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ImmunohistochemistryAntibody data
- Antibody Data
- Antigen structure
- References [2]
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- Validations
- Western blot [5]
- Immunocytochemistry [1]
- Other assay [2]
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- Product number
- PA5-34625 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Phospho-CHK1 (Ser345) Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- Recommended positive controls: HCT116, HCT116 cells with 30uM cisplatin treatment for 24hr. Predicted reactivity: Mouse (100%), Rat (100%), Bovine (100%). IHC notes, Requires antigen retrieval using heat mediated 10mM Citrate buffer (pH6.0) or Tris-EDTA buffer (pH8.0) Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
- Reactivity
- Human, Mouse
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 1.66 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
Submitted references The Ataxia telangiectasia-mutated and Rad3-related protein kinase regulates cellular hydrogen sulfide concentrations.
The ATM and Rad3-Related (ATR) Protein Kinase Pathway Is Activated by Herpes Simplex Virus 1 and Required for Efficient Viral Replication.
Chen J, Shen X, Pardue S, Meram AT, Rajendran S, Ghali GE, Kevil CG, Shackelford RE
DNA repair 2019 Jan;73:55-63
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)
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.
- 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.
- 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.
- 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.
- 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
- NULL
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- NULL
