Anti-MX1 antibody [EPR19967] - BSA and Azide free

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-MX1 antibody [EPR19967] - BSA and Azide free (AB222492)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed Daudi (Human Burkitt's lymphoma cell line) treated with/without IFN-alpha (5000U/ml) for 48h cells labeling MX1 with ab207414 at 1/600 dilution (IFN alpha treated - red; untreated - green) compared with a 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 (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207414).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-MX1 antibody [EPR19967] - BSA and Azide free (AB222492)

This data was developed using ab207414, the same antibody clone in a different buffer formulation. Intracellular Flow Cytometry analysis of Daudi (Human Burkitt's lymphoma lymphoblast) treated with 20U/mL IFN alpha 1 for 24h cells labeling MX1 with purified ab207414 at 1/500 dilution (1μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cells without incubation with primary antibody and secondary antibody (Blue).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-MX1 antibody [EPR19967] - BSA and Azide free (AB222492)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Daudi (Human Burkitt's lymphoma cell line) cells labeling MX1 with ab207414 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing negative staining on Daudi cell line. Expression was induced and showed cytoplasmic staining after cells were treated with IFN alpha (5000 U/ml) for 48h. The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207414).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-MX1 antibody [EPR19967] - BSA and Azide free (AB222492)

Clone EPR19967 (ab222492) has been successfully conjugated by Abcam. This image was generated using Anti-MX1 antibody [EPR19967] (Alexa Fluor® 488). Please refer to ab237298 for protocol details.

ab237298 staining MX1 in Daudi cells +/- IFN alpha (5000 U/ml, 48h). The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab237298 at 1/50 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 594), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-MX1 antibody [EPR19967] - BSA and Azide free (AB222492)

Clone EPR19967 (ab222492) has been successfully conjugated by Abcam. This image was generated using Anti-MX1 antibody [EPR19967] (Alexa Fluor® 647). Please refer to ab237299 for protocol details.

ab237299 staining MX1 in Daudi cells +/- IFN alpha (5000 U/ml, 48h). The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab237299 at 1/50 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• IP

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Immunoprecipitation - Anti-MX1 antibody [EPR19967] - BSA and Azide free (AB222492)

MX1 was immunoprecipitated from 0.35 mg of Daudi (Human Burkitt's lymphoma cell line) treated with IFN alpha (5000U/ml) for 48h whole cell lysate with ab207414 at 1/40 dilution.

Western blot was performed from the immunoprecipitate using ab207414 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : Daudi treated with IFN alpha (5000U/ml) for 48h whole cell lysate 10μg (Input).

Lane 2 : ab207414 IP in Daudi treated with IFN alpha (5000U/ml) for 48h whole cell lysate.

Lane 3 : Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab207414 in Daudi treated with IFN alpha (5000U/ml) for 48h whole cell lysate.

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

Exposure time : 30 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207414).

All lanes:

Immunoprecipitation - Anti-MX1 antibody [EPR19967] - BSA and Azide free (ab222492)

Predicted band size: 76 kDa

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