Invitrogen

Importin beta-1 Monoclonal Antibody (3E9)

Advanced Verification

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated.

1 Published Figure

21 References

View all (23) KPNB1 antibodies

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Datasheet

Protocols

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Importin beta-1 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of Importin beta-1 using anti-Importin beta-1 monoclonal antibody (Product # MA3-070) shows staining in HMVEC Cells.

Importin beta-1 Antibody in Western Blot (WB) — Western blot was performed using Anti-Importin beta-1 Monoclonal Antibody (3E9) (Product # MA3-070) and a 97kDa band corresponding to Importin beta-1 was observed across 4 cell lines. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), Caco-2 (Lane 2), Hep G2 (Lane 3), NIH/3T3 (Lane 4) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody ( 1:5,000) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Reagent Kit (Product # WP20005).

Importin beta-1 Antibody in Flow Cytometry (Flow) — Flow cytometry analysis of Importin beta-1 in NIH-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 a Importin beta-1 monoclonal antibody (Product # MA3-070) at a dilution of 1 µg/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.

Importin beta-1 Antibody in Immunoprecipitation (IP) — Immunoprecipitation of KPNB1 was performed on K562 cells. Antigen-antibody complexes were formed by incubating approximately 500 µg whole cell lysate with 5 µg of KPNB1 monoclonal antibody (Product # MA3-070) rotating 60 min at RT. The immune complexes were captured on 625 µg of anti-mouse coated Dynabeads (Product # 11202D), washed extensively, and eluted with NuPAGE™ LDS Sample Buffer (Product # NP0007). Samples were resolved onto NuPAGE™ 4-12% Bis-Tris gel (Product # NP0335BOX). Lanes 1 and 3 are input and lanes 2 and 4 are IP. Proteins were transferred to PVDF membrane (Product # IB23001). Membrane was blocked in 5% milk. Target was detected using a KPNB1 monoclonal antibody (Product # MA3-070) at a dilution of 1:2000, followed by a 1:4000 dilution of secondary antibody. Chemiluminescent detection was performed using ECL Western Blotting Substrate (Product # 32106). Data courtesy of the Yeo lab as part of the ENCODE project (www.encodeproject.org).

Importin beta-1 Antibody in RNA Immunoprecipitation (RIP) — RNA immunoprecipitation (RIP) western of KPNB1 was performed on K562 cells. Antigen-antibody complexes were formed by incubating approximately 500 µg whole cell lysate with 5 µg of KPNB1 monoclonal antibody (Product # MA3-070) rotating 60 min at RT. The immune complexes were captured on 625 µg of anti-mouse coated Dynabeads (Product # 11202D), washed extensively, and eluted with NuPAGE™ LDS Sample Buffer (Product # NP0007). Samples were resolved onto NuPAGE™ 4-12% Bis-Tris gel (Product # NP0335BOX). Lanes 1 and 3 are input and lanes 2 and 4 are IP. Proteins were transferred to PVDF membrane (Product # IB23001). Membrane was blocked in 5% milk. Target was detected using a KPNB1 monoclonal antibody (Product # MA3-070) at a dilution of 1:2000, followed by a 1:4000 dilution of secondary antibody. Chemiluminescent detection was performed using ECL Western Blotting Substrate (Product # 32106). Data courtesy of the Yeo lab as part of the ENCODE project (www.encodeproject.org).

Importin beta-1 Antibody — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. HeLa cells were transfected with Importin beta-1 siRNA and decrease in signal intensity was observed in Western Blot application using Anti-Importin beta-1 Monoclonal Antibody (3E9) (Product # MA3-070). {KD}

Product Details

MA3-070

Applications	Tested Dilution	Publications
Western Blot (WB)	1:5,000	View 5 publications 5 publications
Immunocytochemistry (ICC/IF)	1:1,000	View 8 publications 8 publications
Flow Cytometry (Flow)	1 µg/test	-
Immunoprecipitation (IP)	Assay-dependent	View 3 publications 3 publications
Inhibition Assays (IA)	Assay-dependent	-
RNA Immunoprecipitation (RIP)	Assay-dependent	-
Neutralization (Neu)	-	View 3 publications 3 publications
ChIP assay (ChIP)	-	View 1 publication 1 publication
Immunohistochemistry (IHC)	-	View 1 publication 1 publication

