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Signal Transduction Protein Phosphorylation Ser / Thr Kinases PKC
使用敲除细胞株进行验证重组RabMAb

重组Anti-PKC alpha抗体[Y124] (ab32376)

  • Datasheet
  • SDS
Reviews (13)Q&A (5)References (57)

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Western blot - Anti-PKC alpha antibody [Y124] (ab32376)
  • Immunocytochemistry/ Immunofluorescence - Anti-PKC alpha antibody [Y124] (ab32376)
  • Immunoprecipitation - Anti-PKC alpha antibody [Y124] (ab32376)
  • Immunoprecipitation - Anti-PKC alpha antibody [Y124] (ab32376)
  • Immunocytochemistry/ Immunofluorescence - Anti-PKC alpha antibody [Y124] (ab32376)
  • Western blot - Anti-PKC alpha antibody [Y124] (ab32376)
  • Flow Cytometry - Anti-PKC alpha antibody [Y124] (ab32376)
  • Immunocytochemistry/ Immunofluorescence - Anti-PKC alpha antibody [Y124] (ab32376)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha antibody [Y124] (ab32376)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha antibody [Y124] (ab32376)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha antibody [Y124] (ab32376)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha antibody [Y124] (ab32376)
  • Western blot - Anti-PKC alpha antibody [Y124] (ab32376)
  • Western blot - Anti-PKC alpha antibody [Y124] (ab32376)
  • Flow Cytometry - Anti-PKC alpha antibody [Y124] (ab32376)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha antibody [Y124] (ab32376)
  • Western blot - Anti-PKC alpha antibody [Y124] (ab32376)
  • Immunocytochemistry/ Immunofluorescence - Anti-PKC alpha antibody [Y124] (ab32376)
  • Anti-PKC alpha antibody [Y124] (ab32376)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y124] to PKC alpha
  • Suitable for: WB, IHC-P, Flow Cyt, IP, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human, Pig

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概述

  • 产品名称

    Anti-PKC alpha抗体[Y124]
    参阅全部 PKC alpha 一抗
  • 描述

    兔单克隆抗体[Y124] to PKC alpha
  • 宿主

    Rabbit
  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Mouse
    Rat
    Human
    IP
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data
  • 免疫原

    Synthetic peptide within Human PKC alpha aa 650 to the C-terminus (C terminal). The exact sequence is proprietary.

  • 阳性对照

    • WB: HEK293, K562, HeLa, C6 cell lysates and pig skeletal muscle tissue lysates. IHC-P: Human lung carcinoma and human endometrium carcinoma tissues. ICC/IF: PMA-Treated and untreated wild-type HAP1 and HeLa cells.
  • 常规说明

    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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

性能

  • 形式

    Liquid
  • 存放说明

    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.
  • 存储溶液

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • 纯度

    Protein A purified
  • 克隆

    单克隆
  • 克隆编号

    Y124
  • 同种型

    IgG
  • 研究领域

    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • PKC
    • Cancer
    • Signal transduction
    • Protein phosphorylation
    • Serine/threonine kinases
    • Other
    • Cardiovascular
    • Heart
    • Hypertrophy
    • Other

相关产品

  • Alternative Versions

    • Alexa Fluor® 488 Anti-PKC alpha antibody [Y124] (ab204633)
    • Alexa Fluor® 647 Anti-PKC alpha antibody [Y124] (ab204634)
    • HRP Anti-PKC alpha antibody [Y124] (ab205422)
    • Alexa Fluor® 594 Anti-PKC alpha antibody [Y124] (ab206344)
    • Alexa Fluor® 405 Anti-PKC alpha antibody [Y124] (ab206345)
    • PE Anti-PKC alpha antibody [Y124] (ab208749)
    • Anti-PKC alpha antibody [Y124] - BSA and Azide free (ab221611)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HEK-293 whole cell lysate (ab7902)
  • Recombinant Protein

    • Recombinant human PKC alpha protein (Active) (ab55672)
  • Related Products

    • Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)

应用

The Abpromise guarantee

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

应用 Species
Flow Cyt
Human
ICC/IF
Human
IHC-P
Mouse
Rat
Human
IP
Human
WB
Mouse
Rat
Human
All applications
Goldfish
应用 Ab评论 说明
WB (6)
1/1000 - 1/10000. Detects a band of approximately 75 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.

