概述

  • 产品名称Anti-CRM1抗体
    参阅全部 CRM1 一抗
  • 描述
    兔多克隆抗体to CRM1
  • 经测试应用适用于: IHC-P, ICC/IF, WBmore details
  • 种属反应性
    与反应: Human
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 1000 to the C-terminus of Human CRM1.

    (Peptide available as ab25749.)

  • 阳性对照
    • ab24189 gave a positive result in HeLa whole cell lysate. This antibody also gave a positive signal in IHC in human hippocampus tissue sections.

性能

应用

Our Abpromise guarantee covers the use of ab24189 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-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 100 kDa (predicted molecular weight: 123 kDa).

Abcam recommends using 3% milk as the blocking agent.

靶标

  • 功能Mediates the nuclear export of cellular proteins (cargos) bearing a leucine-rich nuclear export signal (NES) and of RNAs. In the nucleus, in association with RANBP3, binds cooperatively to the NES on its target protein and to the GTPase RAN in its active GTP-bound form (Ran-GTP). Docking of this complex to the nuclear pore complex (NPC) is mediated through binding to nucleoporins. Upon transit of an nuclear export complex into the cytoplasm, disassembling of the complex and hydrolysis of Ran-GTP to Ran-GDP (induced by RANBP1 and RANGAP1, respectively) cause release of the cargo from the export receptor. The directionality of nuclear export is thought to be conferred by an asymmetric distribution of the GTP- and GDP-bound forms of Ran between the cytoplasm and nucleus. Involved in U3 snoRNA transport from Cajal bodies to nucleoli. Binds to late precursor U3 snoRNA bearing a TMG cap. Several viruses, among them HIV-1, HTLV-1 and influenza A use it to export their unspliced or incompletely spliced RNAs out of the nucleus. Interacts with, and mediates the nuclear export of HIV-1 Rev and HTLV-1 Rex proteins. Involved in HTLV-1 Rex multimerization.
  • 组织特异性Expressed in heart, brain, placenta, lung, liver, skeletal muscle, pancreas, spleen, thymus, prostate, testis, ovary, small intestine, colon and peripheral blood leukocytes. Not expressed in the kidney.
  • 序列相似性Belongs to the exportin family.
    Contains 10 HEAT repeats.
    Contains 1 importin N-terminal domain.
  • 翻译后修饰Phosphorylated upon DNA damage, probably by ATM or ATR.
  • 细胞定位Cytoplasm. Nucleus > nucleoplasm. Nucleus > Cajal body. Nucleus > nucleolus. Located in the nucleoplasm, Cajal bodies and nucleoli. Shuttles between the nucleus/nucleolus and the cytoplasm.
  • Information by UniProt
  • 数据库链接
  • 别名
    • Chromosome region maintenance 1 protein homolog antibody
    • CRM 1 antibody
    • CRM1 homolog antibody
    • DKFZp686B1823 antibody
    • emb antibody
    • Exp 1 antibody
    • Exp1 antibody
    • Exportin 1 antibody
    • Exportin-1 antibody
    • Exportin1 antibody
    • XPO 1 antibody
    • xpo1 antibody
    • XPO1_HUMAN antibody
    see all

Anti-CRM1 antibody 图像

  • Anti-CRM1 antibody (ab24189) at 1 µg/ml + HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252) at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 123 kDa
    Observed band size : 120 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 73 kDa (possible non-specific binding).

    Exposure time : 90 seconds

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab24189 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

  • ICC/IF image of ab24189 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab24189 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • IHC image of CRM1 staining in Human normal hippocampus formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ 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 ab24189, 5µg/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-CRM1 antibody (ab24189) at 1 µg/ml

    Lane 1 : HeLa whole cell lysate
    Lane 2 : HeLa nuclear lysate
    Lane 3 : HeLa whole cell lysate with Human CRM1 peptide (ab25749) at 1 µg/ml
    Lane 4 : HeLa nuclear cell lysate with Human CRM1 peptide (ab25749) at 1 µg/ml

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 123 kDa
    Observed band size : 100 kDa (why is the actual band size different from the predicted?)

    This image is courtesy of an anonymous Abreview

    ab24189 detects a band of ~ 100 kDa in HeLa whole and HeLa nuclear cell lysates. This is somewhat smaller than the predicted band size according to Swissprot (123 kDa), however as both these bands are specifically blocked by the addition of the immunizing peptide (ab25749) we believe they represent CRM1.

    See Abreview

Anti-CRM1 antibody (ab24189)参考文献

This product has been referenced in:
  • Fleta-Soriano E  et al. The myxobacterial metabolite ratjadone A inhibits HIV infection by blocking the Rev/CRM1-mediated nuclear export pathway. Microb Cell Fact 13:17 (2014). WB ; Human . Read more (PubMed: 24475978) »
  • Bacot-Davis VR  et al. Solution structures of Mengovirus Leader protein, its phosphorylated derivatives, and in complex with nuclear transport regulatory protein, RanGTPase. Proc Natl Acad Sci U S A 111:15792-7 (2014). Read more (PubMed: 25331866) »

See all 4 Publications for this product

Product Wall

I would like to reassure you that our antibodies have all been tested successfully and would be covered by our guarantee in the applications listed on the datasheets. Therefore, any anti-CRM1 antibody tested in the species and application you are using...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa (cervical cancer cell line))
Specification HeLa (cervical cancer cell line)
Fixative Paraformaldehyde/methanol
Permeabilization Yes - Tris buffered saline with 0.5% triton X100
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Sep 01 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HeLa cells)
Loading amount 10 µg
Specification HeLa cells
Gel Running Conditions Reduced Denaturing (4-20% gradient gel)
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 Sep 01 2010

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