Key features and details
- Mouse monoclonal [3E9] to KPNB1
- Suitable for: Flow Cyt, IP, WB, ICC/IF
- Reacts with: Mouse, Rat, Human
- Isotype: IgG2a
製品名Anti-KPNB1 antibody [3E9]
KPNB1 一次抗体 製品一覧
製品の詳細Mouse monoclonal [3E9] to KPNB1
アプリケーション適用あり: Flow Cyt, IP, WB, ICC/IFmore details
種交差性交差種: Mouse, Rat, Human
Full length native protein (purified) corresponding to Cow KPNB1. Purified from Bovine erythrocytes.
- WB: U251 MG, HepG2 and C6 whole cell lysates. ICC: HMVEC cells, PTK cells. Flow Cyt: PC-12, Jurkat and NIH 3T3 cells.
Previously labelled as NTF97/Importin beta.
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保存方法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.
バッファーPreservative: 0.05% Sodium azide
Concentration information loading...
精製度Protein A purified
一次抗体 備考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.
ChIP Related Products
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.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
|IP||Use at an assay dependent concentration.|
|WB||1/5000. Detects a band of approximately 97 kDa (predicted molecular weight: 97 kDa).|
機能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.
配列類似性Belongs to the importin beta family.
Contains 8 HEAT repeats.
Contains 1 importin N-terminal domain.
細胞内局在Cytoplasm. Nucleus envelope.
- Information by UniProt
- IMB 1 antibody
- IMB1 antibody
- IMB1_HUMAN antibody
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.
All lanes : HRP-conjugated secondary antibody
Predicted band size: 97 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?
Immunofluorecent analysis of HMVEC (Human Lung Microvascular Endothelial cells) cells stained for KPNB1 using ab2811.
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 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 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.
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
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
Anti-KPNB1 antibody [3E9] (ab2811) at 1/5000 dilution + HeLa Whole cell extract
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).
Immunolocalization of KPNB1 in PTK (Long-nosed potoroo epithelial kidney cell line) cells using 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 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.
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.
ab2811 は 46 報の論文で使用されています。
- Huber S et al. Recruitment of Host Nuclear Pore Components to the Vicinity of Theileria Schizonts. mSphere 5:N/A (2020). PubMed: 32024710
- Hazawa M et al. Karyopherin-ß1 Regulates Radioresistance and Radiation-Increased Programmed Death-Ligand 1 Expression in Human Head and Neck Squamous Cell Carcinoma Cell Lines. Cancers (Basel) 12:N/A (2020). PubMed: 32276424
- Oostdyk LT et al. An epilepsy-associated mutation in the nuclear import receptor KPNA7 reduces nuclear localization signal binding. Sci Rep 10:4844 (2020). PubMed: 32179771
- Oostdyk LT et al. Characterization of the Importin-ß binding domain in nuclear import receptor KPNA7. Biochem J 476:3413-3434 (2019). PubMed: 31642884
- Guo L et al. Phosphorylation of importin-a1 by CDK1-cyclin B1 controls mitotic spindle assembly. J Cell Sci 132:N/A (2019). PubMed: 31434716