Anti-IFITM1 antibody [EPR22620-12]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IFITM1 antibody [EPR22620-12] (AB233545)

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling IFITM1 with ab233545 at 1/2000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on blood vessels of human cerebrum (PMID : 24603679). Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

The section was incubated with ab233545 for 10 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-IFITM1 antibody [EPR22620-12] (AB233545)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (human chronic myelogenous leukemia cell line from bone marrow) or A549 (human lung carcinoma cell line) cells labeling IFITM1 with ab233545 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in K-562 cells. Negative control : A549 (PMID : 25105503). The nuclear cpounter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

PBS only control : Use PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IFITM1 antibody [EPR22620-12] (AB233545)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling IFITM1 with ab233545 at 1/2000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on lymphocytes of human tonsil is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

The section was incubated with ab233545 for 10 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-IFITM1 antibody [EPR22620-12] (AB233545)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-fixed K562 (human chronic myelogenous leukemia cell line from bone marrow) (Right) or A549 (human lung carcinoma cell line) (Left) cells labeling IFITM1 with ab233545 at 1/500 (red) compared with a Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue).

Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.

Negative control : A549 (PMID : 25105503).

• IP

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Immunoprecipitation - Anti-IFITM1 antibody [EPR22620-12] (AB233545)

IFITM1 was immunoprecipitated from 0.35 mg of K562 (human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate with ab233545 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab233545 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.

Lane 1 : K562 whole cell lysate 10 μg (Input).
Lane 2 : ab233545 IP in K562 whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab233545 in K562 whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 3 seconds.

All lanes:

Immunoprecipitation - Anti-IFITM1 antibody [EPR22620-12] (ab233545)

Predicted band size: 13 kDa

Observed band size: 17 kDa

false

• WB

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Western blot - Anti-IFITM1 antibody [EPR22620-12] (AB233545)

Blocking and dilution buffer : 5% NFDM/TBST.

The molecular weight observed is consistent with what has been described in the literature (PMID : 25105503) IFN-α induce the expression of IFITM1(PMID : 27852071).

Negative control : A549 (PMID : 25105503).

All lanes:

Western blot - Anti-IFITM1 antibody [EPR22620-12] (ab233545) at 1/2000 dilution

Lane 1:

K562 (human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate at 20 µg

Lane 2:

A549 (human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

HeLa, treated with 10ng/ml hIFN-α for 16 hours, whole cell lysate at 20 µg

Lane 5:

TF-1 (human bone marrow erythroleukemia cell line) 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 1/100000 dilution

Predicted band size: 13 kDa

Observed band size: 17 kDa

false

Exposure time: 8s