Invitrogen

Phospho-CHK2 (Ser33, Ser35) Recombinant Polyclonal Antibody (2HCLC)

Advanced Verification

This Antibody was verified by Cell treatment to ensure that the antibody binds to the antigen stated.

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Datasheet

Protocols

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Phospho-CHK2 (Ser33, Ser35) Antibody in Immunocytochemistry (ICC/IF) — For immunofluorescence analysis A549 cells were fixed and permeabilized for detection of Chk2 (pS33/35) using Chk2 (pS33/35) Recombinant Rabbit Polyclonal Antibody (Product # 710753, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Nuclei (blue) were stained using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938), and Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (red) staining. Detection and localization of Chk2 (pS33/35) (green) in the nucleus can be clearly observed in cells treated with UV (302 nm, 60 min) as compared to untreated cells. Antibody specificity was demonstrated by competition with the Chk2 (pS33/35) peptide, which results in loss of signal. No competition was observed with the non-phospho peptide.

Phospho-CHK2 (Ser33, Ser35) Antibody in Western Blot (WB) — Western blot analysis was performed on whole cell extracts (30 µg lysate) of A549 (Lane 1), A549 treated with Doxorubucin (1mM Doxorubicin for 2hrs) (Lane 2), A549 treated with UV (for 3hrs) (Lane 3), COLO 205 (Lane 4), and COLO 205 treated with Doxorubicin (1mM Doxorubicin for 2hrs) (Lane 5). The blots were probed with Anti-Chk2pS33/35 Recombinant Rabbit Polyclonal Antibody (Product # 710753, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 50 kDa band corresponding to Chk2pS33/35 was enhanced upon treatment. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10% Bis-Tris gel (Product # NP0301BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Phospho-CHK2 (Ser33, Ser35) Antibody — Altered expression of proteins upon cell treatment demonstrates antibody specificity. Immunofluorescence of Phospho-CHK2 (Ser33, Ser35) using Anti-Phospho-CHK2 (Ser33, Ser35) Recombinant Rabbit Polyclonal Antibody (Product # 710753), shows increased expression of Phospho-CHK2 (Ser33, Ser35) in A549 cells upon UV treatment. CHK2 is a DNA damage signal transducer kinase that is activated by ATM upon UV-induced DNA damage.

Product Details

710753

Applications

Tested Dilution

Publications

Western Blot (WB)

1-2 µg/mL

Immunocytochemistry (ICC/IF)

2 µg/mL

Product Specifications

Species Reactivity

Human

Host/Isotype

Rabbit / IgG

Expression System

EXPI293

Class

Recombinant Polyclonal

Type

Antibody

Clone

2HCLC

Immunogen

Peptide corresponding to Human CHEK2 (aa 30-38)

View immunogen

Conjugate

Unconjugated

Form

Liquid

Concentration

0.5 mg/mL

Purification

Protein A

Storage buffer

PBS, pH 7.2

Contains

0.09% sodium azide

Storage conditions

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

RRID

AB_2609851

Product Specific Information

This antibody is predicted to react with Monkey, Pig and Rabbit.

Recombinant rabbit polyclonal antibodies are unique offerings from Thermo Fisher Scientific. They are comprised of a selection of multiple different recombinant monoclonal antibodies, providing the best of both worlds - the sensitivity of polyclonal antibodies with the specificity of monoclonal antibodies - all delivered with the consistency only found in a recombinant antibody. While functionally the same as a polyclonal antibody - recognizing multiple epitope sites on the target and producing higher detection sensitivity for low abundance targets - a recombinant rabbit polyclonal antibody has a known mixture of light and heavy chains. The exact population can be produced in every lot, circumventing the biological variability typically associated with polyclonal antibody production.

Target Information

CHEK2 (CHK2) is a serine/threonine kinase and a component of the DNA damage checkpoint pathway. A mutation in CHK2 has been linked to cancer. CHK2 is activated by DNA damage and phosphorylates several modulators of cell cycle control including tumor suppressor proteins. In addition, Chk2 can phosphorylate BRCA1, allowing BRCA1 to restore survival after DNA damage. Chk2 is a putative tumor suppressor protein, with mutations in to the Chk2 gene linked to Li-Fraumeni syndrome, a familiar cancer usually associated with inherited mutations in TP53. Also, mutations in CHK2 are thought to confer a predisposition to sarcomas, breast cancer, and brain tumors.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: Cds1 homolog; Checkpoint kinase 2; Checkpoint like protein CHK2; checkpoint-like protein CHK2; Chk2 (phospho T68); Chk2 (pThr68); CHK2 checkpoint homolog; CHK2 checkpoint homolog (S. pombe); Chk2 phospho; Chk2 phospho T68; del9; EC 2.7.11.1; HuCds 1; Hucds1; kinase Chk2; OTTHUMP00000199044; OTTHUMP00000199045; OTTHUMP00000199064; OTTHUMP00000199115; OTTHUMP00000199116; phospho Chk2 (T68); phospho Thr68 Chk2; RP11-436C9.1; Serine/threonine protein kinase Chk2; Serine/threonine-protein kinase Chk2

Gene Aliases: CDS1; CHEK2; CHK2; hCds1; HuCds1; LFS2; PP1425; RAD53

UniProt ID: (Human) O96017

Entrez Gene ID: (Human) 11200

Molecular Function: non-receptor serine/threonine protein kinase