Anti-Nephrin 抗体 [EPR20993] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nephrin antibody [EPR20993] - BSA and Azide free (AB227806)

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

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-Nephrin antibody [EPR20993] - BSA and Azide free (AB227806)

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).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Nephrin antibody [EPR20993] - BSA and Azide free (AB227806)

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).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nephrin antibody [EPR20993] - BSA and Azide free (AB227806)

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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nephrin antibody [EPR20993] - BSA and Azide free (AB227806)

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.

• IP

Supplier Data

Immunoprecipitation - Anti-Nephrin antibody [EPR20993] - BSA and Azide free (AB227806)

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.

All lanes:

Immunoprecipitation - Anti-Nephrin antibody [EPR20993] (<a href='/products/primary-antibodies/nephrin-antibody-epr20993-ab216341'>ab216341</a>)

Predicted band size: 134 kDa

false

• WB

Supplier Data

Western blot - Anti-Nephrin antibody [EPR20993] - BSA and Azide free (AB227806)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216341).

Blocking/Dilution buffer : 5% NFDM/TBST.

Use high sensitivity ECL to obtain a strong signal.

All lanes:

Western blot - Anti-Nephrin antibody [EPR20993] (<a href='/products/primary-antibodies/nephrin-antibody-epr20993-ab216341'>ab216341</a>) 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='/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] - BSA and Azide free (AB227806)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216341).

Blocking/Dilution buffer : 5% NFDM/TBST.

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

All lanes:

Western blot - Anti-Nephrin antibody [EPR20993] (<a href='/products/primary-antibodies/nephrin-antibody-epr20993-ab216341'>ab216341</a>) 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='/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] - BSA and Azide free (AB227806)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216341).

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Nephrin antibody [EPR20993] (<a href='/products/primary-antibodies/nephrin-antibody-epr20993-ab216341'>ab216341</a>) 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='/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