Anti-Milk Fat Globule 1 antibody [EPR29114-57] - BSA and Azide free

• WB

Lab

Western blot - Anti-Milk Fat Globule 1 antibody [EPR29114-57] - BSA and Azide free (AB319153)

Performed under reducing conditions. In Western blot, ab319152 was shown to bind specifically to Milk Fat Globule 1. Target of interest was observed at 43 kDa in wild-type HEK-293 cell treated with 300 ng/ml Brefeldin A lysates (lane 2) with no signal observed at this size in MFGE8 knockout cell line (lanes 3-4) (lane 3, knockout cell line ab266132 / knockout cell lysate ab258514). This blot was developed using a high sensitivity ECL substrate.

All lanes:

Western blot - Anti-Milk Fat Globule 1 antibody [EPR29114-57] (<a href='/en-us/products/primary-antibodies/milk-fat-globule-1-antibody-epr29114-57-ab319152'>ab319152</a>) at 1/1000 dilution

Lane 1:

Untreated Wild-type HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

Wild-type HEK-293 treated with 300 ng/ml Brefeldin A (<a href='/en-us/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>) for 24 hours whole cell lysate at 20 µg

Lane 3:

Untreated MFGE8 knockout HEK-293 whole cell lysate at 20 µg

Lane 4:

MFGE8 knockout HEK-293 treated with 300 ng/ml Brefeldin A (<a href='/en-us/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>) 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: 43 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-Milk Fat Globule 1 antibody [EPR29114-57] - BSA and Azide free (AB319153)

This data was developed using ab319152, the same antibody clone in a different buffer formulation.

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

Low expression : Daudi, HL-60.

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

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

All lanes:

Western blot - Anti-Milk Fat Globule 1 antibody [EPR29114-57] (<a href='/en-us/products/primary-antibodies/milk-fat-globule-1-antibody-epr29114-57-ab319152'>ab319152</a>) at 1/1000 dilution

Lane 1:

Untreated A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

A549 treated with /ml Brefeldin A (<a href='/en-us/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>) for 24 hours whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

HEK-293 treated with 300 ng/ml Brefeldin A (<a href='/en-us/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>) for 24 hous whole cell lysate at 20 µg

Lane 5:

IMR-90 (human lung fibroblast) whole cell lysate at 20 µg

Lane 6:

MRC-5 (human lung fibroblast) whole cell lysate at 20 µg

Lane 7:

Daudi (human Burkitts lymphoma lymphoblast) whole cell lysate at 20 µg

Lane 8:

HL-60 (human acute promyelocytic leukemia promyeloblast) 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: 43 kDa,36 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Milk Fat Globule 1 antibody [EPR29114-57] - BSA and Azide free (AB319153)

This data was developed using ab319152, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer and concentration : 5% NFDM/TBST

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

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

All lanes:

Western blot - Anti-Milk Fat Globule 1 antibody [EPR29114-57] (<a href='/en-us/products/primary-antibodies/milk-fat-globule-1-antibody-epr29114-57-ab319152'>ab319152</a>) at 1/1000 dilution

Lane 1:

Untreated HEK-293 (human embryonic kidney epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HEK-293 transfected with scrambled siRNA control, then treated with 300 ng/ml Brefeldin A (<a href='/en-us/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>) for 24 hours whole cell lysate at 20 µg

Lane 3:

Untreated HEK-293 transfected with siRNA specifically targeting Milk Fat Globule 1 whole cell lysate at 20 µg

Lane 4:

HEK-293 transfected with siRNA specifically targeting Milk Fat Globule 1, then treated with 300/ml Brefeldin A (<a href='/en-us/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>) 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: 43 kDa,36 kDa

true

Exposure time: 180s