Anti-CD10 antibody [EPR22867-118]

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-CD10 antibody [EPR22867-118] (AB256494)

Immunofluorescent analysis of 100% Methanol-fixed, Ramos (human Burkitt's lymphoma B lymphocyte) cells labelling CD10 with ab256494 at 1/500 dilution, followed by ab256494 anti- CD10 ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green).Confocal image showing membranous staining in Ramos cell line is observed. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue). Negative control : HT-29 （PMID : 19828468）

Secondary antibody only control : Secondary antibody is ab256494 anti- CD10 ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD10 antibody [EPR22867-118] (AB256494)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling CD10 with ab256494 at 1/500 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining on proximal convoluted tubules and glomerular epithelial cells of human kidney (PMID : 10705818) is observed. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD10 antibody [EPR22867-118] (AB256494)

Immunohistochemical analysis of paraffin-embedded Human placenta tissue labeling CD10 with ab256494 at 1/500 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous staining on syncytiotrophoblast layer of human placenta (PMID : 11092533) is observed. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD10 antibody [EPR22867-118] (AB256494)

Immunohistochemical analysis of paraffin-embedded Human diffuse large B-cell lymphoma labelling CD10 with ab256494 at 1/500 dilution, followed by a Goat Anti-Rabbit IgG H&L (HRP polymer) ready to use (ab214880). Positive staining on Human diffuse large B-cell lymphoma is observed. Counter stained with hematoxylin. Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a Goat Anti-Rabbit IgG H&L (HRP polymer) ready to use. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IP

Unknown

Immunoprecipitation - Anti-CD10 antibody [EPR22867-118] (AB256494)

CD10 was immunoprecipitated from 0.35 mg Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate with ab256494 at 1/30 (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab256494 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate 10ug

Lane 2 : ab256494 IP in Raji whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab256494 in Raji whole cell lysate

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

Exposure time : 3 min

All lanes:

Immunoprecipitation - Anti-CD10 antibody [EPR22867-118] (ab256494)

Predicted band size: 85 kDa

false

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD10 antibody [EPR22867-118] (AB256494)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling CD10 with ab256494 at 1/500 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous staining on renal tubules and of rat kidney (PMID : 10705818) is observed. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-CD10 antibody [EPR22867-118] (AB256494)

Immunofluorescent analysis of 100% Methanol-fixed WEHI-231 (mouse B cell lymphoma B lymphocyte) cells labelling CD10 with ab256494 at 1/500 dilution, followed by ab256494 anti- CD10 ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing staining in WEHI-231 cell line is observed. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab256494 anti- CD10 ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-CD10 antibody [EPR22867-118] (AB256494)

Immunohistochemical analysis of 4% PFA fixed 0.2% Triton X-100 permeabilized frozen Rat kidney tissue labeling CD10 with ab256494 at 1/100 (5.45 μg/ml) dilution followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution. The nuclear counterstain was DAPI (Blue). Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody was ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-CD10 antibody [EPR22867-118] (AB256494)

Immunofluorescent analysis of 100% Methanol-fixed 2.4G2 (rat B cell lymphoma B lymphocyte) cells labelling CD10 with ab256494 at 1/500 dilution, followed by ab256494 anti- CD10 ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing staining in 2.4G2 cell line is observed. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab256494 anti- CD10 ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD10 antibody [EPR22867-118] (AB256494)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling CD10 with ab256494 at 1/500 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous staining on renal tubules and of mouse kidney (PMID : 10705818) is observed. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-CD10 antibody [EPR22867-118] (AB256494)

Immunohistochemical analysis of 4% PFA fixed 0.2% Triton X-100 permeabilized frozen Mouse kidney tissue labeling CD10 with ab256494 at 1 : 100 (5.45 μg/ml) dilution followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution. The nuclear counterstain was DAPI (Blue). Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody was ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1 : 1000 (2 μg/ml) dilution.

• WB

Lab

Western blot - Anti-CD10 antibody [EPR22867-118] (AB256494)

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

The molecular weight observed is consistent with what has been described in the literature (PMID : 15286660)

Exposure time : Lane 1/3 : 8 seconds Lane 2 : 1 second Lane 4 : 3 seconds

All lanes:

Western blot - Anti-CD10 antibody [EPR22867-118] (ab256494) at 1/1000 dilution

Lane 1:

Rat lung tissue lysate at 10 µg

Lane 2:

Rat kidney tissue lysate at 10 µg

Lane 3:

Human tonsil tissue lysate at 10 µg

Lane 4:

Mouse lung tissue lysate 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

Predicted band size: 85 kDa

Observed band size: 100 kDa

false

• WB

Lab

Western blot - Anti-CD10 antibody [EPR22867-118] (AB256494)

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

ab256494 was shown to specifically react with CD10 in wild-type HAP1 cells as signal was lost in CD10 knockout cells. Wild-type and CD10 knockout samples were subjected to SDS-PAGE. ab256494 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging.

The molecular weight observed is consistent with what has been described in the literature (PMID : 15286660) Negative control : HT-29 (PMID : 19828468).

Exposure time : 59 seconds

All lanes:

Western blot - Anti-CD10 antibody [EPR22867-118] (ab256494) at 1/1000 dilution

Lane 1:

Wild type HAP1 whole cell lysate at 20 µg

Lane 2:

CD10 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

Raji (human Burkitts lymphoma B lymphocyte), whole cell lysate at 20 µg

Lane 4:

Ramos (human Burkitts lymphoma B lymphocyte), whole cell lysate at 20 µg

Lane 5:

HT-29 (human colorectal adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Secondary

Lanes 1 - 2:

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>)

Lanes 3 - 5:

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: 100 kDa

false