Anti-Nephrin antibody [EPR20993]

• IHC-P

Supplier Data

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

• IHC-Fr

Supplier Data

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.

• IHC-P

Supplier Data

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.

• IHC-P

Supplier Data

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.

• IHC-Fr

Unknown

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.

• IP

Supplier Data

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.

All lanes:

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

Predicted band size: 134 kDa

false

• WB

Supplier Data

Western blot - Anti-Nephrin antibody [EPR20993] (AB216341)

Blocking/Dilution buffer : 5% NFDM/TBST.

Use high sensitivity ECL to obtain a strong signal.

All lanes:

Western blot - Anti-Nephrin antibody [EPR20993] (ab216341) at 1/1000 dilution

All lanes:

Rat kidney lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 134 kDa

Observed band size: 200 kDa

true

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-Nephrin antibody [EPR20993] (AB216341)

Blocking/Dilution buffer : 5% NFDM/TBST.

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

All lanes:

Western blot - Anti-Nephrin antibody [EPR20993] (ab216341) at 1/1000 dilution

Lane 1:

Mouse kidney lysate at 10 µg

Lane 2:

Mouse spleen lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 134 kDa

Observed band size: 200 kDa

true

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-Nephrin antibody [EPR20993] (AB216341)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Nephrin antibody [EPR20993] (ab216341) at 1/10000 dilution

All lanes:

His-tagged mouse nephrin (aa1001-1250) recombinant protein at 0.01 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 134 kDa

Observed band size: 38 kDa

true

Exposure time: 15s

• WB

CiteAb

Western blot - Anti-Nephrin antibody [EPR20993] (AB216341)

Nephrin western blot using anti-Nephrin antibody [EPR20993] ab216341. Publication image and figure legend from Gong, Q., He, L. L., et al., 2019, Evid Based Complement Alternat Med, PubMed 31275420.

ab216341 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab216341 please see the product overview.

Fibrosis of kidney and expressions of the factors after the intravenous administration of B4. (a)–(f) Morphology of kidney (200×amplification). (a) normal; (b) model; (c) prednisolone (Predn.); (d) B4 (2.5 mg/kg); (e) B4 (1.25 mg/kg); (f) B4 (0.625 mg/kg); (g) the statistical analysis of the fibrosis in the kidney images by using grayscale scanning. (h)-(j) Protein expression of nephrin and podocin. The data were shown as mean ± SEM from 10 rats in each group. ##, compared with the normal control, p < 0.01. *, **, compared with model groups, p < 0.05 & p < 0.01. NS means no significance.

false