membrane. The cells were then fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab182126 at 1/200 dilution and ab7291 at 1ug/ml concentration overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to Mouse IgG (Alexa Fluor® 594) (ab150117) at 2ug/ml (shown in pseudo-color red). Nuclear DNA was labelled in blue with DAPI.

    This product also gave a positive signal under the same testing conditions in HAP1 cells fixed with 100% methanol (5 min).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182126).

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling PKC delta with ab182126 at 1/100 dilution. The cells were permeabilised with 0.1% Triton X-100. Goat anti-rabbit IAlexa Fluor® 488 (IgG) (ab150077) at 1/400 dilution was used as the secondary antibody (green). The confocal image shows both cytoplasmic and nuclear staining on HeLa cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 and ab150120 (goat anti-mouse AlexaFluor® 594 secondary antibody) at 1/500 dilution (red).
    The negative controls are as follows;
    1. ab182126 at 1/100 dilution followed by ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/500 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor® 488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182126).

  • Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling PKC delta with ab182126 at 1/2000 dilution, followed by Anti-Rabbit HRP (ab97051) at 1/500 dilution. Cytoplasmic and nucleus staining on lymphocytes of Human spleen is detected. The negative control utilised PBS instead of primary antibody. Counter stained with Hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182126).

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

  • Immunohistochemical analysis of paraffin-embedded Human transitional cell carcinoma of bladder tissue labeling PKC delta with ab182126 at 1/2000 dilution, followed by Anti-Rabbit HRP (ab97051) at 1/500 dilution. Cytoplasmic and weak nuclear staining on cancer cells of bladder transitional cell carcinoma is detected. The negative control utilised PBS instead of primary antibody. Counter stained with Hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182126).

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

  • Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling PKC delta with ab182126 at 1/2000 dilution, followed by Anti-Rabbit HRP (ab97051) at 1/500 dilution. Cytoplasmic and nuclear staining on kupffer cells of Mouse liver is detected. The negative control utilised PBS instead of primary antibody. Counter stained with Hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182126).

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

  • Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling PKC delta with ab182126 at 1/2000 dilution, followed by Anti-Rabbit HRP (ab97051) at 1/500 dilution. Cytoplasmic and weak nuclear staining on cells in the seminiferous tubule of rat testis is detected. The negative control utilised PBS instead of primary antibody. Counter stained with Hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182126).

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

References

ab222229 has not yet been referenced specifically in any publications.

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