Anti-Ataxin 3 antibody [EPR24153-2] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Ataxin 3 antibody [EPR24153-2] - BSA and Azide free (AB284865)

This data was developed using ab259828, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized U-87 MG cells labelling Ataxin-3 with ab259828 at 1/50 (10.16 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing nuclear and cytoplasmic staining in U-87 MG cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Ataxin 3 antibody [EPR24153-2] - BSA and Azide free (AB284865)

This data was developed using ab259828, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling Ataxin-3 with ab259828 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, ab150081) at 1/2000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Ataxin 3 antibody [EPR24153-2] - BSA and Azide free (AB284865)

This data was developed using ab259828, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized U-87 MG (Human glioblastoma-astrocytoma epithelial cell) cells labelling Ataxin-3 with ab259828 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, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IP

Supplier Data

Immunoprecipitation - Anti-Ataxin 3 antibody [EPR24153-2] - BSA and Azide free (AB284865)

This data was developed using ab259828, the same antibody clone in a different buffer formulation.

Ataxin-3 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug with ab259828 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259828 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug

Lane 2 : ab259828 IP in HeLa whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab259828 in HeLa whole cell lysate

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

Exposure time : 15 seconds

Bands below 37kDa might be Ataxin-3 fragmemts(PMID : 30455355).

All lanes:

Immunoprecipitation - Anti-Ataxin 3 antibody [EPR24153-2] (<a href='/en-us/products/primary-antibodies/ataxin-3-antibody-epr24153-2-ab259828'>ab259828</a>)

Predicted band size: 42 kDa

false

• WB

Lab

Western blot - Anti-Ataxin 3 antibody [EPR24153-2] - BSA and Azide free (AB284865)

This data was developed using ab259828, the same antibody clone in a different buffer formulation.

Bands below 37kDa might be Ataxin-3 fragmemts(PMID : 30455355).

Exposure time : 70 seconds

All lanes:

Western blot - Anti-Ataxin 3 antibody [EPR24153-2] (<a href='/en-us/products/primary-antibodies/ataxin-3-antibody-epr24153-2-ab259828'>ab259828</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

Ataxin-3 knockdown HeLa whole cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 42 kDa

false

• WB

Lab

Western blot - Anti-Ataxin 3 antibody [EPR24153-2] - BSA and Azide free (AB284865)

This data was developed using ab259828, the same antibody clone in a different buffer formulation.

Bands below 37kDa might be Ataxin-3 fragmemts(PMID : 30455355).

Lanes 1-2 : 59 seconds

Lanes 3-4 : 3 minutes

All lanes:

Western blot - Anti-Ataxin 3 antibody [EPR24153-2] (<a href='/en-us/products/primary-antibodies/ataxin-3-antibody-epr24153-2-ab259828'>ab259828</a>) at 1/1000 dilution

Lane 1:

U-87 MG (human glioblastoma-astrocytoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

SH-SY5Y (human neuroblastoma epithelial cell), whole cell lysate at 20 µg

Lane 3:

Human brain tissue lysate at 20 µg

Lane 4:

Human heart tissue lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 42 kDa

false