Anti-KPNB1 antibody [3E9]

• 产品名称

  Anti-KPNB1抗体[3E9]
  参阅全部 KPNB1 一抗

• 描述

  小鼠单克隆抗体[3E9] to KPNB1

• 宿主

  Mouse

• 经测试应用

  适用于: Flow Cyt, IP, WB, ICC/IFmore details

• 种属反应性

  与反应: Mouse, Rat, Human
  

• 免疫原

  Full length native protein (purified) corresponding to Cow KPNB1. Purified from Bovine erythrocytes.
  Database link: E1BFV0

• 阳性对照

• WB: HeLa, Caco-2, NIH/3T3, U251 MG, HepG2 and C6 whole cell lysates. ICC: HMVEC cells, PTK cells. Flow Cyt: PC-12, Jurkat and NIH 3T3 cells.

• 常规说明

  
  

  The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

  If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

• 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

• 浓度

• 100 µl 浓度为 1 mg/ml

• 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

The Abpromise guarantee

Abpromise™承诺保证使用ab2811于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

• 数据库链接

  hide

• Unigene: 706168 Human

• Unigene: 251013 Mouse

• Unigene: 11061 Rat

• Entrez Gene: 3837 Human

• Entrez Gene: 16211 Mouse

• Entrez Gene: 24917 Rat

• Omim: 602738 Human

• SwissProt: Q14974 Human

• SwissProt: P70168 Mouse

• SwissProt: P52296 Rat

• Unigene: 532793 Human

• 别名

• 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

• NTF97/Importin beta antibody

• Nuclear factor P97 antibody

• Pore targeting complex 97 kDa subunit antibody

• PTAC97 antibody

• IMB 1 antibody

• IMB1 antibody

• IMB1_HUMAN antibody

Western blot - Anti-KPNB1 antibody [3E9] (ab2811)

All lanes : Anti-KPNB1 antibody [3E9] (ab2811) at 1/5000 dilution

Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : Caco-2 (human colorectal adenocarcinoma cell line) whole cell lysate
Lane 3 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 4 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate

Lysates/proteins at 30 µg per lane.

Secondary
All lanes : Goat anti-Mouse IgG H+L (HRP) at 1/4000 dilution

Developed using the ECL technique.

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

• 

  Immunocytochemistry/ Immunofluorescence - Anti-KPNB1 antibody [3E9] (ab2811)

  Immunofluorecent analysis of HMVEC (Human Lung Microvascular Endothelial cells) cells stained for KPNB1 using ab2811.

Immunocytochemistry/ Immunofluorescence - Anti-KPNB1 antibody [3E9] (ab2811)This image is courtesy of Roberto Giambruno, Marilena Ciciarello and Patrizia Lavia

NIH/3T3 (Mouse embryo fibroblast cell line) 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 - Anti-KPNB1 antibody [3E9] (ab2811)Image and protocol courtesy of Rosamaria Mangiacasale and Patrizia Lavia, University 'La Sapienza' CNR, Italy

  Immunoprecipitation of Importin beta, in HeLa (Human epithelial cell line from cervix adenocarcinoma) 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.

• 

  Flow Cytometry - Anti-KPNB1 antibody [3E9] (ab2811)

  Flow cytometry analysis of KPNB1 in Jurkat (Human T cell leukemia cell line from peripheral blood) 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.

Western blot - Anti-KPNB1 antibody [3E9] (ab2811)

All lanes : Anti-KPNB1 antibody [3E9] (ab2811) at 1/50 dilution

Lane 1 : U-251 MG (Human brain glioma cell line) whole cell lysate
Lane 2 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 3 : C6 (Rat glial tumor cell line) whole cell lysate

Lysates/proteins at 25 µg per lane.

Secondary
All lanes : HRP-conjugated secondary antibody

Predicted band size: 97 kDa
Observed band size: 100 kDawhy is the actual band size different from the predicted?

• 

  Western blot - Anti-KPNB1 antibody [3E9] (ab2811)This image is courtesy of an anonymous Abreview

  All lanes : Anti-KPNB1 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
  All lanes : 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
  

  See Abreview

• 

  Western blot - Anti-KPNB1 antibody [3E9] (ab2811)Image and protocol courtesy of Rosamaria Mangiacasale and Patrizia Lavia, University 'La Sapienza' CNR, Italy

  Anti-KPNB1 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
  
  
  
  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).

• 

  Immunocytochemistry/ Immunofluorescence - Anti-KPNB1 antibody [3E9] (ab2811)

  Immunolocalization of KPNB1 in PTK (Long-nosed potoroo epithelial kidney cell line) cells using ab2811.

• 

  Flow Cytometry - Anti-KPNB1 antibody [3E9] (ab2811)

  Flow cytometry analysis of KPNB1 in PC-12 (Rat adrenal gland pheochromocytoma cell line) 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 - Anti-KPNB1 antibody [3E9] (ab2811)

  Flow cytometry analysis of KPNB1 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 - Anti-KPNB1 antibody [3E9] (ab2811)

  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.