Antibodies, Proteins, Kits and Reagents for Life Science

291 484 ₸ Product size

100 µl 291 484 ₸

Order now and get it on Tuesday March 02, 2021

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

Product name

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

Rabbit monoclonal [EPR20993] to Nephrin
Host species

Rabbit

Tested Applications & Species

Application	Species
IHC-Fr	Mouse Rat
IHC-P	Mouse Rat Human
IP	Mouse
WB	Mouse Rat

See all applications and species data

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.

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

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)

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)

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)

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.
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)

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)

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)

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)

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)

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