Overview

  • Product name

  • Description

    Rabbit polyclonal to CSNK2A2
  • Host species

    Rabbit
  • Specificity

    Reacts with the alpha prime subunit.
  • Tested applications

    Suitable for: IP, ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rabbit, Cow, Dog, Pig, Chimpanzee, Rhesus monkey, Gorilla, Orangutan, Bat, Elephant
  • Immunogen

    Immunogen was a synthetic peptide, which represented a portion of human Casein kinase 2, alpha 2 polypeptide encoded within exon 11 (LocusLink ID 1459).

  • Positive control

    • RIPA extract from HeLa cells.
  • General notes


    CKII is a serine/threonine protein kinase that phosphorylates acidic proteins such as casein. The kinase exists as a tetramer and is composed of an alpha, an alpha-prime, and two beta subunits. The alpha subunits contain the catalytic activity while the beta subunits undergo autophosphorylation. The gene for this protein is found on chromosome 20, but a related transcribed pseudogene is found on chromosome 11. Three transcript variants have been found, encoding two different isoforms. Protein kinase CKII is recognized as one of the key cellular signals for cell growth and proliferation; it also has an important role in apoptosis.

     This product was previously labelled as CKII alpha prime polypeptide

     

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    Preservative: 0.1% Sodium azide
    Constituents: 0.021% PBS, 1.764% Sodium citrate, 1.815% Tris
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Purification notes

    Antibodies were affinity purified using the peptide immobilized on solid support.
  • Primary antibody notes

    CKII is a serine/threonine protein kinase that phosphorylates acidic proteins such as casein. The kinase exists as a tetramer and is composed of an alpha, an alpha-prime, and two beta subunits. The alpha subunits contain the catalytic activity while the beta subunits undergo autophosphorylation. The gene for this protein is found on chromosome 20, but a related transcribed pseudogene is found on chromosome 11. Three transcript variants have been found, encoding two different isoforms. Protein kinase CKII is recognized as one of the key cellular signals for cell growth and proliferation; it also has an important role in apoptosis.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab10474 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
IP Use at 4-10 µg/mg of lysate.
ICC/IF Use at an assay dependent concentration.
WB 1/1000 - 1/5000. Detects a band of approximately 39 kDa (predicted molecular weight: 41 kDa).

Target

  • Function

    Catalytic subunit of a constitutively active serine/threonine-protein kinase complex that phosphorylates a large number of substrates containing acidic residues C-terminal to the phosphorylated serine or threonine. Regulates numerous cellular processes, such as cell cycle progression, apoptosis and transcription, as well as viral infection. May act as a regulatory node which integrates and coordinates numerous signals leading to an appropriate cellular response. During mitosis, functions as a component of the p53/TP53-dependent spindle assembly checkpoint (SAC) that maintains cyclin-B-CDK1 activity and G2 arrest in response to spindle damage. Also required for p53/TP53-mediated apoptosis, phosphorylating 'Ser-392' of p53/TP53 following UV irradiation. Can also negatively regulate apoptosis. Phosphorylates the caspases CASP9 and CASP2 and the apoptotic regulator NOL3. Phosphorylation protects CASP9 from cleavage and activation by CASP8, and inhibits the dimerization of CASP2 and activation of CASP8. Regulates transcription by direct phosphorylation of RNA polymerases I, II, III and IV. Also phosphorylates and regulates numerous transcription factors including NF-kappa-B, STAT1, CREB1, IRF1, IRF2, ATF1, SRF, MAX, JUN, FOS, MYC and MYB. Phosphorylates Hsp90 and its co-chaperones FKBP4 and CDC37, which is essential for chaperone function. Regulates Wnt signaling by phosphorylating CTNNB1 and the transcription factor LEF1. Acts as an ectokinase that phosphorylates several extracellular proteins. During viral infection, phosphorylates various proteins involved in the viral life cycles of EBV, HSV, HBV, HCV, HIV, CMV and HPV.
  • Sequence similarities

    Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. CK2 subfamily.
    Contains 1 protein kinase domain.
  • Information by UniProt
  • Database links

  • Alternative names

    • Casein kinase 2 alpha prime polypeptide antibody
    • Casein kinase II alpha' chain antibody
    • Casein kinase II subunit alpha' antibody
    • CK II alpha' antibody
    • CK II antibody
    • CK2A2 antibody
    • CKII antibody
    • CSK22_HUMAN antibody
    • CSNK2A1 antibody
    • CSNK2A2 antibody
    • FLJ43934 antibody
    see all

Images

  • Detection of human CSNK2A1 by western blot of immunoprecipitates.
    Samples: Whole cell lysate (0.5 or 1.0 mg per IP reaction; 20% of IP loaded) from HeLa cells prepared using NETN lysis buffer.
    Antibodies: Affinity purified rabbit anti-CSNK2A1 antibody A300-199A (lot A300-199A-5) used for IP at 6 µg per reaction. CSNK2A1 was also immunoprecipitated by rabbit anti-CSNK2A1 antibody A300-198A. For blotting immunoprecipitated CSNK2A1, A300-199A was used at 1 µg/ml.
    Detection: Chemiluminescence

  • Detection of human and mouse CSNK2A1 by western blot.
    Samples: Whole cell lysate (50 µg) from HeLa, HEK293T, Jurkat, mouse TCMK-1, and mouse NIH 3T3 cells prepared using NETN lysis buffer.
    Antibody: Affinity purified rabbit anti-CSNK2A1 antibody A300-199A (lot A300-199A-5) used for WB at 0.1 µg/ml.
    Detection: Chemiluminescence

  • Detection of human CSNK2A2 by Western blot and immunoprecipitation. 

