Anti-UBE3A antibody [EPR23077-14] - BSA and Azide free

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-UBE3A antibody [EPR23077-14] - BSA and Azide free (AB272173)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol-permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling UBE3A with ab272168 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab272168).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-UBE3A antibody [EPR23077-14] - BSA and Azide free (AB272173)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells labelling UBE3A with ab272168 at 1/100 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear and weak cytoplasmic staining in HeLa cell line ab195889. Anti-alpha Tubulin antibody (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 ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab272168).

• I-ELISA

Unknown

Indirect ELISA - Anti-UBE3A antibody [EPR23077-14] - BSA and Azide free (AB272173)

indirect ELISA using ab272168 at varying antibody concentrations (1000-0 ng/ml) and Human UBE3A antigen at 1000 ng/ml. Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as a secondary antibody.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab272168).

• IP

Unknown

Immunoprecipitation - Anti-UBE3A antibody [EPR23077-14] - BSA and Azide free (AB272173)

UBE3A was immunoprecipitated from 0.35 mg K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate with ab272168 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab272168 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate 10 ug

Lane 2 : ab272168 IP in K-562 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab272168 in K-562 whole cell lysate

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

Exposure time : 15 seconds

A 37 kDa degraded band is observed.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab272168).

All lanes:

Immunoprecipitation - Anti-UBE3A antibody [EPR23077-14] (<a href='/en-us/products/primary-antibodies/ube3a-antibody-epr23077-14-ab272168'>ab272168</a>)

Predicted band size: 100 kDa

Observed band size: 100 kDa,37 kDa

false

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-UBE3A antibody [EPR23077-14] - BSA and Azide free (AB272173)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 cells labelling UBE3A with ab272168 at 1/100 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear and weak cytoplasmic staining in RAW 264.7 cell line. ab195889 Anti-alpha Tubulin antibody (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 ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab272168).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-UBE3A antibody [EPR23077-14] - BSA and Azide free (AB272173)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol-permeabilized RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) cells labelling UBE3A with ab272168 at 1/50 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab272168).

• IP

Unknown

Immunoprecipitation - Anti-UBE3A antibody [EPR23077-14] - BSA and Azide free (AB272173)

UBE3A was immunoprecipitated from 0.35 mg RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) (treated with 10 μM MG-132 for 4 hours) whole cell lysate with ab272168 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab272168 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) treated with 10 μM MG-132 for 4 hours, whole cell lysate 10 ug

Lane 2 : ab272168 IP in RAW 264.7 treated with 10 μM MG-132 for 4 hours, whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab272168 in RAW 264.7 treated with 10 μM MG-132 for 4 hours, whole cell lysate

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

Exposure time : 10 seconds

A 37 kDa degraded band is observed.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab272168).

All lanes:

Immunoprecipitation - Anti-UBE3A antibody [EPR23077-14] (<a href='/en-us/products/primary-antibodies/ube3a-antibody-epr23077-14-ab272168'>ab272168</a>)

Predicted band size: 100 kDa

Observed band size: 100 kDa,37 kDa

false