|Validated Species:||Human, Mouse, Rat|
Data & Images
|Product Name||Anti-Chk2 Antibody|
|Description||Rabbit IgG polyclonal antibody for Serine/threonine-protein kinase Chk2(CHEK2) detection. Tested with WB in Human;Mouse;Rat.|
|Cite This Product||Anti-Chk2 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1202)|
|Replacement Item||This antibody may replace the following items: sc-101658|sc-136251|sc-16297-R|sc-17747|sc-17748|sc-5278|sc-56296|sc-56297|sc-8812|sc-8813|sc-8813-R|sc-9064 from Santa Cruz Biotechnology.|
|Validated Species||Human, Mouse, Rat|
*This antibody is predicted to react with the above species based on antigen sequence similarities. Our Boster Guarantee covers the use of this product with the above species.
*Our Boster Guarantee covers the use of this product in the above tested applications.
**For positive and negative control design, consult "Tissue specificity" under Protein Target Info.
|Recommended Detection Systems||Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot.
*Blocking peptide can be purchased at $50. Contact us for more information
**Boster also offers various secondary antibodies for Immunoflourescecne and IHC. Take advantage of the buy 1 primary antibody get 1 secondary antibody for free promotion for the entire year 2018!
|Immunogen||A synthetic peptide corresponding to a sequence at the C-terminus of human Chk2(427-441aa FSEHRTQVSLKDQIT), different from the related rat and mouse sequences by one amino acid.|
|Cross Reactivity||No cross reactivity with other proteins|
|Contents||Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.|
|Concentration||Add 0.2ml of distilled water will yield a concentration of 500ug/ml.|
|Storage||At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.|
|Purification||Immunogen affinity purified.|
Protein Target Info (Source: Uniprot.org)
You can check the tissue specificity below for information on selecting positive and negative control.
|Protein Name||Serine/threonine-protein kinase Chk2|
|Molecular Weight||60915 MW|
|Protein Function||Serine/threonine-protein kinase which is required for checkpoint-mediated cell cycle arrest, activation of DNA repair and apoptosis in response to the presence of DNA double-strand breaks. May also negatively regulate cell cycle progression during unperturbed cell cycles. Following activation, phosphorylates numerous effectors preferentially at the consensus sequence [L-X- R-X-X-S/T]. Regulates cell cycle checkpoint arrest through phosphorylation of CDC25A, CDC25B and CDC25C, inhibiting their activity. Inhibition of CDC25 phosphatase activity leads to increased inhibitory tyrosine phosphorylation of CDK-cyclin complexes and blocks cell cycle progression. May also phosphorylate NEK6 which is involved in G2/M cell cycle arrest. Regulates DNA repair through phosphorylation of BRCA2, enhancing the association of RAD51 with chromatin which promotes DNA repair by homologous recombination. Also stimulates the transcription of genes involved in DNA repair (including BRCA2) through the phosphorylation and activation of the transcription factor FOXM1. Regulates apoptosis through the phosphorylation of p53/TP53, MDM4 and PML. Phosphorylation of p53/TP53 at 'Ser-20' by CHEK2 may alleviate inhibition by MDM2, leading to accumulation of active p53/TP53. Phosphorylation of MDM4 may also reduce degradation of p53/TP53. Also controls the transcription of pro-apoptotic genes through phosphorylation of the transcription factor E2F1. Tumor suppressor, it may also have a DNA damage-independent function in mitotic spindle assembly by phosphorylating BRCA1. Its absence may be a cause of the chromosomal instability observed in some cancer cells. Promotes the CCAR2-SIRT1 association and is required for CCAR2-mediated SIRT1 inhibition (PubMed:25361978). .|
|Tissue Specificity||High expression is found in testis, spleen, colon and peripheral blood leukocytes. Low expression is found in other tissues.|
|Sequence Similarities||Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. CHK2 subfamily.|
|Subcellular Localization||Isoform 2: Nucleus. Isoform 10 is present throughout the cell.|
|Alternative Names||Serine/threonine-protein kinase Chk2;18.104.22.168;CHK2 checkpoint homolog;Cds1 homolog;Hucds1;hCds1;Checkpoint kinase 2;CHEK2;CDS1, CHK2, RAD53;|
|Research Areas|||epigenetics and nuclear signaling|dna / rna|dna damage & repair|dna damage response|chk1 / chk2| cancer|cell cycle|kinases/phosphatases||
Background for Serine/threonine-protein kinase Chk2
Dilution Ratios/Recommended Concentrations
At Boster we strive to provide the best Anti-Chk2 Antibody by testing all applications on non-spiked tissues and cell lines to ensure that the affinity of the antibody is enough to react to the endogenouse level of the target protein. Read more about our QC panel here.
|Recommended dilution ratios are listed below:|
Western blot, 0.1-0.5μg/ml, Human, Rat, Mouse|
**Boster provides high sensitivity secondary antibody kits for Western blotting and IHC. For more info see Related Products below.
Anti-Chk2 Antibody Images
Click the images to enlarge.
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human MDA-MB-231 whole cell lysates,
Lane 2: human MDA-MB-453 whole cell lysates,
Lane 3: human HepG2 whole cell lysates,
Lane 4: human K562 whole cell lysates,
Lane 5: human Caco-2 whole cell lysates,
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti- Chk2 antigen affinity purified polyclonal antibody (Catalog # PA1202) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Chk2 at approximately 69KD. The expected band size for Chk2 is at 61KD.
1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected
2. Post-translational cleavage: this can cause smaller bands and or multiple bands
3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody.
4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.
5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher.,