Anti-XCL1+XCL2 antibody [EPR26181-30] (BSA and Azide free)

• ICC

Supplier Data

Immunocytochemistry - Anti-XCL1+XCL2 antibody [EPR26181-30] (BSA and Azide free) (AB302523)

This data was developed using ab302522, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HEK-293T (human embryonic kidney epithelial cell) cells labelling XCL1+XCL2 with ab302522 at 1/100 (4.32 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 HEK-293T cells transfected with a human SCN2A expression vector containing a myc-tag. is observed. Myc-Tag Mouse mAb (Alexa Fluor® 647) was used to counterstain tubulin at 1/100 0.38ug/ml dilution (Red). 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.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-XCL1+XCL2 antibody [EPR26181-30] (BSA and Azide free) (AB302523)

This data was developed using ab302522, the same antibody clone in a different buffer formulation.Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Human peripheral blood mononuclear cell (PBMC) treated with 80nM TPA, 1.3uM Ionomycin, 10uM BFA and 2uM Monensin for 6h (Right) / Untreated control (Left) cells labelling XCL1+XCL2 with ab302522 at 1/500 dilution (0.1ug)/ Left and Right (Red) compared with a isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-XCL1+XCL2 antibody [EPR26181-30] (BSA and Azide free) (AB302523)

This data was developed using ab302522, the same antibody clone in a different buffer formulation.Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HuT-78 (human Sezary syndrome cutaneous T lymphocyte) treated with 80nM TPA and 3uM Ionomycin for 5h, then add 300ng/ml BFA for another 4h (Red) / Untreated control (Green) cells labelling XCL1+XCL2 with ab302522 at 1/500 dilution (0.1ug) (Red) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• ICC

Supplier Data

Immunocytochemistry - Anti-XCL1+XCL2 antibody [EPR26181-30] (BSA and Azide free) (AB302523)

This data was developed using ab302522, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HuT-78 (human Sezary syndrome cutaneous T lymphocyte) cells labelling XCL1+XCL2 with ab302522 at 1/1000 (0.432 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 increased cytoplasmic staining in HuT-78 cells treated with TPA (80 nM) and Ionomycin (3 uM) for 5 h, then add Brefeldin A (300ng/ml) for another 4 h is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). 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-XCL1+XCL2 antibody [EPR26181-30] (BSA and Azide free) (AB302523)

This data was developed using ab302522, the same antibody clone in a different buffer formulation.

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

All lanes:

Western blot - Anti-XCL1+XCL2 antibody [EPR26181-30] (<a href='/en-us/products/primary-antibodies/xcl1xcl2-antibody-epr26181-30-ab302522'>ab302522</a>) at 1/500 dilution

Lane 1:

Human XCL1 recombinant protein at 10 ng

Lane 2:

Human XCL2 recombinant protein at 10 ng

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: 12 kDa

Observed band size: 15 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-XCL1+XCL2 antibody [EPR26181-30] (BSA and Azide free) (AB302523)

This data was developed using ab302522, the same antibody clone in a different buffer formulation.

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

The expression of XCL1 are upregulated in response to TPA and Ionomycin treatment (PMID : 29474842, 32948687).

All lanes:

Western blot - Anti-XCL1+XCL2 antibody [EPR26181-30] (<a href='/en-us/products/primary-antibodies/xcl1xcl2-antibody-epr26181-30-ab302522'>ab302522</a>) at 1/500 dilution

Lane 1:

Untreated HuT-78 (human Sezary syndrome cutaneous T lymphocyte), whole cell lysate at 40 µg

Lane 2:

HuT-78 treated with 80 nM TPA and 3 uM Ionomycin for 5h, 300 ng/ml BFA was then added for additional 4h, whole cell lysate at 40 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: 12 kDa

Observed band size: 15 kDa

false

Exposure time: 1s