Anti-Separase抗体[XJ11-1B12] (ab16170)

概述

  • 产品名称
    Anti-Separase抗体[XJ11-1B12]
    参阅全部 Separase 一抗
  • 描述
    小鼠单克隆抗体[XJ11-1B12] to Separase
  • 经测试应用
    适用于: Flow Cyt, WB, ICC/IFmore details
  • 种属反应性
    与反应: Human
  • 免疫原

    Maltose binding protein fusion protein, corresponding to C terminal amino acids 1866-1996 of Human Separase.

  • 阳性对照
    • This antibody gave a positive signal in MDA-MB-231 Whole Cell Lysate.
  • 常规说明

    This antibody clone is manufactured by Abcam.


    Separase antibodies can be used as a specific marker for centrosomes of mitotic cells. The staining of separase in centrosomes can be detected from prophase of mitosis up until anaphase.

性能

  • 形式
    Liquid
  • 存放说明
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • 存储溶液
    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • 纯度
    IgG fraction
  • Primary antibody说明
    Separase antibodies can be used as a specific marker for centrosomes of mitotic cells. The staining of separase in centrosomes can be detected from prophase of mitosis up until anaphase.
  • 克隆
    单克隆
  • 克隆编号
    XJ11-1B12
  • 同种型
    IgG1
  • 轻链类型
    kappa
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab16170 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
Flow Cyt Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
WB Use a concentration of 10 µg/ml. Detects a band of approximately 170 kDa (predicted molecular weight: 233 kDa).
ICC/IF Use at an assay dependent concentration. Used at a dilution of 1/100 for 1 hr on HeLa cells (see Abreview).

靶标

  • 功能
    Caspase-like protease, which plays a central role in the chromosome segregation by cleaving the SCC1/RAD21 subunit of the cohesin complex at the onset of anaphase. During most of the cell cycle, it is inactivated by different mechanisms.
  • 序列相似性
    Belongs to the peptidase C50 family.
  • 翻译后修饰
    Autocleaves. This function, which is not essential for its protease activity, is unknown.
    Phosphorylated by CDK1. There are 8 Ser/Thr phosphorylation sites. Among them, Ser-1126 phosphorylation is the major site, which conducts to the enzyme inactivation.
  • 细胞定位
    Cytoplasm. Nucleus.
  • Information by UniProt
  • 数据库链接
  • 别名
    • Caspase like protein ESPL1 antibody
    • Caspase-like protein ESPL1 antibody
    • ESP 1 antibody
    • ESP1 antibody
    • ESPL 1 antibody
    • Espl1 antibody
    • ESPL1_HUMAN antibody
    • Extra spindle poles like 1 antibody
    • Extra spindle poles like 1 protein antibody
    • Extra spindle poles-like 1 protein antibody
    • FLJ46492 antibody
    • KIAA0165 antibody
    • Separase antibody
    • Separin antibody
    • Similar to fission yeast cut1and gene antibody
    • SSE antibody
    see all

图片

  • ab16170 stained MCF7 cells. The cells were 100% methanol fixed for 5 minutes at -20°C and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab16170 at 10µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  • Overlay histogram showing HeLa cells stained with ab16170 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab16170, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • Anti-Separase antibody [XJ11-1B12] (ab16170) at 10 µg/ml + MDA-MB-231 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 20 µg

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 233 kDa
    Observed band size : 170 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 100 kDa,70 kDa (possible cleavage fragment). We are unsure as to the identity of these extra bands.

    Exposure time : 3 minutes

文献

This product has been referenced in:
  • Seo MY  et al. Integrity of the Pericentriolar Material Is Essential for Maintaining Centriole Association during M Phase. PLoS One 10:e0138905 (2015). WB . Read more (PubMed: 26407333) »
  • Kim J  et al. PLK1 regulation of PCNT cleavage ensures fidelity of centriole separation during mitotic exit. Nat Commun 6:10076 (2015). WB . Read more (PubMed: 26647647) »

See all 9 Publications for this product

客户评价及客户问答

Thank you for calling us and for alerting us to the problem you are experiencing with our product. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly. If you wouldn't mind, could yo...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Hela)
Loading amount
12500 cells
Specification
Hela
Treatment
Transfected with siRNA duplexes (control or targetting separase), subsequent nocodazole arrest
Gel Running Conditions
Reduced Denaturing (4-12% PAGE)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

提交于 Sep 16 2010

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (MDA-MB-231 breast cancer cell line)
Total protein in input
40 µg
Specification
MDA-MB-231 breast cancer cell line
Immuno-precipitation step
Protein G
Username

Abcam user community

Verified customer

提交于 Oct 10 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (MDA-MB-231 breast cancer cell line)
Loading amount
40 µg
Specification
MDA-MB-231 breast cancer cell line
Gel Running Conditions
Reduced Denaturing (gel 8 %)
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

提交于 Oct 10 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Methanol
Blocking step
Serum as blocking agent for 45 minute(s) · Concentration: 5%
Username

Dr. Ashraf Dallol

Verified customer

提交于 Jun 22 2006

Thank you for your enquiry. Here we are sending a general Western blot protocol. The information is only intended as a guide. The researchers should determine what protocol best meets their needs: Sample Preparation 1. Remove media and wash ...

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

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