Anti-Interferon beta antibody [RM1226]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Interferon beta antibody [RM1226] (AB322112)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized A549 (human lung carcinoma epithelial cell) cells labelling Interferon beta with ab322112 at 1/500 (0.946 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 A549 cells treated with 400 ng/ml poly(I : C) for 4 hours, then together with 300 ng/ml Brefeldin A (BFA) for another 20 hours (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). 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 (Magenta). 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-Interferon beta antibody [RM1226] (AB322112)

Interferon beta was immunoprecipitated from 0.35 mg A549 (human lung carcinoma epithelial cell) transfected with water instead of poly(I : C) for 24 hours, 300ng/ml BFA was added to cells 4 hours after the start of the transfection, whole cell lysate with ab322112 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab322112 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : A549 (human lung carcinoma epithelial cell) transfected with water instead of poly(I : C) for 24 hours, 300ng/ml BFA was added to cells 4 hours after the start of the transfection, whole cell lysate
Lane 2 : ab322112 IP in A549 (human lung carcinoma epithelial cell) transfected with water instead of poly(I : C) for 24 hours, 300ng/ml BFA was added to cells 4 hours after the start of the transfection, whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab322112 in A549 (human lung carcinoma epithelial cell) transfected with water instead of poly(I : C) for 24 hours, 300ng/ml BFA was added to cells 4 hours after the start of the transfection, whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-Interferon beta antibody [RM1226] (ab322112) at 1/30 dilution

All lanes:

A549 (human lung carcinoma epithelial cell) transfected with water instead of poly(I:C) for 24 hours, 300ng/ml BFA was added to cells 4 hours after the start of the transfection, 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: 111s

• IP

Supplier Data

Immunoprecipitation - Anti-Interferon beta antibody [RM1226] (AB322112)

Interferon beta was immunoprecipitated from 0.35 mg RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) treated with 50 ug/ml DMXAA (ab143018) and 300 ng/ml Brefeldin A (BFA) for 7 hours, whole cell lysate with ab322112 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab322112 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) treated with 50 ug/ml DMXAA (ab143018) and 300 ng/ml Brefeldin A (BFA) for 7 hours, whole cell lysate
Lane 2 : ab322112 IP in RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) treated with 50 ug/ml DMXAA (ab143018) and 300 ng/ml Brefeldin A (BFA) for 7 hours, whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab322112 in RAW 264.7 treated with 50 ug/ml DMXAA (ab143018) and 300 ng/ml Brefeldin A (BFA) for 7 hours, whole cell lysate

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

This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

All lanes:

Immunoprecipitation - Anti-Interferon beta antibody [RM1226] (ab322112) at 1/30 dilution

All lanes:

RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) treated with 50 ug/ml DMXAA (<a href='/en-us/products/unavailable/dmxaa-asa404-vegfr-inhibitor-ab143018'>ab143018</a>) and 300 ng/ml Brefeldin A (BFA) for 7 hours, 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: 41s

• WB

Supplier Data

Western blot - Anti-Interferon beta antibody [RM1226] (AB322112)

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

The expression profile observed is consistent with what has been described in the literature (PMID : 7352261, PMID : 23027866 and PMID : 16020513).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Interferon beta antibody [RM1226] (ab322112) at 1/1000 dilution

Lane 1:

A549 (human lung carcinoma epithelial cell) transfected with water instead of poly(I:C) for 24 hours, 300ng/ml BFA was added to cells 4 hours after the start of the transfection, whole cell lysate at 20 µg

Lane 2:

A549 transfected with 400ng/ml poly(I:C) for 24 hours, 300ng/ml BFA was added to cells 4 hours after the start of the transfection, 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 1/20000 dilution

Observed band size: 19 kDa,22 kDa,36 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-Interferon beta antibody [RM1226] (AB322112)

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

The expression profile observed is consistent with what has been described in the literature (PMID : 7352261, PMID : 23027866 and PMID : 16020513).

This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

The identity of the band around 75kDa is unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Interferon beta antibody [RM1226] (ab322112) at 1/1000 dilution

Lane 1:

Untreated RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 20 µg

Lane 2:

RAW 264.7 treated with 50 ug/ml DMXAA (<a href='/en-us/products/unavailable/dmxaa-asa404-vegfr-inhibitor-ab143018'>ab143018</a>) for 7 hours, then with 300 ng/ml Brefeldin A (BFA) added after 1 hour, whole cell lysate at 20 µg

Lane 3:

Untreated RAW 264.7 whole cell lysate at 20 µg

Lane 4:

RAW264.7 treated with 100 ng/ml lipopolysaccharide (LPS) for 6 hours, then with 300 ng/ml Brefeldin A (BFA) added after 3 hours, 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 1/20000 dilution

Observed band size: 24 kDa,26 kDa,36 kDa

true

Exposure time: 70s

• I-ELISA

Supplier Data

Indirect ELISA - Anti-Interferon beta antibody [RM1226] (AB322112)

Indirect ELISA analysis of ab322112 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution.

Antigen : Mouse IFN-β, Human IFN-β.

Antigen concentration : 1000 ng/ml