Anti-FAT/FAT1 antibody [EPR27361-28] - BSA and Azide free

• WB

Supplier Data

Western blot - Anti-FAT/FAT1 antibody [EPR27361-28] - BSA and Azide free (AB324787)

This data was developed using ab322343, the same antibody clone in a different buffer formulation.

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

Negative control : Raji (PMID : 22116550), K-562.

Full-length FAT1 is ~506kDa. The other bands are due to proteolytic cleavage in different FAT1 domains (PMID : 21680732).

The identity of the lower MW band at approximately 120 kDa is unknown.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-FAT/FAT1 antibody [EPR27361-28] (<a href='/en-us/products/primary-antibodies/fat-fat1-antibody-epr27361-28-ab322343'>ab322343</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 5:

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

Lane 6:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 7:

mIMCD3 (mouse inner medullary collecting duct epithelial cell) 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 1/20000 dilution

Observed band size: 506 kDa,124 kDa

false

Exposure time: 15s

• Dot

Supplier Data

Dot Blot - Anti-FAT/FAT1 antibody [EPR27361-28] - BSA and Azide free (AB324787)

This data was developed using ab322343, the same antibody clone in a different buffer formulation.

Dot blot analysis of FAT/FAT1 using ab322343 at 1/1000 (0.527 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane1 : Human FAT1 peptide
Lane2 : Human FAT3 peptide

Exposure time : 180 seconds.

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

This antibody does not cross-react with human FAT3.

All lanes:

Dot Blot - Anti-FAT/FAT1 antibody [EPR27361-28] (<a href='/en-us/products/primary-antibodies/fat-fat1-antibody-epr27361-28-ab322343'>ab322343</a>) at 1/1000 dilution

Lane 1:

Human FAT1 peptide

Lane 2:

Human FAT3 peptide

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

• WB

Supplier Data

Western blot - Anti-FAT/FAT1 antibody [EPR27361-28] - BSA and Azide free (AB324787)

This data was developed using ab322343, the same antibody clone in a different buffer formulation.

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

Low expression : spleen.

Full-length FAT1 is ~506kDa. The other bands are due to proteolytic cleavage in different FAT1 domains (PMID : 21680732).

The identity of the lower MW band at approximately 120 kDa is unknown.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-FAT/FAT1 antibody [EPR27361-28] (<a href='/en-us/products/primary-antibodies/fat-fat1-antibody-epr27361-28-ab322343'>ab322343</a>) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse spleen 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 1/20000 dilution

Observed band size: 506 kDa,124 kDa

false

Exposure time: 81s