Overview

  • Product name
  • Description
    Rabbit polyclonal to Nephrin
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    A 14 amino acid peptide from near the carboxy terminus of human Nephrin.

  • Positive control
    • WB: HEK293 and Mouse kidney lysate. IHC: Mouse kidney tissue.
  • General notes
    Although there are publications stating this antibody works with Rat species in WB (PMID 19052104). The antibody however fail to stain the Rat kidney sections hence we have removed the species(from customer feedback). Further feedback using this antibody with Rat tissues would be very welcome

Properties

Applications

Our Abpromise guarantee covers the use of ab58968 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 5 - 10 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB Use a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 85 kDa (predicted molecular weight: 138 kDa).

Target

  • Function
    Seems to play a role in the development or function of the kidney glomerular filtration barrier. Regulates glomerular vascular permeability. May anchor the podocyte slit diaphragm to the actin cytoskeleton. Plays a role in skeletal muscle formation through regulation of myoblast fusion.
  • Tissue specificity
    Specifically expressed in podocytes of kidney glomeruli.
  • Involvement in disease
    Defects in NPHS1 are the cause of nephrotic syndrome type 1 (NPHS1) [MIM:256300]; also known as Finnish congenital nephrosis (CNF). A renal disease characterized clinically by proteinuria, hypoalbuminemia, hyperlipidemia, and edema. Kidney biopsies show non-specific histologic changes such as focal segmental glomerulosclerosis and diffuse mesangial proliferation. Some affected individuals have an inherited steroid-resistant form and progress to end-stage renal failure.
  • Sequence similarities
    Belongs to the immunoglobulin superfamily.
    Contains 1 fibronectin type-III domain.
    Contains 8 Ig-like C2-type (immunoglobulin-like) domains.
  • Developmental stage
    In 23-week-old embryo found in epithelial podocytes of the periphery of mature and developing glomeruli.
  • Post-translational
    modifications
    Phosphorylated on tyrosine residues.
  • Cellular localization
    Cell membrane. Predominantly located at podocyte slit diaphragm between podocyte foot processes. Also associated with podocyte apical plasma membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • CNF antibody
    • Nephrin antibody
    • Nephrosis 1 congenital Finnish type antibody
    • Nephrosis 1, congenital, Finnish type (nephrin) antibody
    • NPHN antibody
    • NPHN_HUMAN antibody
    • NPHS 1 antibody
    • Nphs1 antibody
    • Renal glomerulus specific cell adhesion receptor antibody
    • Renal glomerulus-specific cell adhesion receptor antibody
    see all

Images

  • All lanes : Anti-Nephrin antibody (ab58968) at 1 µg/ml

    Lane 1 : mouse kidney tissue lysate
    Lane 2 : mouse kidney tissue lysate with blocking peptide

    Lysates/proteins at 15 µg per lane.

    Predicted band size: 138 kDa
    Observed band size: 100 kDa
    why is the actual band size different from the predicted?



    The difference in MW size could be due to modification to the protein or just aberrant migration.
  • ab58968, at a concentration of 1 µg/ml, staining Nephrin in mouse kidney tissue by Immunohistochemistry.
  • All lanes : Anti-Nephrin antibody (ab58968)

    Lane 1 : HEK293 cell lysate
    Lane 2 : Mouse kidney lysate

    Predicted band size: 138 kDa

  • Anti-Nephrin antibody (ab58968) at 0.5 µg/ml + Mouse kidney tissue lysate at 56 µg with Milk at 5 %

    Secondary
    HRP conjugated Goat anti-rabbit IgG monoclonal at 1/1000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 138 kDa
    Observed band size: 85 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds

    See Abreview

References

This product has been referenced in:
  • Zhang L  et al. PGC-1a ameliorates kidney fibrosis in mice with diabetic kidney disease through an antioxidative mechanism. Mol Med Rep 17:4490-4498 (2018). Read more (PubMed: 29344670) »
  • van de Lest NA  et al. Nephrin Loss Can Be Used to Predict Remission and Long-term Renal Outcome in Patients With Minimal Change Disease. Kidney Int Rep 3:168-177 (2018). Read more (PubMed: 29340328) »
See all 39 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Loading amount
56 µg
Gel Running Conditions
Reduced Denaturing (4-20% Tris Glycin)
Sample
Mouse Tissue lysate - whole (Kidney)
Specification
Kidney
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Aug 06 2013

