Overview

  • Product name

    Anti-KIRREL antibody
  • Description

    Rabbit polyclonal to KIRREL
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ELISA, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat, Rabbit, Horse, Chicken, Guinea pig, Cow, Cat, Dog, Pig
  • Immunogen

    Synthetic peptide corresponding to a region within internal sequence (FLEVGTLERY TVERTNSGSG VLSTLTINNV MEADFQTHYN CTAWNSFGPG) of Human KIRREL (NP_060710).(Peptide available as ab109278.)

  • Positive control

    • HepG2 cell lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab82804 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: 66 kDa.Can be blocked with KIRREL peptide (ab109278). Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
ELISA Use at an assay dependent concentration.
IHC-P Use a concentration of 10 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

  • Function

    Plays a significant role in the normal development and function of the glomerular permeability. Signaling protein that needs the presence of TEC kinases to fully trans-activate the transcription factor AP-1.
  • Tissue specificity

    Abundantly expressed in kidney. Specifically expressed in podocytes of kidney glomeruli.
  • Sequence similarities

    Belongs to the immunoglobulin superfamily.
    Contains 5 Ig-like C2-type (immunoglobulin-like) domains.
  • Post-translational
    modifications

    Phosphorylation probably regulates the interaction with NSH2.
    N-glycosylated.
  • Cellular localization

    Cell membrane. Predominantly located at podocyte slit diaphragm.
  • Information by UniProt
  • Database links

  • Alternative names

    • FLJ10845 antibody
    • Kin of IRRE like (Drosophila) antibody
    • Kin of IRRE like antibody
    • Kin of IRRE like protein 1 antibody
    • Kin of IRRE-like protein 1 antibody
    • Kin of irregular chiasm like protein 1 antibody
    • Kin of irregular chiasm-like protein 1 antibody
    • KIRR1_HUMAN antibody
    • Kirrel antibody
    • MGC129542 antibody
    • MGC129543 antibody
    • NEPH 1 antibody
    • NEPH1 antibody
    • Nephrin like protein 1 antibody
    • Nephrin related antibody
    • Nephrin-like protein 1 antibody
    • OTTHUMP00000024055 antibody
    see all

Images

  • Anti-KIRREL antibody (ab82804) at 1 µg/ml (in 5% skim milk / PBS buffer) + HepG2 cell lysate at 10 µg

    Secondary
    HRP conjugated anti-Rabbit IgG at 1/50000 dilution

    Predicted band size: 66 kDa
    Observed band size: 64 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 40 kDa. We are unsure as to the identity of these extra bands.



    Gel concentration: 12%
  • IHC image of ab82804 staining in Human normal kidney formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab82804, 10µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

References

This product has been referenced in:

  • Zhang MJ  et al. Overexpression of Kin of IRRE-Like Protein 1 (KIRREL) in Gastric Cancer and Its Clinical Prognostic Significance. Med Sci Monit 24:2711-2719 (2018). Read more (PubMed: 29717104) »
  • Durcan PJ  et al. Identification and characterization of novel Kirrel isoform during myogenesis. Physiol Rep 1:e00044 (2013). Read more (PubMed: 24303129) »
See all 3 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Answer

Thank you very much for your call today and for letting us know about the trouble with this antibody.

As we discussed, I'm sending the blocking peptide free of charge on the order ***, which should arrive next week. Please keep me updated about the results of the blocking assay and with the positive control kidney lysate.

Here is our protocol for the blocking assay that we discussed-

https://www.abcam.com/index.html?pageconfig=resource&rid=11378

I look forward to hearing from you. If you have further questions or need anything else, please let me know and I'll be happy to help.

