Recombinant
RabMAb

Recombinant Anti-Nephrin antibody [EPR20993] (ab216341)

Rabbit recombinant monoclonal Nephrin antibody [EPR20993]. Validated in WB, IP, IHC and tested in Mouse, Rat, Human. Cited in 2 publication(s).

Overview

  • Product name
    Anti-Nephrin antibody [EPR20993]
    See all Nephrin primary antibodies
  • Description
    Rabbit monoclonal [EPR20993] to Nephrin
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IP, IHC-P, WB, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Mouse Nephrin aa 300-1150. The exact sequence is proprietary.
    Database link: Q9QZS7

  • Positive control
    • WB: His-tagged mouse nephrin (aa1001-1250) recombinant protein; Mouse and rat kidney lysates. IHC-P: Human, mouse and rat kidney tissues. IHC-Fr: Mouse and rat kidney tissues. IP: Mouse kidney lysate.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    EPR20993
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab216341 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
IP 1/30.
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Use 1/2000 dilution for Mouse and Rat.

WB 1/1000. Detects a band of approximately 200 kDa (predicted molecular weight: 136 kDa).
IHC-Fr 1/500.

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

  • Anti-Nephrin antibody [EPR20993] (ab216341) at 1/10000 dilution + His-tagged mouse nephrin (aa1001-1250) recombinant protein at 0.01 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 136 kDa
    Observed band size: 38 kDa
    why is the actual band size different from the predicted?


    Exposure time: 15 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-Nephrin antibody [EPR20993] (ab216341) at 1/1000 dilution

    Lane 1 : Mouse kidney lysate
    Lane 2 : Mouse spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 136 kDa
    Observed band size: 200 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The MW is consistent with the literature (PMID 11733379).

  • Anti-Nephrin antibody [EPR20993] (ab216341) at 1/1000 dilution + Rat kidney lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Developed using the ECL technique.

    Predicted band size: 136 kDa
    Observed band size: 200 kDa why is the actual band size different from the predicted?


    Exposure time: 15 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Use high sensitivity ECL to obtain a strong signal.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Nephrin with ab216341 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous staining on human renal glomerulus (PMID: 18599604; PMID: 25367693). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling Nephrin with ab216341 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous staining on mouse renal glomerulus (PMID: 18599604; PMID: 25367693). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • Nephrin was immunoprecipitated from 0.35 mg of mouse kidney lysate with ab216341 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab216341 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution

    Lane 1: Mouse kidney lysate 10 µg (Input). 

    Lane 2: ab216341 IP in mouse kidney lysate. 

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab216341 in mouse kidney lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 30 seconds.

  • Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling Nephrin with ab216341 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous staining on rat renal glomerulus (PMID: 18599604; PMID: 25367693). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse kidney tissue labeling Nephrin with ab216341  at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Positive cytoplasmic staining in the glomerular podocytes (PMID: 26380915).

    The nuclear counter stain is DAPI (blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen rat kidney tissue labeling Nephrin with ab216341  at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Positive cytoplasmic staining in the glomerular podocytes (PMID: 26380915).

    The nuclear counter stain is DAPI (blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

References

This product has been referenced in:
  • Ling L  et al. Long noncoding RNA ENSRNOG00000037522 is involved in the podocyte epithelial-mesenchymal transition in diabetic rats. Int J Mol Med 41:2704-2714 (2018). Read more (PubMed: 29436579) »
  • He J  et al. The aldose reductase inhibitor epalrestat exerts nephritic protection on diabetic nephropathy in db/db mice through metabolic modulation. Acta Pharmacol Sin N/A:N/A (2018). Read more (PubMed: 29930278) »
See all 2 Publications for this product

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab216341.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
For licensing inquiries, please contact partnerships@abcam.com

Sign up