Anti-UMOD antibody [EPR20071] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UMOD antibody [EPR20071] - BSA and Azide free (AB223540)

This IHC data was generated using the same anti-UMOD antibody clone [EPR20071] in a different buffer formulation (cat# ab207170).

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling UMOD with ab207170 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Strong cytoplasmic staining on distal convoluted is observed [PMID : 23988501].

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

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

• IP

Supplier Data

Immunoprecipitation - Anti-UMOD antibody [EPR20071] - BSA and Azide free (AB223540)

UMOD was immunoprecipitated from 0.35 mg of human kidney lysate with ab207170 at 1/30 dilution.

Western blot was performed from the immunoprecipitate using ab207170 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1 : Human kidney lysate 10μg (Input).

Lane 2 : ab207170 IP in human kidney lysate.

Lane 3 : Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab207170 in human kidney lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 0.5 second.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207170).

All lanes:

Immunoprecipitation - Anti-UMOD antibody [EPR20071] (<a href='/en-us/products/primary-antibodies/umod-antibody-epr20071-ab207170'>ab207170</a>)

Predicted band size: 70 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UMOD antibody [EPR20071] - BSA and Azide free (AB223540)

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling UMOD with ab207170 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Strong cytoplasmic staining on distal convoluted tubules of mouse kidney is observed [PMID : 23988501].

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207170).

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-UMOD antibody [EPR20071] - BSA and Azide free (AB223540)

Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling UMOD with ab207170 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Strong cytoplasmic staining on distal convoluted tubules of rat kidney is observed [PMID : 23988501].

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207170).

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UMOD antibody [EPR20071] - BSA and Azide free (AB223540)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207170).

Immunohistochemical analysis of paraffin-embedded (A) Kidney tissue from wild-type C57BL/6J mice and (B) Kidney tissue from Umod knockout mice labelling UMOD with ab207170 at 1/5000 (0.213 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) Kidney tissue from wild-type C57BL/6J mice, no staining on (B) Kidney tissue from UMOD knockout mice. The primary antibody was incubated for 10 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and Umod-KO homozygous mice (Strain ID : T014822).

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• IP

Supplier Data

Immunoprecipitation - Anti-UMOD antibody [EPR20071] - BSA and Azide free (AB223540)

UMOD was immunoprecipitated from 0.35 mg of mouse kidney lysate with ab207170 at 1/30 dilution.

Western blot was performed from the immunoprecipitate using ab207170 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1 : Human kidney lysate 10μg (Input).

Lane 2 : ab207170 IP in human kidney lysate.

Lane 3 : Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab207170 in human kidney lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 0.5 second.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207170).

All lanes:

Immunoprecipitation - Anti-UMOD antibody [EPR20071] (<a href='/en-us/products/primary-antibodies/umod-antibody-epr20071-ab207170'>ab207170</a>)

Predicted band size: 70 kDa

false

• WB

Supplier Data

Western blot - Anti-UMOD antibody [EPR20071] - BSA and Azide free (AB223540)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207170).

Blocking/Dilution buffer : 5% NFDM/TBST.

The MW observed is consistent with the literature (PMID : 18310901).

All lanes:

Western blot - Anti-UMOD antibody [EPR20071] (<a href='/en-us/products/primary-antibodies/umod-antibody-epr20071-ab207170'>ab207170</a>) at 1/10000 dilution

Lane 1:

Human urine at 10 µg

Lane 2:

Human fetal kidney lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/100000 dilution

Predicted band size: 70 kDa

Observed band size: 110 kDa

false

Exposure time: 5s

• WB

Supplier Data

Western blot - Anti-UMOD antibody [EPR20071] - BSA and Azide free (AB223540)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207170).

Blocking/Dilution buffer : 5% NFDM/TBST.

The MW observed is consistent with the literature (PMID : 18310901).

All lanes:

Western blot - Anti-UMOD antibody [EPR20071] (<a href='/en-us/products/primary-antibodies/umod-antibody-epr20071-ab207170'>ab207170</a>) at 1/1000 dilution

Lane 1:

Mouse kidney lysate at 10 µg

Lane 2:

Rat kidney lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 70 kDa

Observed band size: 110 kDa

false

Exposure time: 1s

• WB

Lab

Western blot - Anti-UMOD antibody [EPR20071] - BSA and Azide free (AB223540)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207170).

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and Umod-KO homozygous mice (Strain ID : T014822).

All lanes:

Western blot - Anti-UMOD antibody [EPR20071] (<a href='/en-us/products/primary-antibodies/umod-antibody-epr20071-ab207170'>ab207170</a>) at 1/1000 dilution

Lane 1:

Wild-type mouse kidney tissue lysate (male) at 20 µg

Lane 2:

Wild-type mouse kidney tissue lysate (female) at 20 µg

Lane 3:

Umod knockout mouse kidney tissue lysate (male case1) at 20 µg

Lane 4:

Umod knockout mouse kidney tissue lysate (male case2) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Observed band size: 120 kDa

false

Exposure time: 4s