Anti-Histone H2A (phospho S1) + Histone H4 (phospho S1)抗体(ab14723)

概述

  • 产品名称
    Anti-Histone H2A (phospho S1) + Histone H4 (phospho S1)抗体
  • 描述
    兔多克隆抗体to Histone H2A (phospho S1) + Histone H4 (phospho S1)
  • 特异性
    By peptide array ab14723 has been shown to detect both Histone H2A (phospho S1) and Histone H4 (phospho S1) peptides and not the equivalent unmodified peptides (please see peptide array image on this datasheet). However, by western blot, this antibody shows a preference for Histone H4 (phospho S1) as seen in the western blot images on this datasheet. If you have further questions about the specificity of ab14723, please contact our technical support team, who will be happy to help.
  • 经测试应用
    适用于: WB, IP, ICC/IF, IHC-P, PepArrmore details
  • 种属反应性
    与反应: Human, Saccharomyces cerevisiae
  • 免疫原

    Synthetic peptide corresponding to Histone H4 aa 1-100 (phospho S1) conjugated to keyhole limpet haemocyanin. Also UniProt ID: P04908, Q93077, Q99878 (Histone H2A)
    Database link: P62805
    (Peptide available as ab14724)

性能

  • 形式
    Liquid
  • 存放说明
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • 存储溶液
    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 1% BSA
  • Concentration information loading...
  • 纯度
    Immunogen affinity purified
  • 纯化说明
    This antibody was affinity purified using the phospho peptide ab14724, it was then cross-affinity purified using the non-phospho peptide ab14725 in order to remove reactivity with the non-phosphorylated epitope.
  • 克隆
    多克隆
  • 同种型
    IgG
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab14723 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB 1/1000. Detects a band of approximately 15 kDa (predicted molecular weight: 11 kDa).

This antibody also gave a positive result in ELISA against the immunising peptide (ab14724). It gave a negative result in ELISA against the non-modified equivalent peptide (ab14725).

IP 1/50. See Abreview.
ICC/IF Use at an assay dependent concentration.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
PepArr Use a concentration of 0.1 - 0.01 µg/ml.

靶标

图片

  • Peptide Array analysis of ab14723 at 1:5000 dilution against 40.0 – 0.6 µM peptides: Histone H2A unmodified peptide (Lane 1), Histone H2A (phospho S1) peptide (Lane 2), Histone H4 unmodified peptide (Lane 3) and Histone H4 (phospho S1) peptide (Lane 4). Results show strong binding to the phosphorylated, but not the unmodified versions of these peptides. 

  • All lanes : Anti-Histone H2A (phospho S1) + Histone H4 (phospho S1) antibody (ab14723) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate at 10 µg
    Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate - Colcemid Treated at 2.5 µg
    Lane 4 : Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg
    Lane 5 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg

    Secondary
    Peroxidase AffiniPure Goat Anti-Rabbit IgG (H+L) (ab201489) at 1/50000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 11 kDa
    Observed band size : 14 kDa (why is the actual band size different from the predicted?)


    Exposure time : 90 seconds

    This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab14723 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

  • All lanes : Anti-Histone H2A (phospho S1) + Histone H4 (phospho S1) antibody (ab14723) at 1/1000 dilution

    Lane 1 : YDH8 cka 2-8 ts grown at 30 degrees C
    Lane 2 : YDH8 cka 2-8 ts grown at 38 degrees C

    Secondary
    HRP conjugated anti-Rabbit

    Predicted band size : 11 kDa
    Observed band size : 15 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 48 kDa. We are unsure as to the identity of these extra bands.

    This image was submitted as part of a review by Meredith Calvert

    Cell lysates were prepared from a Casein Kinase 2 temperature sensitive Saccharomyces cerevisiae strain (YDH8 cka2-8ts) grown at either 30°C or 38°C. The kinase is active at °C and inactive at 38°C. Lysates were prepared from asynchronous log-phase cultures and 300µl run on a gel. The Western blot was blocked in milk for 20  minutes and incubated with a 1/1000 dilution of ab14723 for 16 hours. ab14723 detected a band of 15kDa representing Histone H4 (phospho S1) in the lysates grown at 30°C. 

     

  • IHC image of Histone H2A (phospho S1) + Histone H4 (phospho S1) staining in human breast carcinoma FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab14723, 1µg/ml, for 8 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

文献

This product has been referenced in:
  • Zhang ZH  et al. Dynamics of histone H2A, H4 and HS1ph during spermatogenesis with a focus on chromatin condensation and maturity of spermatozoa. Sci Rep 6:25089 (2016). WB . Read more (PubMed: 27121047) »
  • Tjeertes JV  et al. Screen for DNA-damage-responsive histone modifications identifies H3K9Ac and H3K56Ac in human cells. EMBO J 28:1878-89 (2009). WB ; Human . Read more (PubMed: 19407812) »

See all 2 Publications for this product

客户评价及客户问答

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ChIP
Sample
Saccharomyces cerevisiae Cell lysate - nuclear (S288c)
Specification
S288c
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 15 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% Fromaldehyde
Detection step
Real-time PCR
Username

Mr. Harsh Ja

Verified customer

提交于 Aug 02 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Saccharomyces cerevisiae Cell lysate - whole cell (S288c)
Loading amount
25 µg
Specification
S288c
Gel Running Conditions
Reduced Denaturing (15% gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
Username

Mr. Harsh Ja

Verified customer

提交于 Aug 02 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Saccharomyces cerevisiae Cell lysate - nuclear (BY4147 background)
Total protein in input
15 µg
Specification
BY4147 background
Treatment
1% MMS (DNA damaging agent)
Immuno-precipitation step
Protein A/G
Username

Abcam user community

Verified customer

提交于 Jul 26 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Saccharomyces cerevisiae Cell (BY4147 background)
Specification
BY4147 background
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.25% Triton X-100 in PBS
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Jun 01 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ChIP on chip
Sample
Saccharomyces cerevisiae Cell lysate - nuclear (W303 background)
Specification
W303 background
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 20 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% Formaldehyde
Detection step
Microarray
Username

Abcam user community

Verified customer

提交于 May 31 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ChIP
Sample
Saccharomyces cerevisiae Cell lysate - nuclear (Yeast chromatin lysate)
Specification
Yeast chromatin lysate
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 20 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% Formaldehyde
Detection step
Semiquantitative PCR
Username

Abcam user community

Verified customer

提交于 May 31 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Saccharomyces cerevisiae Cell lysate - nuclear (Yeast whole cell extract)
Loading amount
10 µg
Specification
Yeast whole cell extract
Gel Running Conditions
Reduced Denaturing (15% gel)
Blocking step
Milk as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
Username

Abcam user community

Verified customer

提交于 May 26 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Saccharomyces cerevisiae Cell lysate - whole cell (Asynchronous log-phase culture)
Specification
Asynchronous log-phase culture
Blocking step
Milk as blocking agent for 20 minute(s) · Concentration: 5%
Username

Dr. Meredith Calvert

Verified customer

提交于 Feb 23 2006

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

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