Anti-Glutaredoxin 1 抗体

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glutaredoxin 1 antibody (AB45953)

IHC image of ab45953 staining in cervix formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab45953, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Glutaredoxin 1 antibody (AB45953)

ICC/IF image of ab45953 stained human HeLa cells. The cells were methanol fixed (5 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab45953, 1μg/ml) for 1h at room temperature. 1%BSA / 10% normal serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

• IP

Unknown

Immunoprecipitation - Anti-Glutaredoxin 1 antibody (AB45953)

Glutaredoxin 1 was immunoprecipitated using 0.5mg Jurkat whole cell extract, 5µg of Rabbit polyclonal to Glutaredoxin 1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Jurkat whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70^oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab45953.
Secondary : Clean-Blot IP Detection Reagent (HRP) at 1/500 dilution.
Band : 12kDa; Glutaredoxin 1.

All lanes:

Immunoprecipitation - Anti-Glutaredoxin 1 antibody (ab45953)

Predicted band size: 11 kDa

false

• WB

Project

Western blot - Anti-Glutaredoxin 1 antibody (AB45953)

All lanes:

Western blot - Anti-Glutaredoxin 1 antibody (ab45953) at 1/250 dilution

All lanes:

Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed at 1/3000 dilution

Predicted band size: 11 kDa

Observed band size: 12 kDa,45 kDa,70 kDa

false

• WB

Lab

Western blot - Anti-Glutaredoxin 1 antibody (AB45953)

Western blot : Rabbit Polyclonal to Glutaredoxin 1 ab45953 staining at 1/250 dilution, shown in black; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 10 kDa in Wild-type A549 cell lysates with no signal observed at this size in GLRX knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc 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 HRP (H+L) & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Glutaredoxin 1 antibody (ab45953) at 1/250 dilution

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

GLRX knockout A549 at 20 µg

Lane 3:

RKO at 20 µg

Lane 4:

A549 Nuclear at 20 µg

Secondary

Lanes 1 - 4:

Goat anti-Rabbit HRP (H+L) at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 12 kDa

Observed band size: 10 kDa

true

Exposure time: 90s

• WB

Unknown

Western blot - Anti-Glutaredoxin 1 antibody (AB45953)

All lanes:

Western blot - Anti-Glutaredoxin 1 antibody (ab45953) at 1/250 dilution

Lane 1:

Western blot - Recombinant Human Glutaredoxin 1 protein (Tag Free) (<a href='/products/proteins-peptides/recombinant-human-glutaredoxin-1-protein-ab86987'>ab86987</a>) at 0.01 µg

Lane 2:

Western blot - Recombinant Human Glutaredoxin 1 protein (Tag Free) (<a href='/products/proteins-peptides/recombinant-human-glutaredoxin-1-protein-ab86987'>ab86987</a>) at 0.001 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 11 kDa

true

Exposure time: 1min

• WB

Unknown

Western blot - Anti-Glutaredoxin 1 antibody (AB45953)

All lanes:

Western blot - Anti-Glutaredoxin 1 antibody (ab45953) at 1/250 dilution

Lane 1:

Western blot - Recombinant Human Glutaredoxin 1 protein (Tag Free) (<a href='/products/proteins-peptides/recombinant-human-glutaredoxin-1-protein-ab86987'>ab86987</a>) at 0.01 µg

Lane 2:

Western blot - Recombinant Human Glutaredoxin 1 protein (Tag Free) (<a href='/products/proteins-peptides/recombinant-human-glutaredoxin-1-protein-ab86987'>ab86987</a>) at 0.001 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 11 kDa

true

Exposure time: 1min