    A. RIPA extract (30 mg) from HeLa cells treated with CSNK2A2 siRNA or vimentin siRNA (v).

    B. RIPA extract (30 mg) from HeLa cells treated with CSNK2A2, CSNK2A2 prime, or vimentin (v) siRNA.

    C. RIPA extract (0.5mg) from HeLa cells.

    Antibody: ab10474 used at 0.33 (A) or 0.2 (B) mg/ml for WB or 5 mg/0.5 mg lysate (C) for IP.

    Detection of human CSNK2A2 by Western blot and immunoprecipitation. A. RIPA extract (30 mg) from HeLa cells treated with CSNK2A2 siRNA or vimentin siRNA (v). B. RIPA extract (30 mg) from HeLa cells treated with CSNK2A2, CSNK2A2, or vimentin (v) siRNA. C. RIPA extract (0.5mg) from HeLa cells. Antibody: ab10474 used at 0.33 (A) or 0.2 (B) mg/ml for WB or 5 ug/0.5 mg lysate (C) for IP.

  • All lanes : Anti-CSNK2A2 antibody (ab10474) at 1/500 dilution

    Lane 1 : Whole cell lysate prepared from alpha prime knock out MEFs
    Lane 2 : Whole cell lysate prepared from alpha prime heterozygous MEFs

    Lysates/proteins at 40 µg per lane.

    Secondary
    All lanes : Goat anti rabbit IgG-HRP at 1/5000 dilution

    Developed using the ECL technique.

    Predicted band size: 41 kDa
    Observed band size: 37 kDa
    why is the actual band size different from the predicted?


    Exposure time: 30 minutes

    See Abreview

References

This product has been referenced in:

  • Beale AD  et al. Casein Kinase 1 Underlies Temperature Compensation of Circadian Rhythms in Human Red Blood Cells. J Biol Rhythms 34:144-153 (2019). Read more (PubMed: 30898060) »
  • Hoque A  et al. Mitochondrial fission protein Drp1 inhibition promotes cardiac mesodermal differentiation of human pluripotent stem cells. Cell Death Discov 4:39 (2018). WB . Read more (PubMed: 29531836) »
See all 5 Publications for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Human primary fibroblasts)
Permeabilization
Yes - 0.2% Triton-X100
Specification
Human primary fibroblasts
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: RT°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Dec 12 2013

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

Dr. Tomar Ghansah

Verified customer

Submitted Aug 21 2012

Answer

Thank you for your reply.

We only have the full size vials of all of our products, so unfortunatelywe don't have a small sample to send out.However, we will guarantee that this antibody is specific for CKII alpha prime and will work in the tested species and applications stated on the datasheet.

Please let me know if you have any further questions or if there is anything else that I can help you with. Have a great evening!

Read More

Question
Answer

Thank you for your call today and for your patience while I have been in touch with the lab about ab10474.

The immunogen of ab10474 was taken from a region of divergence between CKII alpha and CKII alpha', so the antibody is not cross-reactive. In the Western blot assays on the datasheet, no cross-reaction was seen between this antibody and CKII alpha.

I hope this information will be useful, but please let me know if you have any further questions and I'll be happy to help.

Read More
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse embryonic fibroblasts)
Gel Running Conditions
Reduced Denaturing (10% Tris-Gly gel)
Loading amount
20 µg
Specification
Mouse embryonic fibroblasts
Blocking step
LI-COR® Odyssey® Blocking Buffer as blocking agent for 45 minute(s) · Concentration: 50% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Nov 23 2011

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse Embryonic Fibroblasts (MEFs))
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
40 µg
Specification
Mouse Embryonic Fibroblasts (MEFs)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Jul 20 2011

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (HeLa cells)
Total protein in input
200 µg
Immuno-precipitation step
Protein A/G
Specification
HeLa cells

Abcam user community

Verified customer

Submitted Jan 18 2011

Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa cells)
Gel Running Conditions
Reduced Denaturing (10% Tris-Gly)
Loading amount
30 µg
Treatment
siRNA to CKII alpha prime
Specification
HeLa cells
Blocking step
LI-COR® Odyssey® Blocking Buffer as blocking agent for 45 minute(s) · Concentration: 50% · Temperature: RT°C

Dr. Svetlana Khoronenkova

Verified customer

Submitted Jul 22 2010

Answer

Thank you for your enquiry and patience. Under the conditions tested, which included WB detection at optimized dilutions using lysate from cells expressing endogenous amounts of the subunits, they were specific as specified. That is, ab10466, ab10468 and ab10470 react with the alpha subunit and ab10474 reacts with the alpha prime subunit. We would recommend loading 20-30 ug protein lysate. Please contact us again if you have any additional questions.

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