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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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. {TM}

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
Class	Recombinant Polyclonal
Type	Antibody
Clone	2HCLC
Immunogen	Peptide corresponding to Human CHEK2 (aa 30-38) View antigen
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

Checkpoint kinase 2 (Chk2) is a serine/threonine-protein kinase which functions as a cell cycle checkpoint regulator. ATM and ATR kinases are activated in response to ionizing and ultraviolet radition, and phosphorylate Thr68 and other sites in the amino-terminal region of Chk2 dimerisation and activation. Activated Chk2 phosphorylates and inhibits Cdc25C phosphates, preventing the entry into mitosis, and has been shown to stabilize the tumor suppressor protein p53, leading to cell cycle arrest in G1. 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 this gene 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