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RecombinantRabMAb

Recombinant Anti-Nephrin antibody [EPR20993] (ab216341)

Reviews (1) Submit a question References (27)
Western blot - Anti-Nephrin antibody [EPR20993] (ab216341)
  • Western blot - Anti-Nephrin antibody [EPR20993] (ab216341)
  • Western blot - Anti-Nephrin antibody [EPR20993] (ab216341)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nephrin antibody [EPR20993] (ab216341)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nephrin antibody [EPR20993] (ab216341)
  • Immunoprecipitation - Anti-Nephrin antibody [EPR20993] (ab216341)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nephrin antibody [EPR20993] (ab216341)
  • Immunohistochemistry (Frozen sections) - Anti-Nephrin antibody [EPR20993] (ab216341)
  • Immunohistochemistry (Frozen sections) - Anti-Nephrin antibody [EPR20993] (ab216341)
  • Anti-Nephrin antibody [EPR20993] (ab216341)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20993] to Nephrin
  • Suitable for: IP, IHC-P, WB, IHC-Fr
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-Nephrin antibody [EPR20993]
    See all Nephrin primary antibodies

  • Description

    Rabbit monoclonal [EPR20993] to Nephrin

  • Host species

    Rabbit

  • Specificity

    Human species is recommended based on IHC-P results, we do not guarantee WB and IHC-Frozen for human.

  • Tested applications

    Suitable for: IP, IHC-P, WB, IHC-Frmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 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

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

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

    pH: 7.2
    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

The Abpromise guarantee

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.

Primary antibody incubation overnight at +4°C is recommended for human species.
WB
1/1000. Detects a band of approximately 200 kDa (predicted molecular weight: 136 kDa).
IHC-Fr
1/500.
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.

Primary antibody incubation overnight at +4°C is recommended for human species.
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

    see all

  • 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

  • Western blot - Anti-Nephrin antibody [EPR20993] (ab216341)
    Western blot - Anti-Nephrin antibody [EPR20993] (ab216341)
    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.

  • Western blot - Anti-Nephrin antibody [EPR20993] (ab216341)
    Western blot - Anti-Nephrin antibody [EPR20993] (ab216341)
    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).

  • Western blot - Anti-Nephrin antibody [EPR20993] (ab216341)
    Western blot - Anti-Nephrin antibody [EPR20993] (ab216341)
    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nephrin antibody [EPR20993] (ab216341)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nephrin antibody [EPR20993] (ab216341)

    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. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

    Primary antibody incubation overnight at +4°C is recommended for human species

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nephrin antibody [EPR20993] (ab216341)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nephrin antibody [EPR20993] (ab216341)

    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. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

  • Immunoprecipitation - Anti-Nephrin antibody [EPR20993] (ab216341)
    Immunoprecipitation - Anti-Nephrin antibody [EPR20993] (ab216341)

    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.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nephrin antibody [EPR20993] (ab216341)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nephrin antibody [EPR20993] (ab216341)

    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. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

  • Immunohistochemistry (Frozen sections) - Anti-Nephrin antibody [EPR20993] (ab216341)
    Immunohistochemistry (Frozen sections) - Anti-Nephrin antibody [EPR20993] (ab216341)

    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.

  • Immunohistochemistry (Frozen sections) - Anti-Nephrin antibody [EPR20993] (ab216341)
    Immunohistochemistry (Frozen sections) - Anti-Nephrin antibody [EPR20993] (ab216341)

    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.

  • Anti-Nephrin antibody [EPR20993] (ab216341)
    Anti-Nephrin antibody [EPR20993] (ab216341)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (27)

Publishing research using ab216341? Please let us know so that we can cite the reference in this datasheet.

ab216341 has been referenced in 27 publications.

  • Ding X  et al. MiR-21-5p in macrophage-derived extracellular vesicles affects podocyte pyroptosis in diabetic nephropathy by regulating A20. J Endocrinol Invest 44:1175-1184 (2021). PubMed: 32930981
  • Zang Y  et al. Astragaloside IV inhibits palmitic acid-induced apoptosis through regulation of calcium homeostasis in mice podocytes. Mol Biol Rep 48:1453-1464 (2021). PubMed: 33606151
  • Dong Z  et al. Inhibition of the Wnt/ß-catenin signaling pathway reduces autophagy levels in complement treated podocytes. Exp Ther Med 22:737 (2021). PubMed: 34055054
  • Sha S  et al. Mesenchymal stem cells-derived extracellular vesicles ameliorate Alzheimer's disease in rat models via the microRNA-29c-3p/BACE1 axis and the Wnt/ß-catenin pathway. Aging (Albany NY) 13:15285-15306 (2021). PubMed: 34086603
  • Xu X  et al. Adiponectin protects obesity-related glomerulopathy by inhibiting ROS/NF-?B/NLRP3 inflammation pathway. BMC Nephrol 22:218 (2021). PubMed: 34107901
View all Publications for this product

Customer reviews and Q&As

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IHC - Wholemount abreview for Anti-Nephrin antibody [EPR20993]

 Good
Abreviews
Application
IHC - Wholemount
Sample
Mouse Tissue (Heart)
Specification
Heart
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Verified customer

Submitted Dec 18 2020

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