Anti-Interferon beta antibody [EPR23341-7] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Interferon beta antibody [EPR23341-7] - BSA and Azide free (AB275889)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab275880).

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized A549 (human lung carcinoma epithelial cell) cells labelling Interferon beta with ab275880 at 1/50 (10.14 µg/ml) dilution followed by a ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at a 1/1000 (2 µg) 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 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 (2.5µg/ml) dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at a 1/1000 (2 µg) dilution.

• IP

Unknown

Immunoprecipitation - Anti-Interferon beta antibody [EPR23341-7] - BSA and Azide free (AB275889)

This data was developed using ab275880, the same antibody clone in a different buffer formulation.

Interferon beta was immunoprecipitated from 0.35 mg A549 (human lung carcinoma epithelial cell) (transfected with 400ng/ml poly(I : C) for 4 hours, then 300ng/ml BFA was added for the last 20 hours) whole cell lysate with ab275880 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab275880 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : A549 (transfected with 400ng/ml poly(I : C) for 4 hours, then 300ng/ml BFA was added for the last 20 hours) whole cell lysate 10 ug

Lane 2 : ab275880 IP in A549 (transfected with 400ng/ml poly(I : C) for 4 hours, then 300ng/ml BFA was added for the last 20 hours) whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab275880 in A549 (transfected with 400ng/ml poly(I : C) for 4 hours, then 300ng/ml BFA was added for the last 20 hours) whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-Interferon beta antibody [EPR23341-7] (<a href='/en-us/products/primary-antibodies/interferon-beta-antibody-epr23341-7-ab275880'>ab275880</a>)

Predicted band size: 22 kDa

Observed band size: 19 kDa,22 kDa

false

• WB

Lab

Western blot - Anti-Interferon beta antibody [EPR23341-7] - BSA and Azide free (AB275889)

This data was developed using ab275880, the same antibody clone in a different buffer formulation.

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

The molecular weight observed is consistent with what has been described in the literature (PMID : 7352261).

Exposure time : 97 seconds.

All lanes:

Western blot - Anti-Interferon beta antibody [EPR23341-7] (<a href='/en-us/products/primary-antibodies/interferon-beta-antibody-epr23341-7-ab275880'>ab275880</a>) at 1/1000 dilution

Lane 1:

A549 (human lung carcinoma epithelial cell) 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

Lane 2:

A549 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

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: 22 kDa

Observed band size: 19 kDa,22 kDa

false

• WB

Lab

Western blot - Anti-Interferon beta antibody [EPR23341-7] - BSA and Azide free (AB275889)

This data was developed using ab275880, the same antibody clone in a different buffer formulation.

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

Exposure time : 3 seconds.

All lanes:

Western blot - Anti-Interferon beta antibody [EPR23341-7] (<a href='/en-us/products/primary-antibodies/interferon-beta-antibody-epr23341-7-ab275880'>ab275880</a>) at 1/1000 dilution

All lanes:

Recombinant Human IFN-beta protein, 10 ng

Secondary

All lanes:

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

Predicted band size: 22 kDa

Observed band size: 18 kDa

false