Anti-NTF97/Importin beta抗体[3E9] (ab2811)

概述

  • 产品名称Anti-NTF97/Importin beta抗体[3E9]
    参阅全部 NTF97/Importin beta 一抗
  • 描述
    小鼠单克隆抗体[3E9] to NTF97/Importin beta
  • 经测试应用适用于: Flow Cyt, ChIP, IP, Inhibition Assay, WB, ICC/IFmore details
  • 种属反应性
    与反应: Mouse, Rat, Cow, Dog, Human, Pig, African Green Monkey, Syrian Hamster
  • 免疫原

    Full length native protein (purified) corresponding to Cow NTF97/Importin beta. Purified from Bovine erythrocytes.

  • 阳性对照
    • WB: MDBK cell lysate ICC: MDBK cells, PTK cells

性能

  • 形式Liquid
  • 存放说明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • 存储溶液Preservative: 0.05% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • 纯度Protein A purified
  • Primary antibody说明The accumulation of proteins in the nucleus is mediated by short sequences of basic amino acids called nuclear localization sequences (NLSs). These sequences are necessary and sufficient to direct a protein or inert carrier to the nuclear interior. Nuclear protein import is accomplished by two sequential, energy dependent events. The first step, docking at the nuclear pore complex, requires a 54/56 kDa nuclear localization signal receptor (a-karyopherin, importin-a, SRP-a) and the nuclear transport factor p97 (NTF 97, b-karyopherin, importin-b). The second step, translocation across the nuclear envelope (NE), requires the GTP-binding protein, Ran/TC4.
  • 克隆单克隆
  • 克隆编号3E9
  • 同种型IgG2a
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab2811 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 1/100.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

ChIP Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
Inhibition Assay Use at an assay dependent concentration.
WB 1/5000. Detects a band of approximately 97 kDa (predicted molecular weight: 97 kDa).
ICC/IF 1/1000.

靶标

  • 功能Functions in nuclear protein import, either in association with an adapter protein, like an importin-alpha subunit, which binds to nuclear localization signals (NLS) in cargo substrates, or by acting as autonomous nuclear transport receptor. Acting autonomously, serves itself as NLS receptor. Docking of the importin/substrate complex to the nuclear pore complex (NPC) is mediated by KPNB1 through binding to nucleoporin FxFG repeats and the complex is subsequently translocated through the pore by an energy requiring, Ran-dependent mechanism. At the nucleoplasmic side of the NPC, Ran binds to importin-beta and the three components separate and importin-alpha and -beta are re-exported from the nucleus to the cytoplasm where GTP hydrolysis releases Ran from importin. The directionality of nuclear import is thought to be conferred by an asymmetric distribution of the GTP- and GDP-bound forms of Ran between the cytoplasm and nucleus. Mediates autonomously the nuclear import of ribosomal proteins RPL23A, RPS7 and RPL5. Binds to a beta-like import receptor binding (BIB) domain of RPL23A. In association with IPO7 mediates the nuclear import of H1 histone. In vitro, mediates nuclear import of H2A, H2B, H3 and H4 histones. In case of HIV-1 infection, binds and mediates the nuclear import of HIV-1 Rev. Imports PRKCI into the nucleus.
  • 序列相似性Belongs to the importin beta family.
    Contains 8 HEAT repeats.
    Contains 1 importin N-terminal domain.
  • 细胞定位Cytoplasm. Nucleus envelope.
  • Information by UniProt
  • 数据库链接
  • 别名
    • IMB 1 antibody
    • IMB1 antibody
    • IMB1_HUMAN antibody
    • Impnb antibody
    • Importin 90 antibody
    • Importin beta 1 antibody
    • Importin beta 1 subunit antibody
    • Importin subunit beta-1 antibody
    • Importin-90 antibody
    • IPOB antibody
    • Karyopherin beta 1 antibody
    • Karyopherin beta 1 subunit antibody
    • Karyopherin subunit beta-1 antibody
    • KPNB 1 antibody
    • Kpnb1 antibody
    • MGC2155 antibody
    • MGC2156 antibody
    • MGC2157 antibody
    • NTF 97 antibody
    • NTF97 antibody
    • Nuclear factor P97 antibody
    • Pore targeting complex 97 kDa subunit antibody
    • PTAC97 antibody
    see all

Anti-NTF97/Importin beta antibody [3E9] 图像

  • Flow cytometry analysis of NTF97/Importin beta in Jurkat cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/ml, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab2811 (1ug/test) for 40 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated secondary antibody and re-suspended in PBS for FACS analysis.

  • Flow cytometry analysis of NTF97/Importin beta in 3T3 cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/ml, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab2811 (1ug/test) for 40 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated secondary antibody and re-suspended in PBS for FACS analysis.

  • Flow cytometry analysis of NTF97/Importin beta in PC12 cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/ml, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab2811 (1ug/test) for 40 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated secondary antibody and re-suspended in PBS for FACS analysis.

