Recombinant Anti-CD98 antibody [EPR27111-83] (ab303510)

Flow cytometric analysis of mouse splenocytes cells labeling CD98 with AB303510 at 1/500 dilution (0.1 µg) (Right) compared with a Rabbit monoclonal IgG (ab172730) (Left) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells. Cells were stained with rabbit IgG or ab303510. Then stained with anti-CD4 conjugated to Alexa Fluor^® 647.

Flow cytometric analysis of NIH/3T3 (mouse embryonic fibroblast) cells labeling CD98 with AB303510 at 1/500 dilution (0.1 µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse kidney (fresh) tissue labeling CD98 with AB303510 at 1/500 dilution (0.988 µg/mL), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (2 µg/mL) (Green). Confocal image showing positive staining in proximal tubules of mouse kidney tissue. The nuclear counterstain was DAPI (Blue). The section was incubated with ab303510 for 60 mins at room temperature. The section was then mounted using Fluoromount^®. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbedat 1/1000 dilution (2 µg/mL).

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labeling CD98 with AB303510 at 1/50 dilution (9.88 µg/mL), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (2 µg/mL) (Green). Confocal image showing cytoplasmic and membranous staining in NIH/3T3 cell line. 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 dilution (2.5 µg/mL) (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 dilution (2 µg/mL).

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling CD98 with AB303510 at 1/1000 dilution (0.494 µg/mL), followed by a ready to use LeicaDS9800 (BOND^™ Polymer Refine Detection). Positive staining was observed on mouse kidney. The section was incubated with ab303510 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 dilution (BOND^™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used. 

Immunohistochemical analysis of paraffin-embedded mouse breast cancer tissue labeling CD98 with AB303510 at 1/1000 dilution (0.494 µg/mL), followed by a ready to use LeicaDS9800 (BOND^™ Polymer Refine Detection). Positive staining was observed on mouse breast cancer. The section was incubated with ab303510 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 dilution (BOND^™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used. 

Immunohistochemical analysis of paraffin-embedded mouse skeletal muscle tissue labeling CD98 with AB303510 at 1/1000 dilution (0.494 µg/mL), followed by a ready to use LeicaDS9800 (BOND^™ Polymer Refine Detection). Negative control: no staining is observed on mouse skeletal muscle. The section was incubated with ab303510 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 dilution (BOND^™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used. 

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

The slight variation of MW observed between the different samples is due to the different glycosylation.

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

The slight variation of MW observed between the different samples is due to the different glycosylation.

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

Low expression: skeletal muscle (human protein atlas database)

The molecular weight observed is consistent with what has been described in the literature (PMID: 18625289, 26611634).

This blot was developed using a high sensitivity ECL substrate.

The slight variation of MW observed between the different samples is due to the different glycosylation.

CD98 was immunoprecipitated from 0.35 mg 3T3-L1 (mouse embryonic fibroblast) whole cell lysate with ab303510 at 1/30 dilution (2 µg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab303510 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: 3T3-L1(mouse embryonic fibroblast) whole cell lysate 10 μg (Inset)

Lane 2: ab303510 IP in 3T3-L1whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab303510 in 3T3-L1 whole cell lysate

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

Exposure time: 180 seconds.

Observed M_W (kDa): 75-100