MA3-082

antibody from Invitrogen Antibodies

Targeting: ATXN3 ATX3, JOS, MJD, SCA3

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA3-082

Provider

Invitrogen Antibodies

Product name

Ataxin 3 Monoclonal Antibody (2SCA-1H9)

Antibody type

Monoclonal

Antigen

Other

Description

MA3-082 detects Ataxin 3 from human, mouse and rat samples. MA3-082 has been successfully used in Western blot, immunofluorescence, and ELISA applications. MA3-082 has been successfully used in Western blot, immunofluorescence, and ELISA applications. The MA3-082 immunogen is the human ataxin-3 fragment from aa 112 - aa L249 as a fusion protein.

Reactivity

Human, Mouse, Rat

Host

Mouse

Isotype

IgG

Antibody clone number

2SCA-1H9

Vial size

50 µL

Concentration

Conc. Not Determined

Storage

-20° C, Avoid Freeze/Thaw Cycles

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Ataxin 3 was performed by loading 20 µg of the indicated whole cell lysates and 5 µL of PageRuler Plus Prestained Protein Ladder (Product # 26619) per well onto a 4-20% Tris-Glycine polyacrylamide gel (Product # WT4202BX10). Proteins were transferred to a nitrocellulose membrane using the G2 Blotter (Product # 62288), and blocked with 5% Milk in TBST for 1 hour at room temperature. Ataxin 3 was detected at 42 kDa using a Ataxin 3 mouse monoclonal antibody (Product # MA3-082) at a dilution of 1:500 in blocking buffer for 1 hour at room temperature on a rocking platform, followed by a Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177) at a dilution of 1:1000 for at least 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico substrat0e (Product # 34078) and the myECL Imager (Product # 62236).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Ataxin 3 was performed by loading 20 µg of the indicated whole cell lysates and 5 µL of PageRuler Plus Prestained Protein Ladder (Product # 26619) per well onto a 4-20% Tris-Glycine polyacrylamide gel (Product # WT4202BX10). Proteins were transferred to a nitrocellulose membrane using the G2 Blotter (Product # 62288), and blocked with 5% Milk in TBST for 1 hour at room temperature. Ataxin 3 was detected at 42 kDa using a Ataxin 3 mouse monoclonal antibody (Product # MA3-082) at a dilution of 1:500 in blocking buffer for 1 hour at room temperature on a rocking platform, followed by a Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177) at a dilution of 1:1000 for at least 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico substrat0e (Product # 34078) and the myECL Imager (Product # 62236).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Ataxin 3 was performed by loading 20 µg of HEK293T wildtype (Lane 1), HEK293T Ataxin 3 knockout (Lane 2) whole cell extracts. The blot was probed with Anti-Ataxin 3 Monoclonal Antibody (Product # MA3-082) (1:1000 dilution) and Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177) (1:4000 dilution). Loss of signal upon CRISPR mediated knockout (KO) confirms that antibody is specific to Ataxin 3.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of Ataxin 3 (green) in Caco-2 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and blocked with 3% BSA in PBS (Product # 37525) for 30 minutes at room temperature. Cells were stained with a Ataxin 3 mouse monoclonal antibody (Product # MA3-082) at a dilution of 1:100 in staining buffer for 1 hour at room temperature, and then incubated with a Goat anti-Mouse IgG Superclonal™ Secondary Antibody, Alexa Fluor 488 conjugate (Product # A28175) at a dilution of 1:1000 for 1 hour at room temperature. Nuclei (blue) were counterstained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.