Overview

  • Product name
  • Description
    Rabbit polyclonal to KPNA4
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WBmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat, Rabbit, Horse, Chicken, Guinea pig, Cow, Cat, Dog
  • Immunogen

    Synthetic peptide corresponding to a region within N terminal amino acids 36 - 85 (LRKNKRDEHL LKRRNVPHED ICEDSDIDGD YRVQNTSLEA IVQNASSDNQ) of KPNA4 (NP_002259).

  • Positive control
    • HeLa cell lysate.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer
    Preservative: 0.09% Sodium azide
    Constituents: 2% Sucrose, PBS
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    ab81451 is purified by a peptide affinity chromatography method.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab81451 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
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 58 kDa. Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.

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

  • Western blot analysis of HeLa cell lysate labeling KPNA4 with ab81451 at 0.2-1 µg/ml.
  • Western blot analysis of MCF7 cell lysate labeling KPNA4 with ab81451 at 1.0µg/ml.
  • Western blot analysis of Human fetal liver tissue lysate labeling KPNA4 with ab81451 at 1.0µg/ml.

References

ab81451 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

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.

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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.

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Question

Thank you for your valuable advice you sent recently to Jane Sharps regarding antibodies.
Just to clarify your last point, we do use293T celllysates and analyse endogenous proteins. Unfortunately, we cannot use over-expression of tagged proteins inour project.
I fully agree with you that by optimizing conditions(e.g. antibody concentrations, incubation times, etc) it could be possible to improve the signals these antibodies produce. However, in my experience, such modificationsgive some improvement, but are unlikely to produce a major effect.Weneed to be able to detect proteins at much lower concentrations than weused in the experiments to test their activity (see enclosed information).I am afraid, for the purpose of our project these antibodies are unlikely to be useful, although I agree with you that they could beused for detecting large amounts of proteins, especially if they are over-expressed.
For the purpose of our project we need antibodies that are able to detect much lower quantities of proteins. In this respect, your antibodies forKPNA2 (ab6036) andKPNA3 (ab6038) (see enclosed information) we purchased earlier are ideal. We are seeking antibodies of similar quality for KPNA1, KPNA4,and KPNA6.
I am aware ofAbcam selling a range of antibodies for each of these proteins. Would you be able to replacethe KPNA1, KPNA4, andKPNA6 antibodies we have purchased with ones which would produce signals similar to that ofKPNA2 (ab6036) andKPNA3 (ab6038) in Western blot (an ideally also work in immunoprecipitations).
Your advice would be much appreciated.
With regards,

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Answer

Thank you for contacting us.

We have 3 antibodies ab6035, ab6039 and ab82149 available from the same collaborator as for ab6038, and ab6036. It is hard to guess if these 3 antibodies will perform as good as ab6038/ ab6036 antibodies or even better than those. The only way to check this is trying the antibodies.

Please check the datasheets and let me know if you are interested in these antibodies; I will then send these asap.
Please also do not forget to publish abreviews. We provide 220 (100 abreview + 120 image) abpoints per Abreview, these can be converted into Amzone vouchers or money off in the next purchase.

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Answer

Thank you very much for filling the questionnaires. I am glad to hear that the antibody ab97505 is working in retest experiment. Because you have not mentioned ab88859 so I am assuming that this antibody is also successful.

I have checked the protocol and images you have kindly provided and I can say these antibodies will produce excellent results with some general trouble shooting. My suggestions are as follows;

ab105346, This antibody produced correct band however visible at 15 min exposure rather than 1 minutes. It seems the results can be improved

All the antibodies are used at 1/400 dilution which is OK but because the immune response of rabbit is different with different antigens so we always recommend our customer optimizing the conditions with individual antibodies. I would suggest trying the antibody at 1/300 - 1/200 dilution if same amount of lysates is being used.
Overnight incubation of primary at 4C is recommended.
1/50 of total lysates, this actually does not tell how much protein was loaded. I would suggest determining the protein concentration of lysates by Lowry or Bradford method and then loading 30-50 ug protein per well. It may well be that the lysates concentration is not enough.
In our own experience TBST produces good results. Try 4 washes, 10 minutes first and 5 minutes 3 washes are recommend.
Lower dilution of secondary can also be tried.



ab81451

This antibody produced correct band size, which I agree is faint. So the results can be improved by using high concentration of protein and lower dilution of antibody.
There are black dots in the image, you may need to filter the blocking and antibody dilution buffer.
We recommend loading 30-50 of protein
Other similar recommendations of ab105346



ab105350

Normally the KPNA6 antibodies gives band 60-65 kDa. I have checked few publications where people have observed 75 kDa band however with GST- KPNA6.
Other similar recommendations of ab105346



Finally, In the original email you have written 293T over expressed lysates are being used; could you confirm if all of these proteins are over expressed? Is the fragment tagged e.g. myc or GST. This may also explain the 75 kDa which may be higher due to tag.

I am sure the results will improve with these changes. If there is no improvement, let me know I will replace the antibodies.

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Answer

Thank you for calling us and for alerting us to the problem you are experiencing with our product. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

As we discussed on the phone, I amattaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.

I look forward to receiving your reply.

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

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