Overview

  • Product name
  • Description
    Rabbit polyclonal to LRPPRC
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Rat
  • Immunogen

    Recombinant fragment corresponding to a region within amino acids 611-907 of Human LRPPRC (NP_573566)

  • Positive control
    • H1299, HeLa, 293T, A431, HepG2, Molt-4 and Raji cell lines. SW480 Xenograft.

Properties

Applications

Our Abpromise guarantee covers the use of ab97505 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 1/500 - 1/3000. Predicted molecular weight: 158 kDa.
IHC-P 1/100 - 1/500.
ICC/IF 1/100 - 1/200.

Target

  • Function
    May play a role in RNA metabolism in both nuclei and mitochondria. In the nucleus binds to HNRPA1-associated poly(A) mRNAs and is part of nmRNP complexes at late stages of mRNA maturation which are possibly associated with nuclear mRNA export. May bind mature mRNA in the nucleus outer membrane. In mitochondria binds to poly(A) mRNA. Plays a role in translation or stability of mitochondrially encoded cytochrome c oxidase (COX) subunits. May be involved in transcription regulation. Cooperates with PPARGC1A to regulate certain mitochondrially encoded genes and gluconeogenic genes and may regulate docking of PPARGC1A to transcription factors. Seems to be involved in the transcription regulation of the multidrug-related genes MDR1 and MVP. Part of a nuclear factor that binds to the invMED1 element of MDR1 and MVP gene promoters. Binds single-stranded DNA.
  • Tissue specificity
    Expressed ubiquitously. Expression is highest in heart, skeletal muscle, kidney and liver, intermediate in brain, non-mucosal colon, spleen and placenta, and lowest in small intestine, thymus, lung and peripheral blood leukocytes.
  • Involvement in disease
    Leigh syndrome French-Canadian type
  • Sequence similarities
    Contains 20 PPR (pentatricopeptide) repeats.
  • Cellular localization
    Mitochondrion. Nucleus, nucleoplasm. Nucleus inner membrane. Nucleus outer membrane. Seems to be predominantly mitochondrial.
  • Information by UniProt
  • Database links
  • Alternative names
    • 130 kDa leucine-rich protein antibody
    • gp130 antibody
    • Leucine-rich PPR motif-containing protein antibody
    • Leucine-rich PPR-motif containing protein antibody
    • LPPRC_HUMAN antibody
    • LRP 130 antibody
    • lrpprc antibody
    • mitochondrial antibody
    see all

Images

  • All lanes : Anti-LRPPRC antibody (ab97505) at 1/1000 dilution

    Lane 1 : H1299 whole cell lysate
    Lane 2 : HeLa whole cell lysate

    Lysates/proteins at 30 µg per lane.

    Predicted band size: 158 kDa



    5% SDS-PAGE
  • Immunofluorescence analysis of paraformaldehyde-fixed HeLa, using ab97505 at 1/200 dilution. Lower panel is also merged with DNA probe.
  • Immunohistochemical analysis of paraffin-embedded SW480 Xenograft, using ab97505 at 1/100 dilution.

References

This product has been referenced in:
  • Sacoman JL  et al. Mitochondrial O-GlcNAc Transferase (mOGT) Regulates Mitochondrial Structure, Function, and Survival in HeLa Cells. J Biol Chem 292:4499-4518 (2017). Read more (PubMed: 28100784) »
  • Edgett BA  et al. The effect of acute and chronic sprint-interval training on LRP130, SIRT3, and PGC-1a expression in human skeletal muscle. Physiol Rep 4:N/A (2016). WB . Read more (PubMed: 27604398) »
See all 4 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Application
Western blot
Sample
Human Tissue lysate - whole (Skeletal Muscle)
Gel Running Conditions
Reduced Denaturing (6% Gel)
Loading amount
3.25 µg
Specification
Skeletal Muscle
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Nov 26 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.

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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,

Read More
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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