Anti-IFIT1 antibody [EPR27276-63]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-IFIT1 antibody [EPR27276-63] (AB305301)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) treated with 10ng/ml human IFN alpha 1 for 16 hours (Red) / Untreated control (Green) cells labeling IFIT1 with ab305301 at 1/500 dilution (0.1ug) (Red) and Green (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IFIT1 antibody [EPR27276-63] (AB305301)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling IFIT1 with ab305301 at 1/2000 (0.261 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Cytoplasmic staining on human breast cancer (PMID : 23528130). The section was incubated with ab305301 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IFIT1 antibody [EPR27276-63] (AB305301)

Immunohistochemical analysis of paraffin-embedded HeLa cell pellet labeling IFIT1 with ab305301 at 1/2000 (0.261 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Cytoplasmic staining on HeLa cell pellet treated with IFN-alpha 1(10ng/ml) for 16 hours, no staining on HeLa cell pellet (untreated). The section was incubated with ab305301 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IFIT1 antibody [EPR27276-63] (AB305301)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue labeling IFIT1 with ab305301 at 1/2000 (0.261 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Cytoplasmic staining on human lung carcinoma (PMID : 35280763). The section was incubated with ab305301 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-IFIT1 antibody [EPR27276-63] (AB305301)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labeling IFIT1 with ab305301 at 1/50 (10.44 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 weak cytoplasmic staining in HeLa cell line, and the expression increased after treatment with IFN alpha 1 (10 ng/ml) for 16 hours. The 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 dilution (2.5 ug/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.

• IP

Supplier Data

Immunoprecipitation - Anti-IFIT1 antibody [EPR27276-63] (AB305301)

IFIT1 was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) treated with 10ng/ml IFN alpha 1 for 16h whole cell lysate 10 ug with ab305301 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab305301 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : HeLa treated with 10ng/ml IFN alpha 1 for 16h whole cell lysate 10 ug Lane 2 : ab305301 IP in HeLa treated with 10ng/ml IFN alpha 1 for 16h whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab305301 in HeLa whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 41 seconds.

All lanes:

Immunoprecipitation - Anti-IFIT1 antibody [EPR27276-63] (ab305301) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) treated with 10ng/ml IFN alpha 1 for 16h whole cell lysate 10 µg

Lane 2:

HeLa treated with 10ng/ml IFN alpha 1 for 16h 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

Observed band size: 56 kDa

false

Exposure time: 41s

• WB

Supplier Data

Western blot - Anti-IFIT1 antibody [EPR27276-63] (AB305301)

Blocking and diluting buffer and concentration : 5% NFDM/TBST   Exposure time : 180 seconds

All lanes:

Western blot - Anti-IFIT1 antibody [EPR27276-63] (ab305301) at 1/1000 dilution

Lane 1:

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

Lane 2:

HeLa treated with /ml IFN alpha 1 for 16h whole cell lysate 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: 56 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-IFIT1 antibody [EPR27276-63] (AB305301)

Blocking and diluting buffer and concentration : 5% NFDM/TBST   Exposure time : 26 seconds

All lanes:

Western blot - Anti-IFIT1 antibody [EPR27276-63] (ab305301) at 1/1000 dilution

Lane 1:

Human tonsil tissue lysate 40 µg

Lane 2:

Human testis tissue lysate 40 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 56 kDa

false

Exposure time: 26s