For unpurified use at 1/250 - 1/500. 

Flow Cyt
1/20.

For unpurified use at 1/100. 

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

IP
1/20.

For unpurified use at 1/100.

ICC/IF (2)
1/200.

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

说明
WB
1/1000 - 1/10000. Detects a band of approximately 75 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.

For unpurified use at 1/250 - 1/500. 

Flow Cyt
1/20.

For unpurified use at 1/100. 

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

IP
1/20.

For unpurified use at 1/100.

ICC/IF
1/200.

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

靶标

  • 功能

    This 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.
  • 序列相似性

    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.
  • 细胞定位

    Cytoplasm. Cell membrane. Nucleus.
  • Target information above from: UniProt accession P17252 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 数据库链接

    • Entrez Gene: 5578 Human
    • Entrez Gene: 18750 Mouse
    • Entrez Gene: 397184 Pig
    • Entrez Gene: 24680 Rat
    • Omim: 176960 Human
    • SwissProt: P17252 Human
    • SwissProt: P20444 Mouse
    • SwissProt: P05696 Rat
    • Unigene: 531704 Human
    • Unigene: 708867 Human
    • Unigene: 222178 Mouse
    • Unigene: 207908 Rat
    see all
  • 别名

    • 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

图片

  • Western blot - Anti-PKC alpha antibody [Y124] (ab32376)
    Western blot - Anti-PKC alpha antibody [Y124] (ab32376)

    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 in wild-type HAP1 cells. No band was observed when PKC alpha knockout samples were examined. 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) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-PKC alpha antibody [Y124] (ab32376)
    Immunocytochemistry/ Immunofluorescence - Anti-PKC alpha antibody [Y124] (ab32376)

    Unpurified 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).

  • Immunoprecipitation - Anti-PKC alpha antibody [Y124] (ab32376)
    Immunoprecipitation - Anti-PKC alpha antibody [Y124] (ab32376)

    ab32376 (purified) at 1:20 dilution (0.5ug) immunoprecipitating PKC alpha in 293 (Human embryonic kidney epithelial cell) whole cell lysate.

    Lane 1 (input): 293 (Human embryonic kidney epithelial cell) whole cell lysate 10ug

    Lane 2 (+): ab32376 & 293 (Human embryonic kidney epithelial cell) whole cell lysate

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32376 in 293 (Human embryonic kidney epithelial cell) whole cell lysate

    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.

    Blocking and diluting buffer: 5% NFDM/TBST.

    The band between 37kDa and 50kDa might be the C-term fragment. (PMID:10381525)

  • Immunoprecipitation - Anti-PKC alpha antibody [Y124] (ab32376)
    Immunoprecipitation - Anti-PKC alpha antibody [Y124] (ab32376)

    ab32376 (purified) at 1:20 dilution (0.5ug) immunoprecipitating PKC alpha in Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate.

    Lane 1 (input): Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10ug

    Lane 2 (+): ab32376 & Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32376 in Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate


    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.

    Blocking and diluting buffer: 5% NFDM/TBST.

    The band between 37kDa and 50kDa might be the C-term fragment. (PMID:10381525)

  • Immunocytochemistry/ Immunofluorescence - Anti-PKC alpha antibody [Y124] (ab32376)
    Immunocytochemistry/ Immunofluorescence - Anti-PKC alpha antibody [Y124] (ab32376)

    Unpurified 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).

  • Western blot - Anti-PKC alpha antibody [Y124] (ab32376)
    Western blot - Anti-PKC alpha antibody [Y124] (ab32376)

    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 - unpurified ab32376 observed at 77 kDa. Red - loading control, ab8245, observed at 37 kDa.

    This western blot image is a comparison between ab32376 and a competitor's top cited rabbit polyclonal antibody.

  • Flow Cytometry - Anti-PKC alpha antibody [Y124] (ab32376)
    Flow Cytometry - Anti-PKC alpha antibody [Y124] (ab32376)

    Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling PKC alpha with purified ab32376 at 1:20 dilution (5 ug/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - 90% methanol. Unlabeled control - Rabbit monoclonal IgG (Black). Cell without incubation with primary antibody and secondary antibody (Blue).

