Overview

  • Product name
  • Description
    Goat polyclonal to KPNA4
  • Host species
    Goat
  • Tested applications
    Suitable for: IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Cow, Dog
  • Immunogen

    Peptide with sequence NSSANVPTEGFQF, from C Terminus of the protein sequence according to NP_002259

  • Positive control
    • Recombinant Human KPNA4 protein (ab114626) can be used as a positive control in WB. Human testis and ovary lysate. Human testis tissue sections and human small intestine sections.
  • General notes
    GenBank Accession Number – NP_002259.


    The nuclear import of karyophilic proteins is directed by short amino acid sequences termed nuclear localization signals (NLSs). Karyopherins, or importins, are cytoplasmic proteins that recognize NLSs and dock NLS-containing proteins to the nuclear pore complex. The protein encoded by this gene shares the sequence similarity with Xenopus importin-alpha and Saccharomyces cerevisiae Srp1. This protein is found to interact with the NLSs of DNA helicase Q1 and SV40 T antigen.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer
    pH: 7.30
    Preservative: 0.02% Sodium azide
    Constituents: Tris buffered saline, 0.5% BSA
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    Purified from goat serum by ammonium sulphate precipitation followed by antigen affinity chromatography using the immunizing peptide.
  • Primary antibody notes
    The nuclear import of karyophilic proteins is directed by short amino acid sequences termed nuclear localization signals (NLSs). Karyopherins, or importins, are cytoplasmic proteins that recognize NLSs and dock NLS-containing proteins to the nuclear pore complex. The protein encoded by this gene shares the sequence similarity with Xenopus importin-alpha and Saccharomyces cerevisiae Srp1. This protein is found to interact with the NLSs of DNA helicase Q1 and SV40 T antigen.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab6039 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P Use a concentration of 1 - 2 µg/ml.
WB Use a concentration of 0.2 - 1 µg/ml. Detects a band of approximately 55-60 kDa (predicted molecular weight: 58 kDa).Can be blocked with Human KPNA4 peptide (ab23144). A customer reported that milk can significantly suppress antigen-antibody interaction on nitrocellulose membrane in WB.

Target

  • Function
    Functions in nuclear protein import as an adapter protein for nuclear receptor KPNB1. Binds specifically and directly to substrates containing either a simple or bipartite NLS motif. 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. In vitro, mediates the nuclear import of human cytomegalovirus UL84 by recognizing a non-classical NLS. In vitro, mediates the nuclear import of human cytomegalovirus UL84 by recognizing a non-classical NLS.
  • Tissue specificity
    Highly expressed in testis, ovary, small intestine, heart, skeletal muscle, lung and pancreas, but barely detectable in kidney, thymus, colon and peripheral blood leukocytes.
  • Sequence similarities
    Belongs to the importin alpha family.
    Contains 10 ARM repeats.
    Contains 1 IBB domain.
  • Domain
    Consists of an N-terminal hydrophilic region, a hydrophobic central region composed of 10 repeats, and a short hydrophilic C-terminus. The N-terminal hydrophilic region contains the importin beta binding domain (IBB domain), which is sufficient for binding importin beta and essential for nuclear protein import.
    The IBB domain is thought to act as an intrasteric autoregulatory sequence by interacting with the internal autoinhibitory NLS. Binding of KPNB1 probably overlaps the internal NLS and contributes to a high affinity for cytoplasmic NLS-containing cargo substrates. After dissociation of the importin/substrate complex in the nucleus the internal autohibitory NLS contributes to a low affinity for nuclear NLS-containing proteins.
    The major and minor NLS binding sites are mainly involved in recognition of simple or bipartite NLS motifs. Structurally located within in a helical surface groove they contain several conserved Trp and Asn residues of the corresponding third helices (H3) of ARM repeats which mainly contribute to binding.
  • Cellular localization
    Cytoplasm. Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • FLJ31113 antibody
    • IMA3_HUMAN antibody
    • Importin alpha 3 antibody
    • Importin alpha 4 subunit antibody
    • Importin alpha Q1 antibody
    • Importin subunit alpha 3 antibody
    • Importin subunit alpha-3 antibody
    • IPO A3 antibody
    • IPOA 3 antibody
    • IPOA3 antibody
    • Karyopherin alpha 4 (importin alpha 3) antibody
    • Karyopherin alpha 4 antibody
    • Karyopherin alpha 4 importin alpha 3 antibody
    • Karyopherin alpha 4 subunit antibody
    • Karyopherin subunit alpha 4 antibody
    • Karyopherin subunit alpha-4 antibody
    • KPNA 4 antibody
    • Kpna4 antibody
    • MGC12217 antibody
    • MGC26703 antibody
    • OTTHUMP00000213880 antibody
    • QIP 1 antibody
    • Qip1 antibody
    • Qip1 protein antibody
    • SRP 3 antibody
    • SRP3 antibody
    see all

Images

  • All lanes : Anti-KPNA4 antibody (ab6039) at 1 µg/ml

    Lane 1 : Human testis
    Lane 2 : Human ovary
    Lane 3 : Mouse testis
    Lane 4 : Rat testis
    Lane 5 : Rat ovary

    Lysates/proteins at 35 µg per lane.

