製品の概要

  • 製品名Anti-ZW10 antibody
    ZW10 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to ZW10
  • アプリケーション適用あり: IHC-P, WB, ICC/IF, IPmore details
  • 種交差性
    交差種: Human
    交差が予測される動物種: Mouse, Rat, Dog, Xenopus laevis
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human ZW10.

    (Peptides available as ab23436, ab23437 and ab23436.)

  • ポジティブ・コントロール
    • Jurkat whole cell lysate

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab21582 in the following tested applications.

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

アプリケーション Abreviews 特記事項
IHC-P 1/150. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 88 kDa (predicted molecular weight: 88 kDa).
ICC/IF 1/100.
IP Use at an assay dependent concentration.

ターゲット情報

  • Information by UniProt
  • 参照データベース
  • 別名
    • Centromere/kinetochore protein zw10 antibody
    • Centromere/kinetochore protein zw10 homolog antibody
    • HZW 10 antibody
    • HZW10 antibody
    • Kinetochore associated homolog antibody
    • KNTC1AP antibody
    • MGC149821 antibody
    • Zeste white 10 antibody
    • Zeste white 10 homolog antibody
    • ZW 10 antibody
    • ZW 10 homolog centromere/kinetochore protein antibody
    • ZW 10 kinetochore associated homolog antibody
    • ZW10 (Drosophila) homolog centromere/kinetochore protein antibody
    • Zw10 antibody
    • ZW10 homolog centromere/kinetochore protein (Drosophila) antibody
    • ZW10 homolog centromere/kinetochore protein antibody
    • ZW10 kinetochore associated homolog antibody
    • ZW10, kinetochore associated, homolog (Drosophila) antibody
    • ZW10_HUMAN antibody
    see all

Anti-ZW10 antibody 画像

  • All lanes : Anti-ZW10 antibody (ab21582) at 1 µg/ml

    Lane 1 : Jurkat whole cell lysate (ab7899)
    Lane 2 : Jurkat whole cell lysate (ab7899) with ZW10 peptide (ab23436) at 1 µg/ml

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG(Alexa Fluor® 680) at 1/10,000 dilution

    Performed under reducing conditions.

    Predicted band size : 88 kDa
    Observed band size : 88 kDa
    Additional bands at : 45 kDa (possible cleavage fragment,cross reactivity),75 kDa (possible cleavage fragment,cross reactivity).
    This antibody detects a band at approximately 88kDa that corresponds in size to ZW10. It also detects bands at 75 and 45kDa that may be degradation products or crossreactivity. All of these bands are competed away by the addition of the immunizing peptide, suggesting that they are specific interactions.
  • ZW10 was immunoprecipitated using 0.5mg Jurkat whole cell extract, 5µg of Rabbit polyclonal to ZW10 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, Jurkat 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 ab21582.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 88kDa: ZW10
  • Panel one shows staining of HeLa cells by anti-ZW10 (ab21582) in green and DAPI in blue. The second panel shows staining by anti-ZW10 in green and SH-CREST in red, which stains the centromeres. In most cells at interphase, ZW10 is present diffusely in the cytoplasm. In prometaphase, it is associated with the kinetochore. Fix cells for 30 minutes on ice in 4% formaldehyde in PEM. Quench autofluorescence 2 x 5 minutes with 1 mg/ml Na borohydride or 100 mM ammonium chloride in PEM. Permeablize 30 minutes with 0.5% TX-100 in PEM. Block 30 minutes in 5% milk in TBST. Primary antibody incubated overnight at 4oC diluted 1/100 in 5% milk in TBST. Secondary antibody 1 hour at RT diluted in 5% milk in TBST. Post-fix 20 min. on ice in 4% formaldehyde in PEM. Quench autofluorescence 2 x 5 minutes with ammonium chloride in PEM. Counterstain with DAPI in TBST. Mount with ProLong Gold antifade reagent from Invitrogen. Notes: Ample washing between each step. TBST = Tris buffered saline +Fix cells for 30 minutes on ice in 4% formaldehyde in PEM. Quench autofluorescence 2 x 5 minutes with 1 mg/ml Na borohydride or 100 mM ammonium chloride in PEM. Permeablize 30 minutes with 0.5% TX-100 in PEM. Block 30 minutes in 5% milk in TBST. Primary antibody incubated overnight at 4oC diluted 1/100 in 5% milk in TBST. Secondary antibody 1 hour at RT diluted in 5% milk in TBST. Post-fix 20 min. on ice in 4% formaldehyde in PEM. Quench autofluorescence 2 x 5 minutes with ammonium chloride in PEM. Counterstain with DAPI in TBST. Mount with ProLong Gold antifade reagent from Invitrogen. Notes: Ample washing between each step. TBST = Tris buffered saline + 0.1% Tween. PEM = 80 mM K-PIPES, pH 6.8, 5 mM EGTA, 2 mM MgCl2.
  • Image courtesy of Human Protein Atlas ab215820 staining ZW10 in Human stomach, showing a strong staining pattern in gastric pits with lower levels in the basement membrane. Paraffin embedded human stomach tissue was incubated with ab21582 (1/150 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.

    ab21582 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org

  • ab21582 (1/200) staining ZW10 in asynchronous mouse 10T1/2 cells (green). Cells were pre-extracted with PEM (1min) at room temperature, fixed in paraformaldehyde (10 mins), washed in KB+ buffer and counterstained with DAPI in order to highlight the DNA/nucleus (red). Please note that ab21582 might only perform well when using this specific protocol. Please refer to abreview for further experimental details.

    See Abreview

Anti-ZW10 antibody (ab21582) 使用論文

This product has been referenced in:
  • Zhang G  et al. Distinct domains in Bub1 localize RZZ and BubR1 to kinetochores to regulate the checkpoint. Nat Commun 6:7162 (2015). ICC/IF ; Human . Read more (PubMed: 26031201) »
  • Maldonado M & Kapoor TM Constitutive Mad1 targeting to kinetochores uncouples checkpoint signalling from chromosome biorientation. Nat Cell Biol 13:475-82 (2011). ICC/IF ; Human . Read more (PubMed: 21394085) »

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 Human Tissue sections (HCV infected patients liver biopsy)
Specification HCV infected patients liver biopsy
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated
Permeabilization No
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Dr. Saira Khalid

Verified customer

投稿 Dec 13 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (GM08505 cell line)
Specification GM08505 cell line
Fixative Paraformaldehyde
Permeabilization Yes - Triton
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 37°C
Username

Mr. Faisal Alzahrani

Verified customer

投稿 Oct 18 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Hela)
Specification Hela
Fixative Formaldehyde
Blocking step Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
Username

Abcam user community

Verified customer

投稿 Aug 01 2012

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (10T1/2)
Specification 10T1/2
Fixative PEM then Paraformaldehyde
Permeabilization Yes - 0.5% Triton X100 in PBS
Username

Dr. Kirk McManus

Verified customer

投稿 May 18 2009

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Methanol
Permeabilization No
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Dec 11 2008

Application Western blot
Sample Human Cell lysate - whole cell (HeLa)
Loading amount 25 µg
Specification HeLa
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Sep 02 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa CCL2)
Specification HeLa CCL2
Fixative Paraformaldehyde
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Dr. Kyung Lee

Verified customer

投稿 Jan 04 2007

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Username

Dr. Kirk McManus

Verified customer

投稿 Dec 18 2006

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