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Question

Thank you so much as well for sending the WB images. I do have a couple more questions for you to understand your experiment better: 1) Have you tried - besides mouse and rat samples - also human samples? The antibody has been tested so far only on human samples and we do not know if and how well it wouild work on mouse or rat samples. Mouse and rat is only predicted to react based on sequence homology. For instance, I am not certain why one would see three bands in the well-working lots for mouse (middle image in the ppt-slide): 45, 70 and 100 kDa? The mouse protein comes in 2 isoforms with MW of 87 and 70 kDa. The protein can be glycosylated which could shift the MW to larger number - depending on the extent of glycosylation. Thus, I am not quite certain if the antibody works well for mouse samples in general. ANSWER: I have not yet tried the antibody on human samples. The band at 45 kd in the mouse liver I am unsure about it disappears with the use of a blocking peptide. The other bands Im confident are kirrel transcripts. I have preformed immunopreciptation and deglycosylation studies using pngase F with previous aliquots of the kirrel antibody and band redcutions are present. however with the antibody which I have reported that I have a problem with I will be unable to use it for immunopreciptation experimetns due to the presence of the unkonwn band that is also in the protein marker lane which has not shown up in any of my previous aliquots. 2) At which MW did you see bands in the other mouse samples when the lots worked well for you? ANSWER: The size of bands depends on the specific tissue types/cell types. via deglycosylation studies the bands present are dependent on the cel type so bands can be present ranging from 80kd to 120kd. Also lower bands can be prsent rangin from 70-75kd approx also due to glyscosylation. 3) What does the literature report about the MW of the protein in mouse samples? ANSWER: Majority of litreature reports kirrel to be present at approximate 110kd. Majority of work done in kidney. Kirrel transmembrane protein. therefore heavily posttranslationaly modified is cell type dependent. the kirrel antibody only recognises a band of approximately 85kd for example in mouse kidney in contrast to majority of published litreature however there is one report of a kirre band at 90 kd in murine/rodent kidney. I have detected a band of 120kd in mouse brain along with other bands likely due to glycosylation however have to confirm this is brain.. 4) In which of your samples (mouse liver, kidney, heart, spleen, testis and skeletal muscle tissue and rat) would you expect to see the protein expressed at detectable levels and at which MW? ANSWER: This question is not easy to answer for all of the reasons mentioned above it depends what is the epitope of the antibody for example the kirrel antibody epitope spans a putative glycosylation site. It has been demonstrated that presence/lack of glycosylated resideues can inhibit antibody binding. 5) The Femto Supersignal detection kit is very sensistive and can lead to non-specific bands. I would suggest trying the Pico kit instead to reduce high background. ANSWER: This in my opinon is not a good suggestion for example if in some cell types express the protein is expressed at low levels using the pico kit may not be able to detect the protein and i get a false negative result. To conclue my problem is that the antibody is clearly giving non specific binding on my membranes which was not present in other aliquots.

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Answer

Thank you so much for your email and the additional information. I think the best way to proceed is the following: Mouse, rat samples and IP have all not been tested with this antibody and are therefore not covered under our guarantee. But since previous lots seem to have worked for you, it would be great to obtain this information in form of an Abreview. What we usually offer our customers who use antibodies in untested applications and/or species is our testing discount. Thus, if you please could submit an Abreview(s) for your previous mouse and/or rat and/or IP data, the discount code I am sending you below will become active and it can be applied to your next order (for instance another vial/lot of ab82804). The discount is for 100% of the price of the antibody you are testing (which would be ab82804). In addition, for each submitted Abreview, you will receive points that can also be applied to a future order. Once we receive your Abreview and publish it, we will be able to add mouse/rat/IP as tested species/application to the datasheet, and going forward it will be covered under our Abromise guarantee (https://www.abcam.com/abpromise) and can be replaced or refunded. Here is the discount code and additional information about the testing discount program: DISCOUNT CODE: xxx Expiration date: xxx I am very pleased to hear you would like to accept our offer and test ab82804 in mouse/rat/IP. This code will give you 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for mouse/rat/IP and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount. If you have any further questions, please let me know. I'd be happy to help.

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