Anti-VCAM1 antibody [EPR5038(2)] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-VCAM1 antibody [EPR5038(2)] - BSA and Azide free (AB249790)

This data was developed using ab174279, the same antibody clone in a different buffer formulation.

Immunoprecipitation. ab174279 at 1/1000 labeling VCAM1 immunoprecipitated from NIH3T3 cell lysate using ab174279 at 1/10.

All lanes:

Immunoprecipitation - Anti-VCAM1 antibody [EPR5038(2)] (<a href='/en-us/products/primary-antibodies/vcam1-antibody-epr50382-ab174279'>ab174279</a>)

Predicted band size: 81 kDa

Observed band size: 100 kDa

false

• WB

Lab

Western blot - Anti-VCAM1 antibody [EPR5038(2)] - BSA and Azide free (AB249790)

ab174279 was shown to react with VCAM1 in wild-type A549 cells in western blot. Loss of signal was observed when VCAM1 knockout sample was used. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab174279 overnight at 4 °C at a 1 in 1000 dilution. Blots were incubated with HRP conjugated Goat anti-Rabbit (H+L) secondary antibody at 1 in 5000 for 1 hour at room temperature before development with Optiblot ECL reagent (ab133456) and imaging.

All lanes:

Western blot - Anti-VCAM1 antibody [EPR5038(2)] (<a href='/en-us/products/primary-antibodies/vcam1-antibody-epr50382-ab174279'>ab174279</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 30 µg

Lane 2:

Wild-type A549 TNF-a treated (10 ng/mL, 16h) cell lysate at 30 µg

Lane 3:

VCAM1 knockout A549 cell lysate at 30 µg

Lanes 3 - 4:

Western blot - Human VCAM1 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-vcam1-knockout-a549-cell-line-ab273758'>ab273758</a>)

Lane 4:

VCAM1 knockout A549 TNF-a treated (10 ng/mL, 16h) cell lysate at 30 µg

Predicted band size: 81 kDa

Observed band size: 105 kDa

false

Exposure time: 20s

• WB

Supplier Data

Western blot - Anti-VCAM1 antibody [EPR5038(2)] - BSA and Azide free (AB249790)

This data was developed using ab174279, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-VCAM1 antibody [EPR5038(2)] (<a href='/en-us/products/primary-antibodies/vcam1-antibody-epr50382-ab174279'>ab174279</a>) at 1/1000 dilution

Lane 1:

Human fetal kidney lysate at 10 µg

Lane 2:

HuT-78 lysate at 10 µg

Lane 3:

NIH3T3 lysate at 10 µg

Predicted band size: 81 kDa

false

• WB

Unknown

Western blot - Anti-VCAM1 antibody [EPR5038(2)] - BSA and Azide free (AB249790)

This data was developed using ab174279, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-VCAM1 antibody [EPR5038(2)] (<a href='/en-us/products/primary-antibodies/vcam1-antibody-epr50382-ab174279'>ab174279</a>) at 1/1000 dilution

Lane 1:

Hut-78 (Human Sezary syndrome cutaneous T lymphocyte) whole cell lysate at 20 µg

Lane 2:

SK-OV-3 (Human ovarian cancer epithelial cell) whole cell lysate at 20 µg

Lane 3:

RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 4:

J774A.1 (Mouse reticulum cell sarcoma monocyte macrophage) whole cell lysate at 20 µg

Lane 5:

LADMAC (Mouse bone marrow monocyte macrophage) 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

Predicted band size: 81 kDa

Observed band size: 100 kDa

false

Exposure time: 15s

• WB

Unknown

Western blot - Anti-VCAM1 antibody [EPR5038(2)] - BSA and Azide free (AB249790)

This data was developed using ab174279, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-VCAM1 antibody [EPR5038(2)] (<a href='/en-us/products/primary-antibodies/vcam1-antibody-epr50382-ab174279'>ab174279</a>) at 1/1000 dilution

Lane 1:

HUVEC (Human umbilical vein endothelial cell) whole cell lysate at 20 µg

Lane 2:

HUVEC treated with 10 ng/ml TNF-α for 16 h, whole cell lysate at 20 µg

Lane 3:

bEND.3 (Mouse brain endothelioma) whole cell lysate at 20 µg

Lane 4:

bEND.3 treated with 10 μg/ml LPS for 24 h, 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

Predicted band size: 81 kDa

Observed band size: 100 kDa

false