Anti-AMID antibody [EPR29828-39] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-AMID antibody [EPR29828-39] - BSA and Azide free (AB326077)

This data was developed using ab324520, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed 0.1% TritonX-100 permeabilized DLD-1(human colorectal adenocarcinoma epithelial cell) cells labelling Ferroptosis suppressor protein 1 with ab324520 at 1/50 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in DLD-1 cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Low expression : T-47D
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 (1 ug/ml) dilution followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 (2 ug/ml) dilution (Magenta).

-ve control 1 : ab324520 at 1/50 dilution followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.
-ve control 2 : ab7291 at 1/1000 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution.

• WB

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Western blot - Anti-AMID antibody [EPR29828-39] - BSA and Azide free (AB326077)

This data was developed using ab324520, the same antibody clone in a different buffer formulation.

Milk as blocking agent for 1 hour. Concentration : 5%. Temperature : 25 °C.

Detection method : LICOR

The western blot shows validation of FSP1 knockout in Patu 8902 WT and FSP1 KO cell lines. For the knockout, two clones generated with sgRNA#1 and two clones generated with sgRNA#2 were used.

All lanes:

Western blot - Anti-Ferroptosis suppressor protein 1 antibody [EPR29828-39] (<a href='/ko/products/primary-antibodies/ferroptosis-suppressor-protein-1-antibody-epr29828-39-ab324520'>ab324520</a>) at 1/5000 dilution

Lane 1:

PATU 8902 (pancreas adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

FSP1 knockout (sgRNA &#35;1, clone 1) cell lysate at 20 µg

Lane 3:

FSP1 knockout (sgRNA &#35;1, clone 2) cell lysate at 20 µg

Lane 4:

FSP1 knockout (sgRNA &#35;2, clone 1) at 20 µg

Lane 5:

FSP1 knockout (sgRNA &#35;2, clone 2) cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG (IRDye® 800CW) at 1/5000 dilution

Observed band size: 40 kDa

false

Exposure time: 2min

This image is courtesy of an Abreview submitted by Mario Palma

• WB

Supplier Data

Western blot - Anti-AMID antibody [EPR29828-39] - BSA and Azide free (AB326077)

This data was developed using ab324520, the same antibody clone in a different buffer formulation.

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

Low expression : Jurkat (PMID : 11980907) T-47D JAR.

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

Exposure time : Lanes 1-6 : 70 seconds; Lane 7 : 180 seconds.

All lanes:

Western blot - Anti-Ferroptosis suppressor protein 1 antibody [EPR29828-39] (<a href='/ko/products/primary-antibodies/ferroptosis-suppressor-protein-1-antibody-epr29828-39-ab324520'>ab324520</a>) at 1/1000 dilution

Lane 1:

DLD-1 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

T-47D (human ductal breast epithelial tumor epithelial cell) whole cell lysate at 20 µg

Lane 3:

786-O (human kidney epithelial cell) whole cell lysate at 20 µg

Lane 4:

Jurkat (human t cell leukemia t lymphocyte from peripheral blood) whole cell lysate at 20 µg

Lane 5:

JAR (human placenta choriocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

A-172 (human brain glioblastoma cell ) whole cell lysate at 20 µg

Lane 7:

HepG2 (human hepatocellar carcinoma 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

Observed band size: 41 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-AMID antibody [EPR29828-39] - BSA and Azide free (AB326077)

This data was developed using ab324520, the same antibody clone in a different buffer formulation.

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

The identity of the band higher than 50 kDa is unknown.

All lanes:

Western blot - Anti-Ferroptosis suppressor protein 1 antibody [EPR29828-39] (<a href='/ko/products/primary-antibodies/ferroptosis-suppressor-protein-1-antibody-epr29828-39-ab324520'>ab324520</a>) at 1/1000 dilution

Lane 1:

Human lung tissue lysate at 20 µg

Lane 2:

Human spleen tissue lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 41 kDa

false

Exposure time: 180s