Anti-CD98 antibody [EPR27111-83]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD98 antibody [EPR27111-83] (AB303510)

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.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD98 antibody [EPR27111-83] (AB303510)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling CD98 with ab303510 at 1/1000 (0.494 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining 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 (Bond™ Polymer Refine Detection).Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD98 antibody [EPR27111-83] (AB303510)

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.

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-CD98 antibody [EPR27111-83] (AB303510)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse kidney (fresh) tissue labeling CD98 with ab303510 at 1/500 (0.988 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (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 60mins 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). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CD98 antibody [EPR27111-83] (AB303510)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling CD98 with ab303510 at 1/50 (9.88 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 and membranous staining in NIH/3T3 cell line.Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (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 2 ug/ml dilution.

• Flow Cyt

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Flow Cytometry - Anti-CD98 antibody [EPR27111-83] (AB303510)

Flow cytometric analysis of / fixed / permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling CD98 with ab303510 at 1/500 dilution (0.1μg) (Red) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.

• Flow Cyt

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Flow Cytometry - Anti-CD98 antibody [EPR27111-83] (AB303510)

Flow cytometric analysis of / fixed / permeabilized Mouse splenocytes cells labelling CD98 with ab303510 at 1/500 dilution (0.1μg)/ Right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A 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.

• IP

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Immunoprecipitation - Anti-CD98 antibody [EPR27111-83] (AB303510)

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 MW (kDa) : 75-100

All lanes:

Immunoprecipitation - Anti-CD98 antibody [EPR27111-83] (ab303510) at 1/30 dilution

All lanes:

3T3-L1 (mouse embryonic fibroblast) whole cell lysate at 10 µg

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: 180s

• WB

Lab

Western blot - Anti-CD98 antibody [EPR27111-83] (AB303510)

This data was developed using ab303510, the same antibody clone in a different buffer formulation.  Blocking and dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Western blot - Anti-CD98 antibody [EPR27111-83] (ab303510) at 1/1000 dilution

Lane 1:

HEK-293T cells transfected with an empty vector containing a his tag, whole cell lysate at 20 µg

Lane 2:

HEK-293T cells transfected with a mouse CD98 expression vector containing a his tag, 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

Observed band size: 75-100 kDa

false

Exposure time: 10s

• WB

Lab

Western blot - Anti-CD98 antibody [EPR27111-83] (AB303510)

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 high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

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

All lanes:

Western blot - Anti-CD98 antibody [EPR27111-83] (ab303510) at 1/1000 dilution

Lane 1:

Mouse kidney tissue lysate at 20 µg

Lane 2:

Mouse placenta tissue lysate at 20 µg

Lane 3:

Mouse skeletal muscle tissue 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: 75-100 kDa

true

Exposure time: 26s

• WB

Lab

Western blot - Anti-CD98 antibody [EPR27111-83] (AB303510)

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.

All lanes:

Western blot - Anti-CD98 antibody [EPR27111-83] (ab303510) at 1/1000 dilution

Lane 1:

HC11 (mouse mammary gland epithelial cell), whole cell lysate at 20 µg

Lane 2:

ES-D3 (mouse blastocyst-derived embryonic stem cell), 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: 75-100 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-CD98 antibody [EPR27111-83] (AB303510)

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.

All lanes:

Western blot - Anti-CD98 antibody [EPR27111-83] (ab303510) at 1/1000 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 20 µg

Lane 2:

3T3-L1 (mouse embryonic fibroblast), 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: 75-100 kDa

false

Exposure time: 92s