- PA5-34625 - Invitrogen Antibodies CHEK1 antibody

Submitted references The Ataxia telangiectasia-mutated and Rad3-related protein kinase regulates cellular hydrogen sulfide concentrations.

Chen J, Shen X, Pardue S, Meram AT, Rajendran S, Ghali GE, Kevil CG, Shackelford RE
DNA repair 2019 Jan;73:55-63

The ATM and Rad3-Related (ATR) Protein Kinase Pathway Is Activated by Herpes Simplex Virus 1 and Required for Efficient Viral Replication.

Edwards TG, Bloom DC, Fisher C
Journal of virology 2018 Mar 15;92(6)

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using Phospho-CHK1 (Ser345) Polyclonal Antibody (Product # PA5-34625). Sample (30 µg of whole cell lysate). Lane A: HCT116 cells with mock treatment for 24 hr. Lane B: HCT116 cells with 30 µM cisplatin treatment for 24 hr. 10% SDS PAGE. Phospho-CHK1 (Ser345) Polyclonal Antibody (Product # PA5-34625) diluted at 1:500. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using Phospho-CHK1 (Ser345) Polyclonal Antibody (Product # PA5-34625). Untreated (–) and treated (+) HCT116 whole cell extracts (30 µg) were separated by 10% SDS-PAGE, and the membrane was blotted with Phospho-CHK1 (Ser345) Polyclonal Antibody (Product # PA5-34625) diluted at 1:500. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody, and the signal was developed with Trident ECL plus-Enhanced.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-Phospho-CHK1 (Ser345) Polyclonal Antibody (Product # PA5-34625) and a 55 kDa band corresponding to Phospho-CHK1 (Ser345) was observed to be induced upon Calyculin A, Etoposide and Hydroxyurea treatments in HeLa cells and Calyculin A treatment in U-2 OS cells. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), HeLa treated with Calyculin A (100 nM for 1 hour) (Lane 2), HeLa (Lane 3), HeLa treated with Etoposide (5 µM for 16 hours) (Lane 4), U-2 OS (Lane 5), U-2 OS treated with Calyculin A (100 nM for 1 hour) (Lane 6), HeLa (Lane 7) and HeLa treated with Hydroxyurea (10 mM for 2 hours) (Lane 8) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0301BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:20,000 dilution) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Atto Ultimate Sensitivity Substrate (Product # A38556). An uncharacterized band of ~38 kDa was also observed across the cell lines tested and ~110 kDa band was observed in HeLa cells treated with Etoposide.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using Phospho-CHK1 (Ser345) Polyclonal Antibody (Product # PA5-34625). Sample: 20 µg of HCT116 P53 whole cell lysate 10% SDS PAGE. Chk1-phopho-S345 antibody. Phospho-CHK1 (Ser345) Polyclonal Antibody (Product # PA5-34625) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on modified whole cell lysate (1% SDS) (30 µg lysate) of HeLa (Lane 1) and HeLa treated with Etoposide (5uM, 16 hrs) (Lane 2). The blot was probed with Anti- pCHK1 (Ser345) Polyclonal Antibody (Product # PA5-34625, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). A 54 kDa band corresponding to pCHK1 (Ser345) was observed in Hela cells upon etoposide treatment.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry-Immunofluorescence analysis of Phospho-CHK1 (Ser345) using Phospho-CHK1 (Ser345) Polyclonal Antibody (Product # PA5-34625) (Green) diluted at 1:500. HeLa cells mock (left) and treated with 100 J/m2 UVC and recover for 8 hrs (right) were fixed in 4% paraformaldehyde at RT for 15 min. Blue: Hoechst 33342 staining. Scale bar = 10 µm.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Phospho-CHK1 (Ser345) Polyclonal Antibody detects Chk1 (phospho Ser345) protein at nucleus by immunohistochemical analysis. Sample: Paraffin-embedded mouse intestine. Chk1 (phospho Ser345) stained by Phospho-CHK1 (Ser345) Polyclonal Antibody (Product # PA5-34625) diluted at 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Phospho-CHK1 (Ser345) Polyclonal Antibody detects Chk1 (phospho Ser345) protein at nucleus on mouse colon by immunohistochemical analysis. Sample: Paraffin-embedded mouse colon. Phospho-CHK1 (Ser345) Polyclonal Antibody (Product # PA5-34625) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of paraffin-embedded HeLa xenograft, using Chk1 (phospho Ser345) (Product # PA5-34625) antibody at 1:500 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

PA5-34625

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