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Phospho-Chk1 (S345) Recombinant Rabbit Monoclonal Antibody [PS01-17]

Usd: 385

Specification

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Catalog# HA721189

Phospho-Chk1 (S345) Recombinant Rabbit Monoclonal Antibody [PS01-17]

Application

  • WB

  • IF-Cell

Reactivity

  • Human

  • Mouse

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Phospho-Chk1 (S345) Recombinant Rabbit Monoclonal Antibody [PS01-17]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic phosphopeptide corresponding to residues surrounding Ser345 of human Chk1.

Species Reactivity

Human, Mouse

Validated Applications

WB, IF-Cell

Molecular Weight

Predicted band size: 54 kDa

Positive Control

HEK-293 treated with 200nM Calyculin A for 1 hour cell lysate, NIH/3T3 treated with UV for 20 minutes then recover for 2 hours cell lysate, HeLa treated with Etoposide cell lysate, HeLa treated with Hydroxyurea cell lysate, HeLa treated with 4mM Hydroxyurea for 20 hours.

Conjugation

unconjugated

Clone Number

PS01-17

RRID

AB_3072309

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IF-Cell

  • 1:200

Target

Function

Checkpoint kinase 1, commonly referred to as Chk1, is a serine/threonine-specific protein kinase that, in humans, is encoded by the CHEK1 gene. Chk1 coordinates the DNA damage response (DDR) and cell cycle checkpoint response. Activation of Chk1 results in the initiation of cell cycle checkpoints, cell cycle arrest, DNA repair and cell death to prevent damaged cells from progressing through the cell cycle. Checkpoint kinases (Chks) are protein kinases that are involved in cell cycle control. Two checkpoint kinase subtypes have been identified, Chk1 and Chk2. Chk1 is a central component of genome surveillance pathways and is a key regulator of the cell cycle and cell survival. Chk1 is required for the initiation of DNA damage checkpoints and has recently been shown to play a role in the normal (unperturbed) cell cycle. Chk1 impacts various stages of the cell cycle including the S phase, G2/M transition and M phase. In addition to mediating cell cycle checkpoints, Chk1 also contributes to DNA repair processes, gene transcription, egg production, embryo development, cellular responses to HIV infection and somatic cell viability.

Background References

1. Klomp JE et al. CHK1 protects oncogenic KRAS-expressing cells from DNA damage and is a target for pancreatic cancer treatment. Cell Rep. 2021 Nov

2. Wen Y et al. EZH2 activates CHK1 signaling to promote ovarian cancer chemoresistance by maintaining the properties of cancer stem cells. Theranostics. 2021 Jan

Subcellular Location

Nucleus, Chromosome, Cytoplasm, cytoskeleton, microtubule organizing center, centrosome.

UNIPROT

SwissProt: O14757 Human

SwissProt: O35280 Mouse

Synonyms

C85740 antibody

Cell cycle checkpoint kinase antibody

Checkpoint , S. pombe, homolog of, 1 antibody

Checkpoint kinase 1 antibody

Checkpoint kinase 1 homolog (S. pombe) antibody

CHEK 1 antibody

Chek1 antibody

Chk 1 antibody

Chk1 antibody

CHK1 checkpoint homolog (S. pombe) antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-Chk1 (S345) on different lysates with Rabbit anti-Phospho-Chk1 (S345) antibody (<a href="/products/HA721189" style="font-weight: bold;text-decoration: underline;">HA721189</a>) at 1/1,000 dilution.<br /><br />Lane 1: HEK-293 cell lysate<br />Lane 2: HEK-293 treated with 200nM Calyculin A for 1 hour cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 54 kDa<br />Observed band size: 56 kDa<br /><br />Exposure time: 59 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721189" style="font-weight: bold;text-decoration: underline;">HA721189</a>) at 1/1,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of Phospho-Chk1 (S345) on different lysates with Rabbit anti-Phospho-Chk1 (S345) antibody (HA721189) at 1/1,000 dilution.

    Lane 1: HEK-293 cell lysate
    Lane 2: HEK-293 treated with 200nM Calyculin A for 1 hour cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 54 kDa
    Observed band size: 56 kDa

    Exposure time: 59 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721189) at 1/1,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-Chk1 (S345) on different lysates with Rabbit anti-Phospho-Chk1 (S345) antibody (<a href="/products/HA721189" style="font-weight: bold;text-decoration: underline;">HA721189</a>) at 1/1,000 dilution.<br /><br />Lane 1: NIH/3T3 cell lysate<br />Lane 2: NIH/3T3 treated with UV for 20 minutes then recover for 2 hours cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 54 kDa<br />Observed band size: 56 kDa<br /><br />Exposure time: 1 minute; ECL: K1802;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721189" style="font-weight: bold;text-decoration: underline;">HA721189</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of Phospho-Chk1 (S345) on different lysates with Rabbit anti-Phospho-Chk1 (S345) antibody (HA721189) at 1/1,000 dilution.

    Lane 1: NIH/3T3 cell lysate
    Lane 2: NIH/3T3 treated with UV for 20 minutes then recover for 2 hours cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 54 kDa
    Observed band size: 56 kDa

    Exposure time: 1 minute; ECL: K1802;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721189) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-Chk1 (S345) on different lysates with Rabbit anti-Phospho-Chk1 (S345) antibody (<a href="/products/HA721189" style="font-weight: bold;text-decoration: underline;">HA721189</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HeLa treated with Etoposide cell lysate<br />Lane 3: HeLa cell lysate<br />Lane 4: HeLa treated with Hydroxyurea cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 54 kDa<br />Observed band size: 56 kDa<br /><br />Exposure time: 5 minutes;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721189" style="font-weight: bold;text-decoration: underline;">HA721189</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:300,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of Phospho-Chk1 (S345) on different lysates with Rabbit anti-Phospho-Chk1 (S345) antibody (HA721189) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HeLa treated with Etoposide cell lysate
    Lane 3: HeLa cell lysate
    Lane 4: HeLa treated with Hydroxyurea cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 54 kDa
    Observed band size: 56 kDa

    Exposure time: 5 minutes;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721189) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Immunocytochemistry analysis of HeLa cells treated with or without 4mM Hydroxyurea for 20 hours labeling Phospho-Chk1 (S345) with Rabbit anti-Phospho-Chk1 (S345) antibody (<a href="/products/HA721189" style="font-weight: bold;text-decoration: underline;">HA721189</a>) at 1/200 dilution. Image shown an increased nuclear staining upon Hydroxyurea treatment.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at 37 ℃, permeabilized with 0.1% Triton X-100 in PBS permeabilization for 5 minutes, and then blocked with 2% negative goat serum for 60 minutes at room temperature. Cells were then incubated with Rabbit anti-Phospho-Chk1 (S345) antibody (<a href="/products/HA721189" style="font-weight: bold;text-decoration: underline;">HA721189</a>) at 1/200 dilution in 1% BSA overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    ☑ Cell treatment (CT)

    Immunocytochemistry analysis of HeLa cells treated with or without 4mM Hydroxyurea for 20 hours labeling Phospho-Chk1 (S345) with Rabbit anti-Phospho-Chk1 (S345) antibody (HA721189) at 1/200 dilution. Image shown an increased nuclear staining upon Hydroxyurea treatment.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at 37 ℃, permeabilized with 0.1% Triton X-100 in PBS permeabilization for 5 minutes, and then blocked with 2% negative goat serum for 60 minutes at room temperature. Cells were then incubated with Rabbit anti-Phospho-Chk1 (S345) antibody (HA721189) at 1/200 dilution in 1% BSA overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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