    Predicted band size: 58 kDa

  • Immunohistochemical analysis of 10% buffered formalin-fixed paraffin-embedded murine nephric tissue sections, labelling KPNA4 with ab6039 at  a dilution of 1/100 incubated for 12 hours at 4°C. Heat mediated antigen retrival was performed through 10mM citrate buffer at pH 6.0. Blocking was with 5% serum incubated for 1 hour at 21°C. Secondary was a polyclonal Donkey anti-Goat Alexa Fluor® 647 conjugate at 1/200. Counterstain is DAPI against nuclear DNA. The Negative control is the same as above, but in the abscence of the primary ab6039.

    See Abreview

  • ab6039 at 0.5µg/ml staining approximately 55-60kDa band of KPNA4 in Human Testis lysate by Western blot (ECL). ab6039 at 0.5µg/ml staining approximately 55-60kDa band of KPNA4 in Human Testis lysate by Western blot (ECL).
  • ab6039 (3µg/ml) staining Human Testis by IHC-P. Microwaved antigen retrieval with citrate buffer (pH 6), HRP-staining.
  • ab6039 at 5µg/ml staining KPNA4 in human small intestinal tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Steamed antigen retrieval with citrate buffer pH 6, AP-staining.

References

This product has been referenced in:
  • Zheng Y  et al. MicroRNA-24 inhibits the proliferation and migration of endothelial cells in patients with atherosclerosis by targeting importin-a3 and regulating inflammatory responses. Exp Ther Med 15:338-344 (2018). Read more (PubMed: 29250154) »
  • Major AT  et al. Development of a pipeline for automated, high-throughput analysis of paraspeckle proteins reveals specific roles for importin a proteins. Sci Rep 7:43323 (2017). Read more (PubMed: 28240251) »
See all 10 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM citrate buffer pH6
Permeabilization
No
Specification
kidney
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
10% buffered formalin

Abcam user community

Verified customer

Submitted Nov 11 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (pancreatic tumor)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM sodium citrate buffer pH6
Permeabilization
No
Specification
pancreatic tumor
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
10% buffered formalin

Abcam user community

Verified customer

Submitted Oct 29 2015

Answer

I hope that you are well.

I am sending this email to inquire about the updates you might have about the WB analysis. There were few anti KPNA4 products that were proving difficult to use and as per your last email you were suppose to contact me with the results.

I am wondering if these products are working now. Could you update me with any progress you have made? If you are still having problems then please let me know I will certainly try to find a solution of problem.

Many thanks for your cooperation in this case. I will be looking forward to hearing from you soon.

Read More

Question

Thank you very much for providing the replacement antibodies. We have now tested these in Western blotting and below is a summary of the results.


Anti-KPNA4 (ab6039)

This antibody produces a good signal in Western blot. However, this signal is still much weaker than the signal produced by anti-KPNA2 (ab6036) (used as a positive control) (see top panels inattached PowerPoint file). Interestingly, the signal is also weaker than the signal produced by the same antibody purchased fromAbcam in September 2006 (lower panels). Overall, although it might not be ideal, anti-KPNA4 (ab6039) clearly works and could be useful in our project.

Anti-KPNA6 (ab82149)

This antibody produces a doublet at the expected position forKPNAisoforms (between 50-75 kDa), suggesting cross-reaction with another KPNA. In addition, it produces multiple bands at higher mol. wt. some of these as strong as the bands at the expected position. Due to the cross-reaction and the overall low signal (compared to the positive control anti-KPNA2 (ab6036)) this antibody is unlikely to be useful for our project.

Anti-SRP1 (ab6035)

This antibody produces multiple bands of low intensity at positions higher than expected fromSRP1 (expected to run between 50-75 kDa). Overall, this antibody failed in Western blot and is of no use for our project.

In summary, we would like to test further antibodies forKPNA6 andSRP1. Could you please recommend any alternatives and provide them as replacements?

Thank you very much for your help and advice.

With regards,

Read More
Answer

Thank you for your email.

Two SRP1 antibodies failed, which is very strange. We have used SRP1 transfected 293T cells line in the characterization of antibody ab124646, this means that the 293T cells seems not expressing the protein. I would suggest using Hela cell line lysates as positive control.

KPAN6: please use 293 cell line lysates, human skeletal muscle lysates or brain lysates as positive control.

I am sure these antibodies will produce excellent results once correct cell line is used and with some protocol optimizations.

I hope this information will be helpful. Should you have any question please do not hesitate to ask.

Read More

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number 1092038.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HCT116, HT29 and Caco2 colon cancer cell lines)
Loading amount
30 µg
Specification
HCT116, HT29 and Caco2 colon cancer cell lines
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% in TBST

Abcam user community

Verified customer

Submitted Nov 07 2006

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up