Answer

Thank you for your email.
I am sorry to hear that these antibodies are not working as expected.
As for ab58968, we had replaced this antibody free of charge with ab85379. Since ab85379 was not tested in rat, sheep, and pig tissue, I had sent you a testing discount code. The code (xxx) is still valid until its expiration date of xxx.
Which species did you try ab85379 on?
As for ab50339, it has only been tested in WB, IHC-Fr, ICC/IF, but not paraffin embedded sections. It might work on paraffin sections, but we have no data to confirm this as it was not yet tested.
I'd be happy, though, to give you a testing discount code for trying it in IHC-P. Would you use it on the same species as ab85379?
I look forward to hear back and assist you further.
Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Answer

Thank you for your email. Please find below the tetsing discount code for ab85379. At the bottom of the email, I have listed the information regarding the free of charge replacement for ab15081 and ab58968.
DISCOUNT CODE: xxx
Expiration date: xxx
I am very pleased to hear you would like to accept our offer and test ab85379 in WB and rat/pig/sheep. This code will give you 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for WB and rat/pig/sheep and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.
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.

------------ FREE of charge REPLACEMENT ---------------------
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 these products 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 xxx with ab15081 lot GR66676-1 and ab85379. You should receive both items tomorrow.
To check the status of the order please contact our Customer Service team and reference this number.
Please note that the free of charge replacement vial for ab15081 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

Answer

Thank you for your reply and the additional information. This is very helpful to us for our internal
I have checked our stock, but we have, unfortunately, only a new lot for ab15081 - which I'd be happy to send you free of charge. For ab7671 and ab58968 we have currently no new lot in stock.
But for alpha 1 Sodium Potassium ATPase, we have several other antibodies that have been tested in IHC-P and rat: ab2872, ab2867 and ab74945.
For Nephrin, we have no other antibody that has been tested in rat, but we have 3 antibodies that were tested in IHC-P with human samples:ab85379, ab82113 and ab92887.
I could send you one of these antibodies free of charge together with a testing discount for trying them on rat samples. For more information on our testing discount program please see below.
Alternatively, a credit or refund can be issued.
For UNTESTED species and/or applications, we have established a testing discount program. Here is a brief description of how it works:
The testing discount program is for customers who like to use an antibody/protein on an untested species/application. You would purchase the antibody at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount for ONE antibody at the full price (100%) of the antibody you have tested. The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount.
Please let me know how you would like to proceed.
I look forward to hear back from you.

Read More

Answer

Thank you for contacting us.
I am sorry to hear that all 3 antibodies are not performing as expected.
Before proceeding with the free of charge replacements, could you please let me know the following experimental details:
1) What was the exact problem encountered (e.g. no signal, high background)?
2) What antibody dilution and incubation time was used?
3) What antigen retrieval step and blocking step were employed?
4) Is the detection system working well otherwise?
I look forward to hear back from you and resolve this problem.

Read More

Answer

Thank you for contacting us. I have checked with the originating lab and received the following reply:  "This has been brought to our attention a few times, and unfortunately, we do not have a good explanation.  It might be due to post-translational modification/cleavage event, but we have not read of any of that for Nephrin.  We suppose it may be detecting an alternative splicing product, but again, we have not seen anything like that mentioned in the literature. The sequence used as the antigen for this antibody does not possess any great homology to any other protein, and the antibody is obviously specific for this sequence. Sorry there is not a good explanation for this." Please be advised that the product will be covered under our garuantee. We guarantee all our products for tested species and applications as listed on the datasheet. The guarantee is valid for 6 months after a purchase. For more details about our Abpromise guarantee please check this link: https://www.abcam.com/abpromise. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information. 

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (kidney)
Loading amount
10 µg
Specification
kidney
Gel Running Conditions
Non-reduced Denaturing (7.5%)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Dr. Hongjie Wang

Verified customer

Submitted Mar 24 2011

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

Sign up