  • Immunocytochemistry/ Immunofluorescence - Anti-PKC alpha antibody [Y124] (ab32376)
    Immunocytochemistry/ Immunofluorescence - Anti-PKC alpha antibody [Y124] (ab32376)

    Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling PKC alpha with Purified ab32376 at 1:250 dilution (0.4μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha antibody [Y124] (ab32376)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha antibody [Y124] (ab32376)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat liver tissue sections labeling PKC alpha with purified  ab32376 at 1:100 dilution (1.01 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using EDTA Buffer, pH9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha antibody [Y124] (ab32376)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha antibody [Y124] (ab32376)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human glioma tissue sections labeling PKC alpha with purified  ab32376 at 1:100 dilution (1.01 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using EDTA Buffer, pH9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha antibody [Y124] (ab32376)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha antibody [Y124] (ab32376)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse liver tissue sections labeling PKC alpha with purified  ab32376 at 1:100 dilution (1.01 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using EDTA Buffer, pH9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha antibody [Y124] (ab32376)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha antibody [Y124] (ab32376)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human endometrium carcinoma tissue sections labeling PKC alpha with purified ab32376 at 1:100 dilution (1.01 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using EDTA Buffer, pH9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

  • Western blot - Anti-PKC alpha antibody [Y124] (ab32376)
    Western blot - Anti-PKC alpha antibody [Y124] (ab32376)
    All lanes : Anti-PKC alpha antibody [Y124] (ab32376) at 1/5000 dilution (purified)

    Lane 1 : 293 (Human embryonic kidney epithelial cell) whole cell lysates
    Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates

    Lysates/proteins at 15 µg per lane.

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

    Predicted band size: 77 kDa



    Blocking and diluting buffer: 5% NFDM/TBST

  • Western blot - Anti-PKC alpha antibody [Y124] (ab32376)
    Western blot - Anti-PKC alpha antibody [Y124] (ab32376)
    All lanes : Anti-PKC alpha antibody [Y124] (ab32376) at 1/5000 dilution (purified)

    Lane 1 : C6 (Rat glial tumor glial cell) whole cell lysates
    Lane 2 : Pig skeletal muscle lysates

    Lysates/proteins at 15 µg per lane.

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

    Predicted band size: 77 kDa



    Blocking and diluting buffer: 5% NFDM/TBST

  • Flow Cytometry - Anti-PKC alpha antibody [Y124] (ab32376)
    Flow Cytometry - Anti-PKC alpha antibody [Y124] (ab32376)

    Overlay histogram showing HeLa cells stained with unpurified 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha antibody [Y124] (ab32376)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha antibody [Y124] (ab32376)Image from Hao F et al., J Biol Chem. 2011 May 20;286(20):18104-17. Epub 2011 Mar 18. Fig 1.; doi: 10.1074/jbc.M110.208488; May 20, 2011 The Journal of Biological Chemistry, 286, 18104-18117.

    Immunohistochemical analysis of mouse small intestine tissue, staining PKC alpha with unpurified ab32376.

  • Western blot - Anti-PKC alpha antibody [Y124] (ab32376)
    Western blot - Anti-PKC alpha antibody [Y124] (ab32376)
    All lanes : Anti-PKC alpha antibody [Y124] (ab32376) at 1/10000 dilution (purified)

    Lane 1 : K562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates
    Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 77 kDa
    Observed band size: 80 kDa why is the actual band size different from the predicted?



    Blocking and diluting buffer: 5% NFDM/TBST 
    The band between 37kDa and 50kDa might be the C-term fragment. (PMID:10381525)

  • Immunocytochemistry/ Immunofluorescence - Anti-PKC alpha antibody [Y124] (ab32376)
    Immunocytochemistry/ Immunofluorescence - Anti-PKC alpha antibody [Y124] (ab32376)This image is courtesy of an anonymous Abreview

    ab32376 staining PKC in the HT1080 Cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody (1/400 in PBS) for 1 hour at 22°C. An Alexa Fluor® 488-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody (1/1000).

    See Abreview

  • Anti-PKC alpha antibody [Y124] (ab32376)
    Anti-PKC alpha antibody [Y124] (ab32376)

实验方案

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

数据表及文件

  • SDS download

  • Datasheet download

    Download

文献 (57)

发表研究结果有使用 ab32376?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab32376 被引用在 57 文献中.

