Anti-Glutaminase antibody [EP7212] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glutaminase antibody [EP7212] - BSA and Azide free (AB214802)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human glioma tissue labelling Glutaminase with purified ab156876 at a dilution of 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab156876).

• WB

Lab

Western blot - Anti-Glutaminase antibody [EP7212] - BSA and Azide free (AB214802)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab156876).

Western blot : Anti-Glutaminase antibody [EP7212] ab156876 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. Three specific bands are detected by ab156876. We believe these correspond to isoform 1 (KGA), isoform 3 (GAC) and the mature mitochondrial subunits. Bands were observed at 68, 60 and 55 kDa in Wild-type MCF7 cell lysates with no signal observed at this size in GLS knockout MCF7 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Glutaminase antibody [EP7212] (<a href='/ko/products/primary-antibodies/glutaminase-antibody-ep7212-ab156876'>ab156876</a>) at 1/1000 dilution

Lane 1:

Wild-type MCF7 cell lysates at 20 µg

Lane 2:

Western blot - Human GLS knockout MCF7 cell line (<a href='/ko/products/cell-lines/human-gls-knockout-mcf7-cell-line-ab287444'>ab287444</a>) at 20 µg

Lane 3:

HeLa cell lysates at 20 µg

Lane 4:

OVCAR-3 cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 73 kDa

Observed band size: 68 kDa,60 kDa,55 kDa

false

• WB

Lab

Western blot - Anti-Glutaminase antibody [EP7212] - BSA and Azide free (AB214802)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab156876). Blocking and diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Glutaminase antibody [EP7212] (<a href='/ko/products/primary-antibodies/glutaminase-antibody-ep7212-ab156876'>ab156876</a>) at 1/1000 dilution

Lane 1:

HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 73 kDa

Observed band size: 65 kDa

true

Exposure time: 1s

• WB

Lab

Western blot - Anti-Glutaminase antibody [EP7212] - BSA and Azide free (AB214802)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab156876). Blocking and diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Glutaminase antibody [EP7212] (<a href='/ko/products/primary-antibodies/glutaminase-antibody-ep7212-ab156876'>ab156876</a>) at 1/1000 dilution

Lane 1:

Human kidney tissue lysate at 20 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 73 kDa

Observed band size: 65 kDa

true

Exposure time: 1s