  • All lanes : Anti-NTF97/Importin beta antibody [3E9] (ab2811) at 1/2000 dilution

    Lane 1 : Mouse lung whole tissue lysate
    Lane 2 : Mouse kidney whole tissue lysate
    Lane 3 : Mouse spleen whole tissue lysate

    Secondary
    HRP-conjugated Goat anti-mouse IgG polyclonal at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 97 kDa
    Observed band size : 97 kDa


    Exposure time : 3 minutes

    This image is courtesy of an anonymous Abreview

    See Abreview

  • All lanes : Anti-NTF97/Importin beta antibody [3E9] (ab2811) at 50

    Lane 1 : U251 cell lysate
    Lane 2 : HepG2 cell lysate
    Lane 3 : C6 cell lysate

    Lysates/proteins at 25 µg per lane.


    Predicted band size : 97 kDa
    Observed band size : 100 kDa (why is the actual band size different from the predicted?)
  • Immunolocalization of NTF97 in PTK cells using ab2811.
  • NIH3T3 cells were incubated for 4 minutes in PHEM/1%triton, washed for 2 minutes in 1x PHEM and fixed for 10 minutes at room temperature in 3.7% PFA containing 30mM sucrose. Following washing in PBS, the cells were incubated for 2 minutes in 100% Methanol at -20°C, then washed 3 times in PBS. The cells were then incubated with ab2811 (1/200) for 1 hour at room temperature. The image panel shows the nuclei stained with DAPI (blue) and the nuclear envalope and cytoplasm stained with ab2811 (green). 100x magnification.

  • Immunoprecipitation of Importin beta, in HeLa cells, using ab2811. Coomassie-stained 8% SDS-page gel was loaded with IP fractions obtained by incubating 2 mg of pre-cleared HeLa whole cell extracts with 4µg ab2811 or 4µg IgG (control). The Importin band (see arrow) was cut out of the gel and its identity confirmed by Mass Spectometry. Please refer to protocol tab for further experimental details.
  • Anti-NTF97/Importin beta antibody [3E9] (ab2811) at 1/5000 dilution + HeLa Whole cell extract

    Secondary
    HRP-conjugated anti mouse IgG at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size : 97 kDa

    Image and protocol courtesy of Rosamaria Mangiacasale and Patrizia Lavia, University 'La Sapienza' CNR, Italy

    HeLa whole cell extract was run on a 10%SDS-PAGE and transferred to PVDF membrane. Membrane was blocked for 30 mins in TBS/0.1% Tween/ 5% Milk; ab2811 (1/5000) was incubated for 1 hr in TBS/0.1%Tween/5% Milk and followed by 3 washes in TBS/ 0.1%Tween (3x 7 mins). Secondary antibody was incubated for 30 mins in a TBS/ 0.1% Tween solution.This was followed by 3 washes with the TBST solution (3x7 mins) and one wash in TBS. Western blot developed using ECL plus (Amersham).
  • Overlay histogram showing Jurkat cells stained with ab2811 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab2811, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

Anti-NTF97/Importin beta antibody [3E9] (ab2811)参考文献

This product has been referenced in:
  • Lee Y  et al. KPNB1 mediates PER/CRY nuclear translocation and circadian clock function. Elife 4:N/A (2015). IP, ICC/IF ; Mouse, Human . Read more (PubMed: 26319354) »
  • Germain MA  et al. Elucidating novel hepatitis C virus-host interactions using combined mass spectrometry and functional genomics approaches. Mol Cell Proteomics 13:184-203 (2014). Human . Read more (PubMed: 24169621) »

See all 20 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (NIH-3T3 cells)
Permeabilization No
Specification NIH-3T3 cells
Blocking step 1% Donkey Serum + 1% BSA + 0.1% Triton X-100, in PBS as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative Paraformaldehyde
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提交于 Nov 02 2015

Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing (10)
Sample Syrian Hamster Cell lysate - whole cell (BHK)
Specification BHK
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
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提交于 Oct 28 2013

Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing (10)
Sample Dog Cell lysate - whole cell (MDCK)
Specification MDCK
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
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提交于 Oct 28 2013

Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing (10)
Sample Cow Cell lysate - whole cell (MDBK)
Specification MDBK
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
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提交于 Oct 28 2013

Application Western blot
Loading amount 75 µg
Gel Running Conditions Reduced Denaturing (10)
Sample African Green Monkey Cell lysate - whole cell (COS cells)
Specification COS cells
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
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提交于 Oct 17 2013

Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing (10)
Sample Rat Cell lysate - whole cell (NRK cells)
Specification NRK cells
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
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提交于 Oct 17 2013

Application Western blot
Loading amount 75 µg
Gel Running Conditions Reduced Denaturing (10)
Sample Human Cell lysate - whole cell (HEK-293F and HeLa cells)
Specification HEK-293F and HeLa cells
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
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提交于 Oct 17 2013

Application Western blot
Loading amount 100 µg
Gel Running Conditions Reduced Denaturing (10)
Sample Mouse Tissue lysate - whole (Lung,Kidney & Spleen extracts)
Specification Lung,Kidney & Spleen extracts
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
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提交于 Oct 17 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Dog Cell lysate - whole cell (MDCK cells)
Loading amount 100 µg
Specification MDCK cells
Gel Running Conditions Reduced Denaturing (8% Tris-Glycine gel)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C
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提交于 May 14 2013

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Application Western blot
Sample Cow Cell lysate - whole cell (MDBK cells)
Loading amount 100 µg
Specification MDBK cells
Gel Running Conditions Reduced Denaturing (8% Tris-Glycine gel)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C
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提交于 May 13 2013

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