Product Specifications
Species Reactivity	Bovine, Dog, Human, Mouse, Pig, Rat
Published species	Human, Mouse, Rat
Host/Isotype	Mouse / IgG2a
Class	Monoclonal
Type	Antibody
Clone	3E9
Immunogen	Purified NTF97 from bovine erythrocytes.
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Protein A
Storage buffer	PBS
Contains	0.05% sodium azide
Storage conditions	-20° C, Avoid Freeze/Thaw Cycles
RRID	AB_2133986

Product Specific Information

MA3-070 detects Importin beta 1 from human, canine, bovine, rat and mouse tissues as well as porcine cell lysates. Importin beta 1 is also known as NTF97, Nuclear Transport Factor p97, and karyopherin beta 1. This antibody does not detect bacterially expressed recombinant NTF97.

MA3-070 has been successfully used in Western blot, immunofluorescence, and immunoprecipitation procedures. By Western blot, this antibody detects a single ~97 kDa protein representing NTF97 in MDBK cell homogenate. Immunofluorescence staining of NTF97 in MDBK cells with MA3-070 results in the labeling of both the nuclear envelope (NE) and the cytoplasm and shows discontinuous, punctate staining along the rim of the NE. This antibody can inhibit import of nuclear localization sequence-containing proteins in permeabilized cells.

The MA3-070 antigen is purified NTF97 from bovine erythrocytes.

Target Information

Nucleocytoplasmic transport, a signal- and energy-dependent process, takes place through nuclear pore complexes embedded in the nuclear envelope. The import of proteins containing a nuclear localization signal requires the NLS import receptor, a heterodimer of importin alpha and beta subunits also known as karyopherins. Importin alpha binds the NLS-containing cargo in the cytoplasm and importin beta docks the complex at the cytoplasmic side of the nuclear pore complex. In the presence of nucleoside triphosphates and the small GTP binding protein Ran, the complex moves into the nuclear pore complex and the importin subunits dissociate. Importin alpha enters the nucleoplasm with its passenger protein and importin beta remains at the pore. Interactions between importin beta and the FG repeats of nucleoporins are essential in translocation through the pore complex. The protein encoded by this gene is a member of the importin beta family.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

O-GlcNAc modification of nuclear pore complexes accelerates bidirectional transport.

The Journal of cell biology

Yoo TY,Mitchison TJ

MA3-070 was used in Immunocytochemistry to develop high-throughput assays based on optogenetic probes to quantify the kinetics of nuclear import and export in living human cells.

Mon Jul 05 00:00:00 EDT 2021

Effect of Different Nuclear Localization Signals on the Subcellular Localization and Anti-HIV-1 Function of the MxB Protein.

Frontiers in microbiology

Chai K,Wang Z,Pan Q,Tan J,Qiao W,Liang C

MA3-070 was used in Immunocytochemistry to demonstrate that the bipartite nuclear localization signal has anti-HIV-1 activity when replacing the N-terminal sequence of the myxovirus resistance B gene.

Tue Jun 08 00:00:00 EDT 2021

HIV-1 resists MxB inhibition of viral Rev protein.

Emerging microbes & infections

Wang Z,Chai K,Liu Q,Yi DR,Pan Q,Huang Y,Tan J,Qiao W,Guo F,Cen S,Liang C

MA3-070 was used in Immunocytochemistry-immunoflourescence to report a new antiviral mechanism in which MxB restricts the nuclear import of HIV-1 regulatory protein Rev, and as a result, diminishes Rev-dependent expression of HIV-1 Gag protein.

Tue Dec 01 00:00:00 EST 2020

A potent CBP/p300-Snail interaction inhibitor suppresses tumor growth and metastasis in wild-type p53-expressing cancer.

Science advances

Li HM,Bi YR,Li Y,Fu R,Lv WC,Jiang N,Xu Y,Ren BX,Chen YD,Xie H,Wang S,Lu T,Wu ZQ

MA3-070 was used in Western Blotting to describe the inhibition of Snail by CYD19 in wild-type p53-expressing cancer.

Wed Apr 01 00:00:00 EDT 2020

Differential Roles of Cysteinyl Cathepsins in TGF-β Signaling and Tissue Fibrosis.

iScience

Zhang X,Zhou Y,Yu X,Huang Q,Fang W,Li J,Bonventre JV,Sukhova GK,Libby P,Shi GP

MA3-070 was used in Immunoprecipitation to demonstrate that TGF-β enhances CatS and CatK expression but reduces CatB and CatL expression in mouse kidney tubular epithelial cells.

