Anti-HP1 gamma (phospho S93)抗体(ab45270)

概述

  • 产品名称
    Anti-HP1 gamma (phospho S93)抗体
    参阅全部 HP1 gamma 一抗
  • 描述
    兔多克隆抗体to HP1 gamma (phospho S93)
  • 经测试应用
    适用于: IHC-Fr, ICC/IF, IP, WB, IHC-Pmore details
  • 种属反应性
    与反应: Human, Rhesus monkey
    预测可用于: Mouse, Cow, Orangutan
  • 免疫原

    Synthetic peptide corresponding to Human HP1 gamma aa 50-150 conjugated to Keyhole Limpet Haemocyanin (KLH).
    Database link: Q13185
    (Peptide available as ab45386, ab45386, ab45386)

  • 阳性对照
    • ab45270 gave a positive result in the following Human Whole Cell Lysates: Hela, Jurkat, A431, HEK293, HepG2, MCF-7 and SHSY-5Y.

性能

应用

Our Abpromise guarantee covers the use of ab45270 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
IHC-Fr Use at an assay dependent concentration. PubMed: 25216677
ICC/IF Use a concentration of 5 µg/ml.
IP Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 24 kDa (predicted molecular weight: 21 kDa).
IHC-P Use a concentration of 1 µg/ml.

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 (phospho S93) antibody 图像

  • ICC/IF image of ab45270 stained human HeLa cells. The cells were methanol fixed (5 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab45270, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

  • IHC image of HP1 gamma (phospho S83) staining in Human lung adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica 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 ab45270, 1µg/ml, for 15 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.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • All lanes : Anti-HP1 gamma (phospho S93) antibody (ab45270) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 4 : HEK293 Human embryonic kidney cell line Whole Cell Lysate
    Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 6 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 7 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
    Lane 8 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with Human HP1 gamma peptide (ab51840) at 1 µg/ml
    Lane 9 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate with Human HP1 gamma peptide (ab51840) at 1 µg/ml
    Lane 10 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate with Human HP1 gamma peptide (ab51840) at 1 µg/ml
    Lane 11 : HEK293 Human embryonic kidney cell line Whole Cell Lysate with Human HP1 gamma peptide (ab51840) at 1 µg/ml
    Lane 12 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate with Human HP1 gamma peptide (ab51840) at 1 µg/ml
    Lane 13 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate (ab28419) with Human HP1 gamma peptide (ab51840) at 1 µg/ml
    Lane 14 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate (ab45968) with Human HP1 gamma peptide (ab51840) at 1 µg/ml
    Lane 15 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with Human HP1 gamma (phospho S93) peptide (ab45386) at 1 µg/ml
    Lane 16 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate with Human HP1 gamma (phospho S93) peptide (ab45386) at 1 µg/ml
    Lane 17 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate with Human HP1 gamma (phospho S93) peptide (ab45386) at 1 µg/ml
    Lane 18 : HEK293 Human embryonic kidney cell line Whole Cell Lysate with Human HP1 gamma (phospho S93) peptide (ab45386) at 1 µg/ml
    Lane 19 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate with Human HP1 gamma (phospho S93) peptide (ab45386) at 1 µg/ml
    Lane 20 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate with Human HP1 gamma (phospho S93) peptide (ab45386) at 1 µg/ml
    Lane 21 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate with Human HP1 gamma (phospho S93) peptide (ab45386) at 1 µg/ml

    Lysates/proteins at 10 µg per lane.

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 21 kDa
  • HP1 gamma was immunoprecipitated using 0.5mg HepG2 whole cell extract, 5µg of Rabbit polyclonal to HP1 gamma and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, HepG2 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab45270.
    Secondary: Clean-Blot IP Detection Reagent (HRP) at 1/500 dilution.
    Band: 22kDa; HP1 gamma

Anti-HP1 gamma (phospho S93) antibody (ab45270)参考文献

This product has been referenced in:
  • Lastwika KJ  et al. Control of PD-L1 Expression by Oncogenic Activation of the AKT-mTOR Pathway in Non-Small Cell Lung Cancer. Cancer Res 76:227-38 (2016). Read more (PubMed: 26637667) »
  • Rielland M  et al. Senescence-associated SIN3B promotes inflammation and pancreatic cancer progression. J Clin Invest 124:2125-35 (2014). IHC-P ; Mouse . Read more (PubMed: 24691445) »

See all 4 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rhesus monkey Tissue sections (Colon)
Specification
Colon
Fixative
Z-FIX
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer
Permeabilization
Yes - 0.1% Triton-X-100
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
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提交于 Apr 11 2013

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Raw 264.7)
Loading amount
25 µg
Specification
Raw 264.7
Treatment
LPS 2 hours
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 23°C
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Verified customer

提交于 Aug 07 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Recombinant protein (Unmodified recombinant protein)
Loading amount
0.2 µg
Specification
Unmodified recombinant protein
Treatment
incubated with kinase and 200uM ATP
Gel Running Conditions
Reduced Denaturing (10)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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Verified customer

提交于 Jun 08 2012

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

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