Overview

  • Product nameAnti-PKC alpha antibody [Y124]
    See all PKC alpha primary antibodies
  • Description
    Rabbit monoclonal [Y124] to PKC alpha
  • Tested applicationsSuitable for: WB, IHC-P, Flow Cyt, IP, ICC/IF, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Pig, Goldfish
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human PKC alpha aa 650 to the C-terminus (C terminal). The exact sequence is proprietary.

  • Positive control
    • WB: 293 cell lysate. IHC-P: Human lung carcinoma. IF: PMA-Treated and untreated HAP1-PRKCA cells (wild-type and knockout), HeLa cells.
  • General notes

    A trial size is available to purchase for this antibody.

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab32376 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 75 kDa (predicted molecular weight: 77 kDa).
IHC-P 1/250 - 1/500.
Flow Cyt 1/100.

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

IP 1/100.
ICC/IF 1/200.

Treatment with 200nM PMA for 30 minutes induces translocation of PKC alpha to the membrane.

IHC-Fr 1/100.

Target

  • FunctionThis is a calcium-activated, phospholipid-dependent, serine- and threonine-specific enzyme. May play a role in cell motility by phosphorylating CSPG4.
    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.
  • Sequence similaritiesBelongs 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.
  • Cellular localizationCytoplasm. Cell membrane. Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • AAG6 antibody
    • Aging associated gene 6 antibody
    • aPKC antibody
    • KPCA_HUMAN antibody
    • PKC alpha antibody
    • PKC-A antibody
    • PKC-alpha antibody
    • PKCA antibody
    • PRKACA antibody
    • PRKCA antibody
    • Protein Kinase C alpha antibody
    • Protein kinase C alpha type antibody
    see all

Anti-PKC alpha antibody [Y124] images



  • Predicted band size : 77 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: PKC alpha knockout HAP1 cell lysate (20 µg)
    Lane 3: K562 cell lysate (20 µg)
    Lane 4: HEK293 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab32376 observed at 77 kDa. Red - loading control, ab8245, observed at 37 kDa.


    ab32376 was shown to specifically react with PKC alpha when PKC alpha knockout samples were used. Wild-type and PKC alpha knockout samples were subjected to SDS-PAGE. ab32376 and ab8245 (loading control to GAPDH) were diluted 1/5000 and 1/1000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) ab216776 secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • ab32376 staining PKCα in 200nM PMA-treated wild-type HAP1 cells (top panel) and PKCα in 200nM PMA-treated knockout HAP1 cells (bottom panel). The cells were treated with 200nM PMA for 30 minutes to induce translocation of PKCα to the cell membrane. The cells were then fixed with 100% methanol (5min), 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 ab32376 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 4% formaldehyde (10 min).


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

  • ab32376 staining PKCα in wild-type HAP1 cells (top panel) and PKCα in knockout HAP1 cells (bottom panel). In untreated conditions, PKCα is expressed in the cytoplasm of the cells. The cells were then fixed with 100% methanol (5min), 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 ab32376 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 4% formaldehyde (10 min).


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

  • Ab32376, at a 1/250 dilution, staining PKC alpha in paraffin embedded human lung carcinoma tissue sections by Immunohistochemistry.
  • Ab32376, at a 1/250 dilution, staining PKC alpha in HeLa cells by Immunofluorescence.
  • Overlay histogram showing HeLa cells stained with ab32376 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab32376, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • Anti-PKC alpha antibody [Y124] (ab32376) at 1/10000 dilution + 293 cell lysate

    Predicted band size : 77 kDa
    Observed band size : 75 kDa (why is the actual band size different from the predicted?)
  • Immunohistochemical analysis of mouse small intestine tissue, staining PKC alpha with ab32376.

References for Anti-PKC alpha antibody [Y124] (ab32376)

This product has been referenced in:
  • Wang XH  et al. Cannabinoid CB1 receptor signaling dichotomously modulates inhibitory and excitatory synaptic transmission in rat inner retina. Brain Struct Funct 221:301-16 (2016). Read more (PubMed: 25273281) »
  • Lum MA  et al. PKCa is Resistant to Long-Term Desensitization/Downregulation by Prolonged Diacylglycerol Stimulation. J Biol Chem N/A:N/A (2016). WB, IHC, ICC/IF ; Rat, Human . Read more (PubMed: 26769967) »

See all 24 Publications for this product

Product Wall

Application Western blot
Sample Rat Tissue lysate - whole (heart)
Gel Running Conditions Reduced Denaturing (9%)
Loading amount 60 µg
Treatment TNF-a for 30in
Specification heart
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Mr. Baptiste Jude

Verified customer

Submitted Jul 07 2016

Sorry for the delay in getting back to you.

The peptide used to raise the anti-PKC alpha antibody [Y124] (ab32376) has now been added to the catalogue for you. This is under the catalogue number ab133953.

I hope this has been of hel...

Read More

Thank you for your reply.

I will organise the peptide to be added to the catalogue for you. Once this has been done I will let you know.

If youhave any furtherquestions please do not hesitate to contact us.

Thank you for contacting us on Friday in regards to the immunogen used to raise the anti-PKC alpha antibody [Y124] (ab32376).

I am please do say that I am able to share some information regarding the immunogen sequence. I cannot give you the ...

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 17 2012

According to our records, ab32336 was proving difficult to use inWB and we were in contact in order to help resolve the issue. Looking at our correspondence, it appears that we are awaiting more details in order to help us better understand the difficu...

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Thank you for contacting us. I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly. I am atta...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Rat Tissue sections (Retina)
Specification Retina
Fixative Paraformaldehyde
Blocking step Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 24°C
Username

Abcam user community

Verified customer

Submitted Sep 12 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Human Tissue sections (Retina)
Specification Retina
Fixative Paraformaldehyde
Blocking step Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 24°C
Username

Abcam user community

Verified customer

Submitted Sep 12 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (human T84 cells)
Specification human T84 cells
Fixative Methanol
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C
Username

Dr. Jun Tang

Verified customer

Submitted Nov 06 2010

1-10 of 12 Abreviews or Q&A

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