Fri Sep 27 00:00:00 EDT 2019

Selenite targets eIF4E-binding protein-1 to inhibit translation initiation and induce the assembly of non-canonical stress granules.

Nucleic acids research

Fujimura K,Sasaki AT,Anderson P

MA3-070 was used in immunocytochemistry to study the role of 4EBP1 in the mechanism by which sodium selenite induces non-canonical stress granules and the implications for the chemotherapeutic use of selenite

Sat Sep 01 00:00:00 EDT 2012

Importin beta interacts with the endoplasmic reticulum-associated degradation machinery and promotes ubiquitination and degradation of mutant alpha1-antitrypsin.

The Journal of biological chemistry

Zhong Y,Wang Y,Yang H,Ballar P,Lee JG,Ye Y,Monteiro MJ,Fang S

MA3-070 was used in western blot to investigate the role of importin beta in intracellular protein degradation process

Fri Sep 30 00:00:00 EDT 2011

Localization of retinitis pigmentosa 2 to cilia is regulated by Importin beta2.

Journal of cell science

Hurd TW,Fan S,Margolis BL

MA3-070 was used in western blot to investigate the regulatory role of importin beta2 in the cilial localization of retinitis pigmentosa 2

Tue Mar 01 00:00:00 EST 2011

Nuclear entry of activated MAPK is restricted in primary ovarian and mammary epithelial cells.

PloS one

Smith ER,Cai KQ,Smedberg JL,Ribeiro MM,Rula ME,Slater C,Godwin AK,Xu XX

MA3-070 was used in western blot to study the alterations of MAPK activation and nuclear transport in primary ovarian and mammary epithelial cells

Thu Feb 18 00:00:00 EST 2010

Modulation of integrin-linked kinase nucleo-cytoplasmic shuttling by ILKAP and CRM1.

Cell cycle (Georgetown, Tex.)

Nakrieko KA,Vespa A,Mason D,Irvine TS,D'Souza SJ,Dagnino L

MA3-070 was used in immunocytochemistry to study the role of ILKAP and CRM-1 in modulating the movement of integrin-linked kinase between the nucleus and the cytoplasm

Tue Jul 15 00:00:00 EDT 2008

A novel mechanism of E2F1 regulation via nucleocytoplasmic shuttling: determinants of nuclear import and export.

Cell cycle (Georgetown, Tex.)

Ivanova IA,Vespa A,Dagnino L

MA3-070 was used in blocking or activating experiment to investigate the nucleocytoplasmic shuttling of E2F1

Sat Sep 01 00:00:00 EDT 2007

HuD distribution changes in response to heat shock but not neurotrophic stimulation.

The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society

Burry RW,Smith CL

Published figure using Importin beta-1 monoclonal antibody (Product # MA3-070) in Immunofluorescence

Sun Oct 01 00:00:00 EDT 2006

The N-terminal domain of the mammalian nucleoporin p62 interacts with other nucleoporins of the FXFG family during interphase.

Experimental cell research

Stochaj U,Bański P,Kodiha M,Matusiewicz N

MA3-070 was used in western blot to study the interaction of mammalian nucleoporin p62 interacts with the FXFG family protein during interphase

Tue Aug 01 00:00:00 EDT 2006

Multiple importins function as nuclear transport receptors for the Rev protein of human immunodeficiency virus type 1.

The Journal of biological chemistry

Arnold M,Nath A,Hauber J,Kehlenbach RH

MA3-070 was used in chromatin immunoprecipitation to study the binding of Rev to the nuclear transport receptors importins and its nuclear import through several transport pathways.

Fri Jul 28 00:00:00 EDT 2006

Enhanced intracellular mobility and nuclear accumulation of DNA plasmids associated with a karyophilic protein.

Human gene therapy

Mesika A,Kiss V,Brumfeld V,Ghosh G,Reich Z

MA3-070 was used in blocking or activating experiment to study the effect of nuclear localization signals on the intracellular transport of injected plasmid DNA

Tue Feb 01 00:00:00 EST 2005

Sugar-dependent nuclear import of glycosylated proteins in living cells.

