Anti-TAP1 antibody [EPR26236-57] - BSA and Azide free

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-TAP1 antibody [EPR26236-57] - BSA and Azide free (AB314746)

This data was developed using ab314745, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HT-29 (human colorectal adenocarcinoma epithelial cell) cells labelling TAP1 with ab314745 at 1/50 (10.1 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 HT-29 cells treated with 100 ng/ml IFN gamma for 48 hours. 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 (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-TAP1 antibody [EPR26236-57] - BSA and Azide free (AB314746)

This data was developed using ab314745, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HT-29 (human colorectal adenocarcinoma epithelial cell) treated with 100ng/ml IFN gamma for 48h (Red) / Untreated HT-29 (Green) cells labelling TAP1 with ab314745 at 1/50 dilution (1 ug)/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). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IP

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Immunoprecipitation - Anti-TAP1 antibody [EPR26236-57] - BSA and Azide free (AB314746)

This data was developed using ab314745, the same antibody clone in a different buffer formulation. TAP1 was immunoprecipitated from 0.35 mg HT-29 (human colorectal adenocarcinoma epithelial cell) treated with 100ng/ml IFN gamma for 48h whole cell lysate with ab314745 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314745 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : HT-29 (human colorectal adenocarcinoma epithelial cell) treated with 100ng/ml IFN gamma for 48h whole cell lysate Lane 2 : ab314745 IP in HT-29 (human colorectal adenocarcinoma epithelial cell) treated with 100ng/ml IFN gamma for 48h whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab314745 in HT-29 treated with 100ng/ml IFN gamma for 48h whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-TAP1 antibody [EPR26236-57] (<a href='/en-us/products/primary-antibodies/tap1-antibody-epr26236-57-ab314745'>ab314745</a>) at 1/30 dilution

All lanes:

HT-29 (human colorectal adenocarcinoma epithelial cell) treated with 100ng/ml IFN gamma for 48h whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 5s

• WB

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Western blot - Anti-TAP1 antibody [EPR26236-57] - BSA and Azide free (AB314746)

This data was developed using ab314745, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST. The expression of TAP1 is upregulated in response to IFN gamma treatment (PMID : 12234057, PMID : 2475232). In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-TAP1 antibody [EPR26236-57] (<a href='/en-us/products/primary-antibodies/tap1-antibody-epr26236-57-ab314745'>ab314745</a>) at 1/1000 dilution

Lane 1:

Untreated HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HT-29 treated with /ml IFN gamma for 48h whole cell lysate at 20 µg

Lane 3:

Untreated HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

HeLa treated with 750U/ml IFN gamma for 48h whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Observed band size: 75 kDa

false

Exposure time: 7s

• Dot

Supplier Data

Dot Blot - Anti-TAP1 antibody [EPR26236-57] - BSA and Azide free (AB314746)

This data was developed using ab314745, the same antibody clone in a different buffer formulation. Dot blot analysis of TAP1 using ab314745 at 1 : 1000 (0.505 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution. Exposure time : 180 seconds. Blocking and diluting buffer and concentration : 5% NFDM/TBST. This antibody does not cross-react with human TAP2.