Anti-CKII alpha 抗体 [8E5] - ChIP Grade (ab70774)

製品の概要

  • 製品名Anti-CKII alpha antibody [8E5] - ChIP Grade
    CKII alpha 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [8E5] to CKII alpha - ChIP Grade
  • アプリケーション適用あり: ICC/IF, ChIP, WB, IP, ELISA, Flow Cyt, IHC-Pmore details
  • 種交差性
    交差種: Mouse, Rat, Human, African Green Monkey
    交差が予測される動物種: Chicken, Cow, Xenopus laevis
  • 免疫原

    Recombinant human protein purified from E.coli (His-Casein kinase II a)

  • ポジティブ・コントロール
    • Jurkat T cell lysate

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab70774 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
ICC/IF 1/500. (see Abreview)
ChIP Use at an assay dependent concentration.
WB 1/2000. Predicted molecular weight: 45 kDa.
IP Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
Flow Cyt Use 0.5µg for 106 cells. ab170191-Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
IHC-P Use a concentration of 2 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

ターゲット情報

Anti-CKII alpha antibody [8E5] - ChIP Grade 画像



  • Predicted band size : 45 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: CKII alpha knockout HAP1 cell lysate (20 µg)
    Lane 3: HeLa cell lysate (20 µg)
    Lane 4: Jurkat cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab70774 observed at 44 kDa. Red - loading control, ab181602, observed at 37 kDa.
    ab70774 was shown to recognize CKII alpha when CKII alpha knockout samples were used, along with additional cross-reactive bands. Wild-type and CKII alpha knockout samples were subjected to SDS-PAGE. ab70774 and ab181602 (loading control to GAPDH) were diluted at 1/2000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • ab70774 (4µg/ml) staining CKII alpha in human cerebellum, using an automated system (DAKO Autostainer Plus). Using this protocol there is nuclear and cytoplasmic staining of purkinje cells.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
  • All lanes : Anti-CKII alpha antibody [8E5] - ChIP Grade (ab70774) at 1/2000 dilution

    Lane 1 : Jurkat T cell lysate
    Lane 2 : K562 cell lysate
    Lane 3 : NIH3T3 cell lysate
    Lane 4 : C6 cell lysate


    Predicted band size : 45 kDa
  • Overlay histogram showing Jurkat cells stained with ab70774 (red line). The cells were fixed with 80% methanol (5 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 (ab70774, 0.5µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 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 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

Anti-CKII alpha antibody [8E5] - ChIP Grade (ab70774) 使用論文

This product has been referenced in:
  • Basnet H  et al. Tyrosine phosphorylation of histone H2A by CK2 regulates transcriptional elongation. Nature N/A:N/A (2014). Read more (PubMed: 25252977) »
  • McFarland TP  et al. The cytosolic protein kinase CK2 phosphorylates cardiac calsequestrin in intact cells. Mol Cell Biochem 353:81-91 (2011). WB ; Human . Read more (PubMed: 21431367) »

See all 2 Publications for this product

Product Wall

Application Western blot
Sample Mouse Cell lysate - whole cell (Oocyte)
Gel Running Conditions Non-reduced Denaturing (12% SDS-PAGE gel)
Loading amount 200 cells
Specification Oocyte
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
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投稿 Jun 17 2015

Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing (10)
Sample African Green Monkey Cell lysate - whole cell (COS7)
Specification COS7
Blocking step (agent) for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 20°C
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投稿 Apr 28 2014

Abreviews
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Application Immunoprecipitation
Immuno-precipitation step Protein G
Sample Human Cell lysate - whole cell (HEK293)
Specification HEK293
Total protein in input 100 µg
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投稿 Jun 14 2013

Abreviews
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Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (10%)
Sample Mouse Cell lysate - whole cell (primary neuron)
Specification primary neuron
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 20°C
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投稿 Jun 05 2013

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Application Western blot
Sample Mouse Cell lysate - whole cell (Mouse T-cell lymphoma, macrophages and splenocytes)
Loading amount 40 µg
Specification Mouse T-cell lymphoma, macrophages and splenocytes
Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Dr. Tomar Ghansah

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投稿 Aug 21 2012

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Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (cervical tumor cell lines)
Specification cervical tumor cell lines
Fixative Paraformaldehyde
Permeabilization Yes - triton
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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投稿 Jan 14 2011

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Application Western blot
Sample Human Cell lysate - nuclear (keratinocytes)
Loading amount 50 µg
Specification keratinocytes
Gel Running Conditions Reduced Denaturing
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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投稿 Jan 04 2011

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Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Keratinocytes)
Specification Keratinocytes
Fixative Paraformaldehyde
Permeabilization Yes - 0.1% Triton-X-100
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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投稿 Jan 04 2011

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Application ChIP
Sample Human Cell lysate - nuclear (cervical tumor cells)
Specification cervical tumor cells
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: formaldehyde
Detection step Semiquantitative PCR
Positive control immortalized keratinocytes
Negative control HeLa tumor cells
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投稿 Dec 31 2010

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Application Western blot
Sample Human Cell lysate - nuclear (human keratinocytes and cervical cancer cells)
Loading amount 50 µg
Specification human keratinocytes and cervical cancer cells
Gel Running Conditions Reduced Denaturing (12%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Dr. Bladimiro Rincon Orozco

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投稿 Dec 02 2010

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