Glycobiology

Rondanino C,Bousser MT,Monsigny M,Roche AC

MA3-070 was used in immunocytochemistry to compare the nuclear localization signal-mediated nuclear import pathway with the sugar-dependent nuclear import pathway

Tue Jul 01 00:00:00 EDT 2003

An immunohistochemical approach to monitor the prolactin-induced activation of the JAK2/STAT5 pathway in pancreatic islets of Langerhans.

The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society

Brelje TC,Svensson AM,Stout LE,Bhagroo NV,Sorenson RL

MA3-070 was used in immunohistochemistry to evaluate an immunohistochemical methodology for studying JAK2/STAT5 pathway in pancreatic islets of Langerhans

Fri Mar 01 00:00:00 EST 2002

Importin beta-mediated nuclear import of fibroblast growth factor receptor: role in cell proliferation.

The Journal of cell biology

Reilly JF,Maher PA

MA3-070 was used in immunoprecipitation to study the role of importin beta in the nuclear import of fibroblast growth factor receptor.

Mon Mar 19 00:00:00 EST 2001

Herpes simplex virus type 1 entry into host cells: reconstitution of capsid binding and uncoating at the nuclear pore complex in vitro.

Molecular and cellular biology

Ojala PM,Sodeik B,Ebersold MW,Kutay U,Helenius A

MA3-070 was used in immunocytochemistry to study the entry of herpes simplex type 1 virus into host cells.

Sat Jul 01 00:00:00 EDT 2000

Ran-independent nuclear import of cyclin B1-Cdc2 by importin beta.

Proceedings of the National Academy of Sciences of the United States of America

Takizawa CG,Weis K,Morgan DO

MA3-070 was used in immunoprecipitation to study the mechanism for the cyclin B1-Cdc2 import into the nucleus

Tue Jul 06 00:00:00 EDT 1999

Nucleocytoplasmic shuttling factors including Ran and CRM1 mediate nuclear export of NFAT In vitro.

The Journal of cell biology

Kehlenbach RH,Dickmanns A,Gerace L

MA3-070 was used in blocking/activating experiment to study the modulatory mechanism of the nuclear export of NFAT

Mon May 18 00:00:00 EDT 1998

Bioinformatics

Protein Aliases: HGNC:6400; importin 1; importin 90; Importin beta; importin beta-1 subunit; Importin subunit beta-1; Importin-90; KapB1; karyopherin (importin) beta 1; karyopherin beta 1; Karyopherin subunit beta-1; karyopherin,beta 1; MGC2155; MGC2156; MGC2157; Nuclear factor p97; Pore targeting complex 97 kDa subunit; PTAC97; SCG

Gene Aliases: AA409963; IMB1; Impnb; IPO1; IPOB; KPNB1; NTF97

UniProt ID: (Human) Q14974, (Rat) P52296, (Mouse) P70168

Entrez Gene ID: (Bovine) 507804, (Dog) 491042, (Pig) 100518449, (Human) 3837, (Rat) 24917, (Mouse) 16211

Molecular Function: transporter

Function(s)

protein import into nucleus, docking protein import into nucleus, translocation apoptotic DNA fragmentation protein import into nucleus NLS-bearing protein import into nucleus ribosomal protein import into nucleus mitotic chromosome movement towards spindle pole mitotic metaphase plate congression modulation by virus of host process astral microtubule organization Ran protein signal transduction establishment of mitotic spindle localization establishment of protein localization intracellular transport of virus mitotic spindle assembly intracellular protein transport nucleocytoplasmic transport transport protein transport

Process(es)

protein binding nuclear localization sequence binding zinc ion binding Ran GTPase binding protein transporter activity enzyme binding protein domain specific binding poly(A) RNA binding Hsp90 protein binding kinesin binding protein complex binding

Importin beta-1 Monoclonal Antibody (3E9)

Product Details

MA3-070

Western Blot (WB)

Immunocytochemistry (ICC/IF)

Flow Cytometry (Flow)

Immunoprecipitation (IP)

Inhibition Assays (IA)

RNA Immunoprecipitation (RIP)

Neutralization (Neu)

ChIP assay (ChIP)

Immunohistochemistry (IHC)

Species Reactivity

Published species

Host/Isotype

Class

Type

Clone

Immunogen

Conjugate

Form

Concentration

Purification

Storage buffer

Contains

Storage conditions

RRID

Product Specific Information

Target Information

Bioinformatics

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