Anti-CD10 antibody [EPR5904-110] - Low endotoxin, Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD10 antibody [EPR5904-110] - Low endotoxin, Azide free (AB222225)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling CD10 with ab208778 at 1/4000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Surface membrane staining was found in the glomerular epithelium and proximal tubular cells of the Human kidney. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is 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 (ab208778).

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-CD10 antibody [EPR5904-110] - Low endotoxin, Azide free (AB222225)

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling CD10 with ab208778 at 1/4000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Membrane staining was found in the subset cells of the Human breast cancer. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is 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 (ab208778).

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

• IP

Supplier Data

Immunoprecipitation - Anti-CD10 antibody [EPR5904-110] - Low endotoxin, Azide free (AB222225)

CD10 was immunoprecipitated from 1mg of LNCaP (Human prostate cancer cell line) whole cell lysate with ab208778 at 1/20 dilution.

Western blot was performed from the immunoprecipitate using ab208778 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : LNCaP whole cell lysate, 10μg (Input).

Lane 2 : ab208778 IP in LNCaP whole cell lysate.

Lane 3 : Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab208778 in LNCaP whole cell lysate.

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

Exposure time : 3 minutes.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab208778).

All lanes:

Immunoprecipitation - Anti-CD10 antibody [EPR5904-110] (<a href='/en-us/products/primary-antibodies/cd10-antibody-epr5904-110-ab208778'>ab208778</a>)

Predicted band size: 85 kDa

false

• WB

Supplier Data

Western blot - Anti-CD10 antibody [EPR5904-110] - Low endotoxin, Azide free (AB222225)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab208778).

Blocking/Dilution buffer : 5% NFDM/TBST.

Lane 1:

LNCaP (Human prostate cancer cell line) whole cell lysate at 10 µg

Lane 2:

Raji (Human Burkitt's lymphoma cell line) whole cell lysate at 10 µg

Lane 3:

Ramos (Human Burkitt's lymphoma cell line) whole cell 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: 85 kDa

Observed band size: 90-110 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-CD10 antibody [EPR5904-110] - Low endotoxin, Azide free (AB222225)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab208778).

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

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 1/10000 dilution

false

Exposure time: 5s

• WB

Unknown

Western blot - Anti-CD10 antibody [EPR5904-110] - Low endotoxin, Azide free (AB222225)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab208778).

Lane 1 : Wild type HAP1 whole cell lysate (20 μg)
Lane 2 : CD10 knockout HAP1 whole cell lysate (20 μg)
Lane 3 : RAJI whole cell lysate (20 μg)
Lanes 1 - 3 : Merged signal (red and green). Green - ab208778 observed at 100 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab208778 was shown to recognize CD10 when CD10 knockout samples were used, along with additional cross-reactive bands. Wild-type and CD10 knockout samples were subjected to SDS-PAGE. ab208778 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 100 dilution and 1/10000 dilution respectively. Blots were developed with 800CW Goat anti Rabbit and 680CW Goat anti Mouse secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CD10 antibody [EPR5904-110] - Low endotoxin, Azide free (ab222225)

Predicted band size: 85 kDa

false