Anti-Glutaminase antibody [EPR28716-72] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Glutaminase antibody [EPR28716-72] - BSA and Azide free (AB317033)

This data was developed using ab317032, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling Glutaminase with ab317032 at 1/500 (1.016 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 mitochondrial staining in NIH/3T3 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain at 1/200 (2.5ug/ml) dilution (Magenta). 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Glutaminase antibody [EPR28716-72] - BSA and Azide free (AB317033)

This data was developed using ab317032, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labelling Glutaminase with ab317032 at 1/500 (1.016 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 mitochondrial staining in Neuro-2a cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain at 1/200 (2.5ug/ml) dilution (Magenta). 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-Glutaminase antibody [EPR28716-72] - BSA and Azide free (AB317033)

This data was developed using ab317032, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling Glutaminase with ab317032 at 1/50 dilution (1 ug)/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).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• WB

Supplier Data

Western blot - Anti-Glutaminase antibody [EPR28716-72] - BSA and Azide free (AB317033)

This data was developed using ab317032, the same antibody clone in a different buffer formulation.

Negative control : liver (PMID : 19428809).

This antibody could detect 2 isoforms produced by alternative splicing.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lane 1-5 : 15 seconds, Lane 6 : 3 seconds, Lane 7 : 70 seconds

All lanes:

Western blot - Anti-Glutaminase antibody [EPR28716-72] (<a href='/en-us/products/primary-antibodies/glutaminase-antibody-epr28716-72-ab317032'>ab317032</a>) at 1/1000 dilution

Lane 1:

Mouse kidney tissue lysate at 50 µg

Lane 2:

Mouse liver tissue lysate at 50 µg

Lane 3:

Rat kidney tissue lysate at 50 µg

Lane 4:

Rat liver tissue lysate at 50 µg

Lane 5:

Human liver tissue lysate at 50 µg

Lane 6:

Human kidney tissue lysate at 20 µg

Lane 7:

Human lung tissue lysate at 50 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 66 kDa,73 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-Glutaminase antibody [EPR28716-72] - BSA and Azide free (AB317033)

This data was developed using ab317032, the same antibody clone in a different buffer formulation.

This antibody could detect 2 isoforms produced by alternative splicing.

Exposure time : Lane 1-4 : 10 seconds, Lane 5 : 15 seconds

All lanes:

Western blot - Anti-Glutaminase antibody [EPR28716-72] (<a href='/en-us/products/primary-antibodies/glutaminase-antibody-epr28716-72-ab317032'>ab317032</a>) at 1/1000 dilution

Lane 1:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 50 µg

Lane 2:

B16-F10 (Mouse skin melanoma), whole cell lysate at 50 µg

Lane 3:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 50 µg

Lane 4:

G-401(human rhabdoid tumor kidney epithelial cell) whole cell lysate at 50 µg

Lane 5:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 50 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 66 kDa,73 kDa

false

• Dot

Supplier Data

Dot Blot - Anti-Glutaminase antibody [EPR28716-72] - BSA and Azide free (AB317033)

This data was developed using ab317032, the same antibody clone in a different buffer formulation.

Dot blot analysis of Glutaminase using ab317032 at 1 : 1000 (0.508 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

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

This antibody does not cross-react with human/Mouse/Rat GLS2.

All lanes:

Dot Blot - Anti-Glutaminase antibody [EPR28716-72] (<a href='/en-us/products/primary-antibodies/glutaminase-antibody-epr28716-72-ab317032'>ab317032</a>) at 1/1000 dilution

Lane 1:

Mouse GLS1 peptide a

Lane 2:

Human GLS2 peptide b

Lane 3:

Mouse/Rat GLS2 peptide c

Lane 4:

Human GLS2 peptide d

Lane 5:

Mouse/Rat GLS2 peptide e

Secondary

All lanes:

Dot 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

false

Exposure time: 180s