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Human/Mouse/Rat Importin beta/KPNB1 Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB8209

R&D Systems, part of Bio-Techne
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MAB8209
MAB8209-SP

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Mouse

Applications

Validated:

Immunocytochemistry, Western Blot

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 845208

Product Specifications

Immunogen

E. coli-derived recombinant human Importin beta/KPNB1
Met1-Gly155
Accession # Q14974

Specificity

Detects human Importin beta/KPNB1 in ELISA and Western Blot. It detects mouse and rat Importin beta/KPNB1 in Western Blots.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Scientific Data Images for Human/Mouse/Rat Importin beta/KPNB1 Antibody

Detection of Human, Mouse, and Rat Importin  beta/KPNB1 antibody by Western Blot.

Detection of Human, Mouse, and Rat Importin beta/KPNB1 by Western Blot.

Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, NIH-3T3 mouse embryonic fibroblast cell line, and Rat-2 rat embryonic fibroblast cell line. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Importin beta/KPNB1 Monoclonal Antibody (Catalog # MAB8209) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Importin beta/KPNB1 at approximately 97 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Importin  beta/KPNB1 antibody in HeLa Human Cell Line by Immunocytochemistry (ICC).

Importin beta/KPNB1 in HeLa Human Cell Line.

Importin beta/KPNB1 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Mouse Anti-Human Importin beta/KPNB1 Monoclonal Antibody (Catalog # MAB8209) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Human Importin beta/KPNB1 by Western Blot

Detection of Human Importin beta/KPNB1 by Western Blot

cNLSs are necessary for Gli2 nuclear import.(A) Sequences in Gli2_Mm predicted as cNLSs by cNLS-Mapper [40] using a cut-off of 5 with the corresponding scores (out of 10). Residues that were mutated for alanines in the mutNLS constructs are in bold. The mutated sequences are not predicted (n.p.) as cNLS. (B) Transduced NIH/3T3 cells expressing GFP-Gli2, wt or cNLS mutants (mutNLS-1, mutNLS-2 or mutNLS-1+2), were treated with SAG for 90 min or DMSO as control. Cells were stained for cilium (anti-Ac.Tub, red), GFP-Gli2 (anti-GFP, green) and nucleus (DAPI, blue). The same pictures are also shown with the channel corresponding to GFP-Gli2 in grey scale to help the visualization. Scale bar: 10 μm. (C) Quantification of nuclear Gli2 was performed as explained in Fig 1C. Black, solid line indicates comparison between control and Hh activated conditions for wtGli2, dotted line indicates comparison between wtGli2 under basal condition and mutNLS-1 or mutNLS-1+2 under basal or activated conditions and black, discontinued line indicates comparison between activated conditions for wtGl2 and mutNLS-2. *p<0.01, *** p<0.0001 (Kruskal-Wallis test). (B) and (C) are representative of four independent experiments and at least 70 cells were analysed for each condition. (D) WB detecting Imp-alpha 1 after precipitating GFP-Gli2 or GFP-Gli2-mutNLS-1+2 with GFP-Trap from NIH/3T3 Flp-In expressing at endogenous levels the constructs mentioned above. Membranes were cut at different levels so as to detect in the same samples the precipitated GFP-Gli2 (using an anti-Gli2 antibody) and GFP (left panels). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/27579771), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human/Mouse/Rat Importin beta/KPNB1 Antibody

Application
Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell line

Western Blot

1 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line, NIH‑3T3 mouse embryonic fibroblast cell line, and Rat‑2 rat embryonic fibroblast cell line

Reviewed Applications

Read 3 reviews rated 4.7 using MAB8209 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Importin beta/KPNB1

KPNB1 (Karyopherin subunit beta 1), also called Importin beta1, NTF97 (nuclear transport factor, 97 kDa) or PTAC97 (pore targeting complex, 97 kDa), is a member of the Importin b family of proteins. It is ubiquitously expressed and plays many roles in the nucleus. Primarily, KPNB1 transports various complexes from cytoplasm into nucleus, either alone or with a KPNA adaptor. The 876 amino acid (aa) human KPNB1 contains an N-terminal IBB/Importin b domain (aa 21-101), and eight HEAT (solenoid) domains (aa 124-726). A 731 aa splice variant begins with Met146, while an 845 aa variant shows a 2 aa substitution for aa 702-734. Binding sites for adaptors and for its regulator, Ran-GTP, overlap within the HEAT domains. Human KPNB1 aa 1-155 shares 99% and 97% aa sequence identity with mouse and rat KPNB1, respectively.

Long Name

Karyopherin beta 1

Alternate Names

IMB1, Impnb, Importin 1, IPOB, KPNB1, NTF97, PTAC97

Entrez Gene IDs

3837 (Human); 16211 (Mouse); 24917 (Rat)

Gene Symbol

KPNB1

UniProt

Additional Importin beta/KPNB1 Products

Product Documents for Human/Mouse/Rat Importin beta/KPNB1 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human/Mouse/Rat Importin beta/KPNB1 Antibody

For research use only

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