Recombinant
RabMAb

Anti-PKC alpha + beta 2 + gamma antibody [EPR18104] (ab184746)

Overview

  • Product name
    Anti-PKC alpha + beta 2 + gamma antibody [EPR18104]
  • Description
    Rabbit monoclonal [EPR18104] to PKC alpha + beta 2 + gamma
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human PKC alpha + beta 2 + gamma aa 150-350. The exact sequence is proprietary.
    Database link: P05129

  • Positive control
    • WB: Active human PKC alpha, PKC gamma and PKC beta 2 full length proteins; Human fetal brain lysate; HeLa, Jurkat, K562, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Mouse and rat brain lysates. IHC-P: Human cerebral cortex, Human pancreas, Cervical cancer, Mouse kidney and Rat kidney tissues. ICC/IF: SH-SY5Y and HeLa cells. Flow Cyt: HeLa cells.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    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
    EPR18104
  • Isotype
    IgG

Applications

Our Abpromise guarantee covers the use of ab184746 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. Detects a band of approximately 78 kDa (predicted molecular weight: 77, 78 kDa).
IHC-P 1/400. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/100.
Flow Cyt 1/60.

Images

  • All lanes : Anti-PKC alpha + beta 2 + gamma antibody [EPR18104] (ab184746) at 1/1000 dilution

    Lane 1 : Active human PKC alpha full length protein
    Lane 2 : Active human PKC gamma full length protein

    Lysates/proteins at 0.02 µg per lane.

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

    Predicted band size : 77, 78 kDa


    Exposure time : 1 second

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Active human PKC alpha full length protein (Catalog#ab55672) contains aa1-672 with GST-tag; Active human PKC gamma full length protein (Catalog#ab60842) contains aa1-677 with GST-tag.

  • Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling PKC gamma with ab184746 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nucleus and weak cytoplasmic staining on Human pancreatic acinar cells is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SH-SY5Y (Human neuroblastoma cell line from bone marrow) cells labeling PKC gamma with ab184746 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing cytoplasmic and nuclear staining on SH-SY5Y cell line. 

    The nuclear counterstain 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: ab184746 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 cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling PKC gamma with ab184746 at 1/60 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.

  • All lanes : Anti-PKC alpha + beta 2 + gamma antibody [EPR18104] (ab184746) at 1/1000 dilution

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

    Lysates/proteins at 0.02 µg per lane.

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

    Predicted band size : 77, 78 kDa


    Exposure time : 3 minutes

    Blocking/Dilution buffer: 5% NFDM/TBST.

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

  • Anti-PKC alpha + beta 2 + gamma antibody [EPR18104] (ab184746) at 1/1000 dilution + Active human PKC beta 1 full length protein at 0.02 µg

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

    Predicted band size : 77, 78 kDa


    Exposure time : 3 minutes

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Active human PKC beta 1 full length protein (Catalog #ab60840) contains aa1-671 with GST-tag.

  • Anti-PKC alpha + beta 2 + gamma antibody [EPR18104] (ab184746) at 1/1000 dilution + Human fetal brain lysate at 10 µg

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size : 77, 78 kDa
    Observed band size : 78 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 second

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-PKC alpha + beta 2 + gamma antibody [EPR18104] (ab184746) at 1/1000 dilution

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
    Lane 3 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

    Predicted band size : 77, 78 kDa
    Observed band size : 78 kDa (why is the actual band size different from the predicted?)


    Exposure time : 2 seconds

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-PKC alpha + beta 2 + gamma antibody [EPR18104] (ab184746) at 1/1000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Rat brain lysate
    Lane 3 : C6 (Rat glial tumor cell line) whole cell lysate
    Lane 4 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 5 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 6 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

    Predicted band size : 77, 78 kDa
    Observed band size : 78 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 second

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling PKC gamma with ab184746 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing cytoplasmic and nuclear staining on HeLa cell line.

    The nuclear counterstain 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: ab184746 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.

  • Immunohistochemical analysis of paraffin-embedded cervical cancer tissue labeling PKC gamma with ab184746 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nucleus and weak cytoplasmic staining on cancer cells of cervix cancer is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling PKC gamma with ab184746 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasmic 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 ab97051 at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling PKC gamma with ab184746 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nucleus and cytoplasmic staining on epithelial cells of rat kidney is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded human cerebral cortex tissue labeling PKC gamma with ab184746 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Weak cytoplasmic staining on neuron of Human cerebral cortex 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.

References

ab184746 has not yet been referenced specifically in any publications.

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