Anti-TIGAR antibody [EPR29002-30] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-TIGAR antibody [EPR29002-30] - BSA and Azide free (AB318981)

This data was developed using ab318980, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SW620 (human colorectal adenocarcinoma epithelial cell) cells labelling TIGAR with ab318980 at 1/50 (10.66 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 cytoplasmic and nuclear staining in SW620 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Low expression : K-562.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin 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.

• WB

Supplier Data

Western blot - Anti-TIGAR antibody [EPR29002-30] - BSA and Azide free (AB318981)

This data was developed using ab318980, the same antibody clone in a different buffer formulation.

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

Low expression : K-562, THP-1.

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

All lanes:

Western blot - Anti-TIGAR antibody [EPR29002-30] (<a href='/en-us/products/primary-antibodies/tigar-antibody-epr29002-30-ab318980'>ab318980</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

SW620 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

HCT-15 (human colon epithelial cell) whole cell lysate at 20 µg

Lane 4:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 5:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Western 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

Observed band size: 30 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-TIGAR antibody [EPR29002-30] - BSA and Azide free (AB318981)

This data was developed using ab318980, the same antibody clone in a different buffer formulation.

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

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

All lanes:

Western blot - Anti-TIGAR antibody [EPR29002-30] (<a href='/en-us/products/primary-antibodies/tigar-antibody-epr29002-30-ab318980'>ab318980</a>) at 1/1000 dilution

Lane 1:

Human brainstem tissue lysate at 20 µg

Lane 2:

Human hippocampus tissue lysate at 20 µg

Secondary

All lanes:

Western 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

Observed band size: 30 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-TIGAR antibody [EPR29002-30] - BSA and Azide free (AB318981)

This data was developed using ab318980, the same antibody clone in a different buffer formulation.

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

Doxorubicin treatment increased the TIGAR expression (PMID : 16839880).

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

All lanes:

Western blot - Anti-TIGAR antibody [EPR29002-30] (<a href='/en-us/products/primary-antibodies/tigar-antibody-epr29002-30-ab318980'>ab318980</a>) at 1/1000 dilution

Lane 1:

Untreated U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

U-2 OS treated with 0.2 μg/ml doxorubicin for 24 hours whole cell lysate at 20 µg

Lane 3:

Untreated MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

MCF7 treated with 0.5 μg/ml doxorubicin for 24 hours whole cell lysate at 20 µg

Secondary

All lanes:

Western 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

Observed band size: 30 kDa,36 kDa

false

Exposure time: 180s