Recombinant Anti-Nephrin antibody [EPR20993] - BSA and Azide free (ab227806)
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 - BSA and Azide free
- Suitable for: IHC-Fr, IP, IHC-P, WB
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Nephrin antibody [EPR20993] - BSA and Azide free
See all Nephrin primary antibodies -
Description
Rabbit monoclonal [EPR20993] to Nephrin - BSA and Azide free -
Host species
Rabbit -
Specificity
Human species is recommended based on IHC-P results, we do not guarantee WB and IHC-Frozen for human.
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Tested applications
Suitable for: IHC-Fr, IP, IHC-P, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- IHC-P: Human kidney tissue.
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General notes
ab227806 is the carrier-free version of ab216341.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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
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
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20993 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab227806 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 |
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IHC-Fr |
Use at an assay dependent concentration.
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IP |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. 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. |
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WB |
Use at an assay dependent concentration. Detects a band of approximately 200 kDa (predicted molecular weight: 136 kDa).
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Notes |
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IHC-Fr
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. 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
Use at an assay dependent concentration. Detects a band of approximately 200 kDa (predicted molecular weight: 136 kDa). |
Target
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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
modificationsPhosphorylated 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
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Database links
- Entrez Gene: 4868 Human
- Entrez Gene: 54631 Mouse
- Entrez Gene: 64563 Rat
- Omim: 602716 Human
- SwissProt: O60500 Human
- SwissProt: Q9QZS7 Mouse
- SwissProt: Q9R044 Rat
- Unigene: 122186 Human
see all -
Alternative names
- CNF antibody
- Nephrin antibody
- Nephrosis 1 congenital Finnish type antibody
see all
Images
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216341).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Primary antibody incubation overnight at +4°C is recommended for human species
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216341).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216341).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216341).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216341).
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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 1/1000 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216341).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (5)
ab227806 has been referenced in 5 publications.
- Matoba K et al. ROCK2-induced metabolic rewiring in diabetic podocytopathy. Commun Biol 5:341 (2022). PubMed: 35396346
- Imeri F et al. Loss of sphingosine kinase 2 enhances Wilm's tumor suppressor gene 1 and nephrin expression in podocytes and protects from streptozotocin-induced podocytopathy and albuminuria in mice. Matrix Biol 98:32-48 (2021). PubMed: 34015468
- Solanki AK et al. Phosphorylation of slit diaphragm proteins NEPHRIN and NEPH1 upon binding of HGF promotes podocyte repair. J Biol Chem 297:101079 (2021). PubMed: 34391780
- Li Y et al. Anti-apoptosis mechanism of triptolide based on network pharmacology in focal segmental glomerulosclerosis rats. Biosci Rep 40:N/A (2020). PubMed: 32285909
- Wang J et al. Critical roles of PI3K/Akt/NF-?B survival axis in angiotensin II-induced podocyte injury. Mol Med Rep 20:5134-5144 (2019). PubMed: 31638199