使用敲除细胞株进行验证

Anti-HP1 gamma抗体- ChIP Grade (ab10480)

概述

  • 产品名称Anti-HP1 gamma抗体- ChIP Grade
    参阅全部 HP1 gamma 一抗
  • 描述
    兔多克隆抗体to HP1 gamma - ChIP Grade
  • 经测试应用适用于: WBmore details
  • 种属反应性
    与反应: Mouse, Human
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Mouse HP1 gamma.

    .

  • 阳性对照
    • This antibody gave a positive signal in the following whole cell lysates: HeLa; MCF7; HEK293.

性能

应用

Our Abpromise guarantee covers the use of ab10480 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 1/500 - 1/1000. Detects a band of approximately 24 kDa (predicted molecular weight: 21 kDa).

靶标

  • 功能Seems to be involved in transcriptional silencing in heterochromatin-like complexes. Recognizes and binds histone H3 tails methylated at 'Lys-9', leading to epigenetic repression. May contribute to the association of the heterochromatin with the inner nuclear membrane through its interaction with lamin B receptor (LBR). Involved in the formation of functional kinetochore through interaction with MIS12 complex proteins.
  • 序列相似性Contains 2 chromo domains.
  • 翻译后修饰Phosphorylated by PIM1. Phosphorylated during interphase and possibly hyper-phosphorylated during mitosis.
  • 细胞定位Nucleus. Associates with euchromatin and is largely excluded from constitutive heterochromatin. May be associated with microtubules and mitotic poles during mitosis.
  • Information by UniProt
  • 数据库链接
  • 别名
    • CBX 3 antibody
    • CBX3 antibody
    • CBX3_HUMAN antibody
    • Chromobox homolog 3 (HP1 gamma homolog, Drosophila) antibody
    • Chromobox homolog 3 antibody
    • Chromobox protein homolog 3 antibody
    • GAMMA antibody
    • HECH antibody
    • Heterochromatin like protein 1 antibody
    • Heterochromatin protein 1 homolog gamma antibody
    • Heterochromatin protein HP1 gamma antibody
    • HP1 gamma antibody
    • HP1 gamma homolog antibody
    • HP1Hs gamma antibody
    • Modifier 2 protein antibody
    see all

Anti-HP1 gamma antibody - ChIP Grade 图像



  • Predicted band size : 21 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: HP1 gamma knockout HAP1 cell lysate (20 µg)

    Lane 3: NIH/3T3 cell lysate (20 µg)
    Lane 4: HeLa cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab10480 observed at 24 kDa. Red - loading control, ab8245, observed at 37 kDa.
    ab10480 was shown to recognize HP1 gamma when HP1 gamma knockout samples were used, along with additional cross-reactive bands. Wild-type and HP1 gamma knockout samples were subjected to SDS-PAGE. ab10480 and ab8245 (loading control to GAPDH) were both diluted 1/1000 and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • All lanes : Anti-HP1 gamma antibody - ChIP Grade (ab10480) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 21 kDa
    Observed band size : 24 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 49 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 12 minutes

  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 21 kDa
    Ab10480 recognizes the tagged recombinant HP1 gamma protein (ab88011) which has an expected molecular weight of ~50 kDa.

  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 21 kDa


    Exposure time : 30 seconds
    Ab10480 recognizes the tagged recombinant HP1 gamma protein (ab88011) which has an expected molecular weight of ~50 kDa.

Anti-HP1 gamma antibody - ChIP Grade (ab10480)参考文献

This product has been referenced in:
  • Blanc S  et al. Subcellular localization of ENS-1/ERNI in chick embryonic stem cells. PLoS One 9:e92039 (2014). WB ; Chicken . Read more (PubMed: 24643087) »
  • Kao TH  et al. Ectopic DNMT3L triggers assembly of a repressive complex for retroviral silencing in somatic cells. J Virol 88:10680-95 (2014). Read more (PubMed: 24991018) »

See all 7 Publications for this product

Product Wall

Application Western blot
Loading amount 15 µg
Gel Running Conditions Reduced Denaturing (7.5%)
Sample Human Cell lysate - nuclear (293T)
Specification 293T
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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提交于 Dec 16 2013

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris pH 9
Sample Mouse Tissue sections (Mouse 14.5dpc)
Specification Mouse 14.5dpc
Permeabilization No
Fixative Formaldehyde
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提交于 Nov 15 2013

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I am attaching few questions here, please answers these as these will help us to investigate what went wrong with these antibodies;

Sample preparation (X-ChIP/N-ChIP, cross-linking conditions, DNA fragment...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Skin, epidermis)
Specification Skin, epidermis
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate buffer pH 6.0
Permeabilization Yes - TBST
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 22°C
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提交于 Feb 16 2010

Application Immunocytochemistry
Sample Human Cultured Cells (Colorectal Cancer Cells)
Specification Colorectal Cancer Cells
Fixative Formaldehyde
Permeabilization Yes - Triton X-100
Blocking step Serum as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
Username

Abcam user community

Verified customer

提交于 Jan 06 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Colorectal Cancer Cells)
Loading amount 200 µg
Specification Colorectal Cancer Cells
Gel Running Conditions Reduced Denaturing (12% Gel)
Blocking step Milk as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 27°C
Username

Abcam user community

Verified customer

提交于 Jan 04 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Human Colon Cancer)
Specification Human Colon Cancer
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: DAKO Target Retrieval Solution
Permeabilization No
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 27°C
Username

Abcam user community

Verified customer

提交于 Nov 03 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HMEC human mammary epithelial cells)
Specification HMEC human mammary epithelial cells
Fixative Paraformaldehyde
Permeabilization Yes - fixation was combined with permeabilazation with 4%PF in PBS 0,1% Triton X-100
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
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Dr. Silvia Sirchia

Verified customer

提交于 Jun 09 2008

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