Anti-MAVS 抗体 [EPR26357-91] (BSA and Azide free) (ab290744)

This data was developed using ab290729, the same antibody clone in a different buffer formulation. 

Blocking / Diluting buffer and concentration:

5% NFDM/TBST

The molecular weight observed is consistent with that described in the literature (PMID: 16125763 and 30460894)
Lysates were prepared from fresh materials and used for Western blotting immediately to minimize protein degradation.

This data was developed using ab290729, the same antibody clone in a different buffer formulation. 

Immunohistochemical analysis of paraffin-embedded human skeletal muscle labeling MAVS with ab290729 at 1/500 dilution followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection). Positive cytoplasmic staining on Human skeletal muscle. The section was incubated with ab290729 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: PBS was used instead of primary antibody followed by ready to use secondary antibody  LeicaDS9800 (BOND™ Polymer Refine Detection).

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

This data was developed using ab290729, the same antibody clone in a different buffer formulation. 

Immunohistochemical analysis of paraffin-embedded human cardiac muscle labeling MAVS with ab290729 at 1/500 dilution  followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection). Positive cytoplasmic staining on human cardiac muscle is observed. The section was incubated with ab290729 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: PBS was used instead of primary antibody followed by ready to use secondary antibody  LeicaDS9800 (BOND™ Polymer Refine Detection).

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

This data was developed using ab290729, the same antibody clone in a different buffer formulation. 

Immunohistochemical analysis of paraffin-embedded human lung carcinoma labeling MAVS with ab290729 at 1/500 dilution followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection). Positive cytoplasmic staining on human lung carcinoma is observed. The section was incubated with ab290729 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: PBS was used instead of primary antibody followed by ready to use secondary antibody  LeicaDS9800 (BOND™ Polymer Refine Detection).

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

This data was developed using ab290729, the same antibody clone in a different buffer formulation. 

Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma labeling MAVS with ab290729 at 1/500 dilution followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection). Positive cytoplasmic staining on human ovarian carcinoma is observed. The section was incubated with ab290729 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: PBS was used instead of primary antibody followed by a ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection).

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

This data was developed using ab290729, the same antibody clone in a different buffer formulation. 

Immunofluorescent analysis of 4% paraformaldehyde 293T (human embryonic kidney epithelial) cells labeling MAVS with ab290729 at 1:50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed secondary antibody at 1/1000 dilution (Green). Confocal image showing mitochondrial staining on 293T cell line. ab33985 Anti-COX IV mouse monoclonal antibody - Mitochondrial Marker (1/1000 dilution) and ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (1/1000) was used to label mitochondria (RED) as a counterstain. DAPI was used as a nuclear counterstain (BLUE). 

The negative controls are as follows:
-ve control 1: Primary (ab290729) at 1/50 dilution, followed by secondary Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) at 1/1000 dilution.
-ve control 2:  Primary (ab33985) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150081) secondary at 1/1000 dilution.

This data was developed using ab290729, the same antibody clone in a different buffer formulation. 

Intracellular Flow Cytometry analysis of 293T (human embryonic kidney epithelial) cells labeling MAVS (red) with ab290729 at a 1/500 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) was used as the secondary antibody at a 1/2000 dilution. Black - isotype control: Rabbit monoclonal IgG (ab172730). Blue - unlabeled control: Cells without incubation with primary and secondary antibodies. 

This data was developed using ab290729, the same antibody clone in a different buffer formulation. 

MAVS was immunoprecipitated from 10 μg HeLa (human cervix adenocarcinoma epithelial cell) whole cell fresh lysate with ab290729 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab290729 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.

Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell fresh lysate 10 μg (Input).

Lane 2: ab290729 IP in HeLa whole cell fresh lysate. 

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab290744 in HeLa whole cell fresh lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.

Lysates were made fresh and used immediately to minimize protein degradation. The bands beneath the target band (75 kDa) are expected to be degradation products.

This data was developed using ab290729, the same antibody clone in a different buffer formulation. 

MAVS was immunoprecipitated from 10 μg 293T (human embryonic kidney epithelial cell) whole cell fresh lysate with ab290729 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab290729 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.

Lane 1: 293T (human embryonic kidney epithelial cell) whole cell fresh lysate 10 μg (Input).

Lane 2: ab290729 IP in 293T whole cell lysate. 

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab290744 in HeLa whole cell fresh lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.

The bands beneath the target band could be degradation or cleavage product, as demonstrated by the use of fresh lysate in the WB data.