Immunofluorescent analysis of Phospho-CHK1 pSer345 showing staining in the nucleus of HeLa cells. HeLa cells mock (left) and UVC treated with 100J/m2 and recover for 8hr (right) were fixed in 4% paraformaldehyde at RT for 15 min and stained using a Phospho-CHK1 pSer345 polyclonal antibody (Product # PA5-34625) diluted at 1:500. Blue: Hoechst 33342 staining.
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic phospho-peptide corresponding to amino acids residues surrounding the phospho Ser345 of CHK1|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS, pH 7, with 1% BSA, 20% glycerol|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:100-1:1000|
|Western Blot (WB)||1:500-1:3000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
IHC notes, Requires antigen retrieval using heat mediated 10mM Citrate buffer (pH6.0) or Tris-EDTA buffer (pH8.0)
Cell cycle events are regulated by the sequential activation and deactivation of cyclin dependent kinases (Cdks) and by proteolysis of cyclins. Chk1 and Chk2 are involved in these processes as regulators of Cdks. Chk1 and Chk2 both function as essential components in the G2 DNA damage checkpoint by phosphorylating Cdc25C in response to DNA damage. Phosphorylation inhibits Cdc25C activity, thereby blocking mitosis. Cdc25A, Cdc25B and Cdc25C protein tyrosine phosphatases function as mitotic activators by dephosphorylating Cdc2 p34 on regulatory tyrosine residues. It has also been shown that Chk1 can phosphorylate Wee 1 in vitro, providing evidence that the hyperphosphorylated form of Wee 1, seen in cells delayed by Chk1 overexpression, is due to phosphorylation by Chk1. Chk1 is phosphorylated on Serine 345 (S345) in response to UV, IR and hydroxyurea (HU). Chk1 plays an essential role in the mammalian DNA damage checkpoint, embryonic development and tumor suppression.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.