  • Yin C  et al. Two Types of Cl Transporters Contribute to the Regulation of Intracellular Cl Concentrations in ON- and OFF-type Bipolar Cells in the Mouse Retina. Neuroscience 440:267-276 (2020). PubMed: 32531472
  • Pi H  et al. AKT inhibition-mediated dephosphorylation of TFE3 promotes overactive autophagy independent of MTORC1 in cadmium-exposed bone mesenchymal stem cells. Autophagy 15:565-582 (2019). PubMed: 30324847
  • Li G  et al. Generation and Characterization of Induced Pluripotent Stem Cells and Retinal Organoids From a Leber's Congenital Amaurosis Patient With Novel RPE65 Mutations. Front Mol Neurosci 12:212 (2019). PubMed: 31572124
  • Pan X  et al. Essential Role Of High Glucose-Induced Overexpression Of PKCß And PKCd In GLP-1 Resistance In Rodent Cardiomyocytes. Diabetes Metab Syndr Obes 12:2289-2302 (2019). PubMed: 31807042
  • Alsaeedi HA  et al. Dental pulp stem cells therapy overcome photoreceptor cell death and protects the retina in a rat model of sodium iodate-induced retinal degeneration. J Photochem Photobiol B 198:111561 (2019). PubMed: 31352000
View all Publications for this product

客户评价及客户问答

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1-10 of 18 Abreviews or Q&A

IHC - Wholemount abreview for Anti-PKC alpha antibody [Y124]

Below Average
Abreviews
Abreviews
abreview image
Application
IHC - Wholemount
Sample
Mouse Embryo (E12.5 Mouse Embryo)
Specification
E12.5 Mouse Embryo
Read More

Dr. Alain Chedotal

Verified customer

提交于 Feb 25 2021

Western blot abreview for Anti-PKC alpha antibody [Y124]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Gastric Cancer Cell Line)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
3 µg
Specification
Gastric Cancer Cell Line
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Read More

Abcam user community

Verified customer

提交于 Nov 18 2020

Western blot abreview for Anti-PKC alpha antibody [Y124]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Macrophage)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
3 µg
Specification
Macrophage
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Read More

Abcam user community

Verified customer

提交于 Nov 13 2020

Western blot abreview for Anti-PKC alpha antibody [Y124]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Human umbilical vein endothelial cell)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (10%)
Loading amount
20 µg
Specification
Human umbilical vein endothelial cell
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

提交于 Nov 07 2017

Immunohistochemistry (Frozen sections) abreview for Anti-PKC alpha antibody [Y124]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Zebrafish Cell (5 um frozen retinal section)
Permeabilization
No
Specification
5 um frozen retinal section
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 20% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More

Dr. Jenny Lenkowski

Verified customer

提交于 Aug 02 2017

Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-PKC alpha antibody [Y124]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Fukomys anselli Tissue sections (Retina)
Antigen retrieval step
None
Permeabilization
No
Specification
Retina
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 21°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

提交于 Dec 21 2016

Western blot abreview for Anti-PKC alpha antibody [Y124]

Excellent
Abreviews
Abreviews
abreview image
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
Read More

Mr. Baptiste Jude

Verified customer

提交于 Jul 07 2016

Question

Thanks, that's helpful. Please add the blocking peptide to the catalogue.

Read More

Abcam community

Verified customer

Asked on Jun 06 2012

Answer

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 help. If you have any further questions please do not hesitate to contact us again.

Read More

Abcam Scientific Support

回复于 Jun 06 2012

Question

Thanks xxxxx, that's helpful. Please add the blocking peptide to the catalogue.

Read More

Abcam community

Verified customer

Asked on May 31 2012

Answer

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.

Read More

Abcam Scientific Support

回复于 May 31 2012

Question

Phone call requesting information of the immunogen used to raise anti-PKC alpha antibody [Y124] (ab32376).

Read More

Abcam community

Verified customer

Asked on May 28 2012

Answer

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 exact sequence used as this iscommercially sensitivebut I can say that theimmunogen used is a synthetic peptide taken from the C-terminus of human PKCa withinresidues 648 - 672 of human PKCa (UniProt: P17252).

I can arrange for the blocking peptide to be added to the catalogue for you if you would be interested.

I hope this information has been of help. If you require any further information please do not hesitate to contact us again.

Read More

Abcam Scientific Support

回复于 May 28 2012

1-10 of 18 Abreviews or Q&A

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