Anti-Interferon beta antibody [RM1226]

6 product images

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  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)(VeriBlot for IP Detection Reagent (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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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 (ab143018) and 300 ng/ml Brefeldin A (BFA) for 7 hours, whole cell lysate

  Secondary

  All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

  Exposure time: 41s

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  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)(VeriBlot for IP Detection Reagent (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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

  Exposure time: 111s

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  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 (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 (ab143018) 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) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Developed using the ECL technique.

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

  Exposure time: 70s

• 

  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 (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) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

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

  Exposure time: 15s

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  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

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  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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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).

  Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.