Anti-MX1 antibody [EPR24485-19] - BSA and Azide free

• Flow Cyt

Lab

Flow Cytometry - Anti-MX1 antibody [EPR24485-19] - BSA and Azide free (AB284604)

This data was developed using ab284603, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Daudi (Human Burkitt's lymphoma lymphoblast) cells treated with 20 U/ml IFN alpha 1 for 24 hours (Red) or untreated (Green) labelling MX1 with ab284603 at 1/500 dilution (0.1ug) 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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MX1 antibody [EPR24485-19] - BSA and Azide free (AB284604)

This data was developed using ab284603, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labelling MX1 with ab284603 at 1/500 (1.09 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human breast cancer (PMID : 25838395). The section was incubated with ab284603 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 a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MX1 antibody [EPR24485-19] - BSA and Azide free (AB284604)

This data was developed using ab284603, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labelling MX1 with ab284603 at 1/500 (1.09 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human tonsil. The section was incubated with ab284603 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 a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval 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-MX1 antibody [EPR24485-19] - BSA and Azide free (AB284604)

This data was developed using ab284603, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labelling MX1 with ab284603 at 1/500 (1.09 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human spleen. The section was incubated with ab284603 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 a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-MX1 antibody [EPR24485-19] - BSA and Azide free (AB284604)

This data was developed using ab284603, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Daudi cells labelling MX1 with ab284603 at 1/250 (2.18 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 Daudi cell line, the signal increased after treatment with interferon alpha 1 (20 U/ml ) for 24 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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MX1 antibody [EPR24485-19] - BSA and Azide free (AB284604)

This data was developed using ab284603, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human colon cancer tissue labelling MX1 with ab284603 at 1/500 (1.09 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human colon cancer (PMID : 24771638). The section was incubated with ab284603 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 a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• WB

Lab

Western blot - Anti-MX1 antibody [EPR24485-19] - BSA and Azide free (AB284604)

This data was developed using ab284603, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration :

15 seconds

Exposure time :

All lanes:

Western blot - Anti-MX1 antibody [EPR24485-19] (<a href='/ko/products/primary-antibodies/mx1-antibody-epr24485-19-ab284603'>ab284603</a>) at 1/1000 dilution

Lane 1:

Untreated Daudi (human Burkitts lymphoma lymphoblast), whole cell lysate at 20 µg

Lane 2:

Daudi treated with 20U/ml IFN alpha for 24 hours at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 76 kDa

false

• WB

Lab

Western blot - Anti-MX1 antibody [EPR24485-19] - BSA and Azide free (AB284604)

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

Exposure time : 81 seconds

This data was developed using ab284603, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-MX1 antibody [EPR24485-19] (<a href='/ko/products/primary-antibodies/mx1-antibody-epr24485-19-ab284603'>ab284603</a>) at 1/1000 dilution

All lanes:

Human tonsil at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 76 kDa

false

• WB

Lab

Western blot - Anti-MX1 antibody [EPR24485-19] - BSA and Azide free (AB284604)

This data was developed using ab284603, the same antibody clone in a different buffer formulation.

Western blot : Rabbit Monoclonal[EPR24485-19] to MX1 ab284603 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 75 kDa. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-MX1 antibody [EPR24485-19] (<a href='/ko/products/primary-antibodies/mx1-antibody-epr24485-19-ab284603'>ab284603</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 (Vehicle Control for IFN alpha 0ng/mL for 24h) at 20 µg

Lane 2:

Wild-type A549 (IFN alpha 10ng/mL for 24h) at 20 µg

Lane 3:

MX1 knockout A549 (Vehicle Control for IFN alpha 0ng/mL for 24h) at 20 µg

Lane 4:

MX1 knockout A549 (IFN alpha 10ng/mL for 24h) at 20 µg

Lane 5:

Daudi (Vehicle Control for IFN alpha 0ng/mL for 24h) at 20 µg

Lane 6:

Daudi (IFN alpha 10ng/mL for 24h) at 20 µg

Lane 7:

LNCaP at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 76 kDa

Observed band size: 75 kDa,37 kDa

false

• WB

Lab

Western blot - Anti-MX1 antibody [EPR24485-19] - BSA and Azide free (AB284604)

This data was developed using ab284603, the same antibody clone in a different buffer formulation.

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

Exposure time : 15 seconds

All lanes:

Western blot - Anti-MX1 antibody [EPR24485-19] (<a href='/ko/products/primary-antibodies/mx1-antibody-epr24485-19-ab284603'>ab284603</a>) at 1/1000 dilution

Lane 1:

His-tagged human Interferon-induced GTP-binding protein Mx1 recombinant protein (aa37-366) at 0.01 µg

Lane 2:

His-tagged human Interferon-induced GTP-binding protein Mx2 recombinant protein (aa86-412) at 0.01 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 76 kDa

false