Anti-CCL1 antibody [EPR28766-53] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CCL1 antibody [EPR28766-53] - BSA and Azide free (AB317572)

This data was developed using ab317571, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized U-937(human histiocytic lymphoma monocyte) cells labelling CCL1 with ab317571 at 1/50 (10.38 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing cytoplasmic staining in U-937 cells (shown in green) treated with 100 nM PMA for 72 hours, then together with human GM-CSF (1ug/ml) for another 24 hours, at last add Brefeldin A (5 µg/ml) for another 4 hours. The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CCL1 antibody [EPR28766-53] - BSA and Azide free (AB317572)

This data was developed using ab317571, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Jurkat (human T cell leukemia T lymphocyte from peripheral blood) cells labelling CCL1 with ab317571 at 1/50 (10.38 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing cytoplasmic staining in Jurkat cells (shown in green) treated with 10 ng/ml PMA, anti-CD28 (1ug/ml) and Brefeldin A (5 µg/ml) for 21 hours. The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• WB

Supplier Data

Western blot - Anti-CCL1 antibody [EPR28766-53] - BSA and Azide free (AB317572)

This data was developed using ab317571, the same antibody clone in a different buffer formulation.

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

The expression of CCL1 is upregulated in response to PMA, GM-CSF and BFA treatment.

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

All lanes:

Western blot - Anti-CCL1 antibody [EPR28766-53] (<a href='/en-us/products/primary-antibodies/ccl1-antibody-epr28766-53-ab317571'>ab317571</a>) at 1/1000 dilution

Lane 1:

U-937 (human histiocytic lymphoma monocyte) treated with 100nM PMA for 3 days whole cell lysate at 60 µg with 5% NFDM/TBST

Lane 2:

U-937 treated first with 100nM PMA for 3 days, then replaced with 50ng/ml GM-CSF for 24h, 5µg/ml BFA was then added for additional 4h at 60 µg with 5% NFDM/TBST

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: 15 kDa,124 kDa

false

Exposure time: 114s

• WB

Supplier Data

Western blot - Anti-CCL1 antibody [EPR28766-53] - BSA and Azide free (AB317572)

This data was developed using ab317571, the same antibody clone in a different buffer formulation.

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

The expression of CCL1 is upregulated in response to BFA, PMA and anti-CD28 treatment (PMID : 22413885).

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

All lanes:

Western blot - Anti-CCL1 antibody [EPR28766-53] (<a href='/en-us/products/primary-antibodies/ccl1-antibody-epr28766-53-ab317571'>ab317571</a>) at 1/1000 dilution

Lane 1:

Untreated Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 80 µg with 5% NFDM/TBST

Lane 2:

Jurkat treated with 300ng/ml BFA for 21h whole cell lysate at 80 µg with 5% NFDM/TBST

Lane 3:

Jurkat treated with 10ng/ml PMA and 1ug/ml anti-CD28 for 21h whole cell lysate at 80 µg with 5% NFDM/TBST

Lane 4:

Jurkat treated with 300ng/ml BFA, 10ng/ml PMA and 1ug/ml anti-CD28 for 21h whole cell lysate at 80 µg with 5% NFDM/TBST

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: 15 kDa,36 kDa

false

Exposure time: 37s