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

Overview

  • Product name
    Anti-CKII alpha antibody [8E5] - ChIP Grade
    See all CKII alpha primary antibodies
  • Description
    Mouse monoclonal [8E5] to CKII alpha - ChIP Grade
  • Tested applications
    Suitable for: ICC/IF, ChIP, WB, IP, ELISA, Flow Cyt, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, African green monkey
    Predicted to work with: Chicken, Cow, Xenopus laevis
  • Immunogen

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

  • Positive control
    • Jurkat T cell lysate
  • General notes

    This product was changed from ascites to tissue culture supernatant on 18th September 2017. Lot numbers higher than GR314900 will be from tissue culture supernatant. Please note that the dilutions may need to be adjusted accordingly.

Properties

Applications

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.

Application Abreviews Notes
ICC/IF Use at an assay dependent concentration.

(see Abreview)

ChIP Use at an assay dependent concentration.
WB Use a concentration of 0.5 µg/ml. Predicted molecular weight: 45 kDa.
IP Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab170191-Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

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



  • 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-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) 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.

References for Anti-CKII alpha antibody [8E5] - ChIP Grade (ab70774)

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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Submitted 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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Submitted Apr 28 2014

Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
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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Submitted Jun 14 2013

Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
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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Submitted Jun 05 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Dr. Tomar Ghansah

Verified customer

Submitted Aug 21 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
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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Submitted Jan 14 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
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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Verified customer

Submitted Jan 04 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
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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Submitted Jan 04 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
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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Submitted Dec 31 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Dr. Bladimiro Rincon Orozco

Verified customer

Submitted Dec 02 2010

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

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