Anti-Milk Fat Globule 1 antibody [EPR29114-57]

• WB

Lab

Western blot - Anti-Milk Fat Globule 1 antibody [EPR29114-57] (AB319152)

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] (ab319152) 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] (AB319152)

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] (ab319152) 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] (AB319152)

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] (ab319152) 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