Anti-CD98 antibody [EPR27110-42] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (AB307588)

This data was developed using ab307587, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human colon carcinom tissue labeling CD98 with ab307587 at 1/5000 (0.099 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human colon carcinoma (PMID : 29860836). The section was incubated with ab307587 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument.Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 10 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (AB307588)

This data was developed using ab307587, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling CD98 with ab307587 at 1/5000 (0.099 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on islet of human pancreas. The section was incubated with ab307587 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument.Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 10 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (AB307588)

This data was developed using ab307587, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling CD98 with ab307587 at 1/5000 (0.099 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human kidney. The section was incubated with ab307587 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 10 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (AB307588)

This data was developed using ab307587, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling CD98 with ab307587 at 1/100 (4.97 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 membrane and weak cytoplasmic staining in HeLa cell line.Low expression : SH-SY5YImage 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 2.5ug/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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (AB307588)

This data was developed using ab307587, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SLC3A2 (CD98) KO HeLa (SLC3A2 (CD98) knockout human cervix adenocarcinoma epithelial cell) (ab265708) cells labelling CD98 with ab307587 at 1/100 (4.97 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 membrane and weak cytoplasmic staining in parental HeLa 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 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 [EPR27110-42] - BSA and Azide free (AB307588)

This data was developed using ab307587, the same antibody clone in a different buffer formulation. Flow cytometric analysis of SH-SY5Y (human neuroblastoma epithelial cell, Left) / HeLa (human cervix adenocarcinoma epithelial cell, Right) cells labelling CD98 with ab307587 at 1/50 dilution (1ug) (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). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.Low expression : SH-SY5Y .

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (AB307588)

This data was developed using ab307587, the same antibody clone in a different buffer formulation. Flow cytometric analysis of Wild-type HeLa (human cervix adenocarcinoma epithelial cell, Right) / SLC3A2 (CD98) knockout HeLa (Left) cells labelling CD98 with ab307587 at 1/500 dilution (0.1ug) (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). Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.Positive staining on HeLa cells (ab255928), while no staining on CD98 knockout HeLa cells (ab265708).

• IP

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Immunoprecipitation - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (AB307588)

This data was developed using ab307587, the same antibody clone in a different buffer formulation. CD98 was immunoprecipitated from 0.35 mg HeLa whole cell lysate with ab307587 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307587 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate Lane 2 : ab307587 IP in HeLa whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab307587 in hela whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 1 second

All lanes:

Immunoprecipitation - Anti-CD98 antibody [EPR27110-42] (<a href='/en-us/products/primary-antibodies/cd98-antibody-epr27110-42-ab307587'>ab307587</a>) at 1/30 dilution

All lanes:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate

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: 1s

• WB

Supplier Data

Western blot - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (AB307588)

This data was developed using ab307587, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : SH-SY5Y Exposure time : 3 seconds

All lanes:

Western blot - Anti-CD98 antibody [EPR27110-42] (<a href='/en-us/products/primary-antibodies/cd98-antibody-epr27110-42-ab307587'>ab307587</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

SLC3A2 (CD98) knockout HeLa whole cell lysate at 20 µg

Lane 3:

HeLa whole cell lysate at 20 µg

Lane 4:

K562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 5:

SW620 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

SH-SY5Y (human neuroblastoma epithelial 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 1/100000 dilution

Observed band size: 70-130 kDa

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (AB307588)

This data was developed using ab307587, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : skeletal muscle (PMID : 10574970, PMID : 11557028). The lanes 2 and 3 were developed using a high sensitivity ECL substrate. Exposure time : Lane 1 : 6 seconds, lanes 2 and 3 : 48 seconds.

All lanes:

Western blot - Anti-CD98 antibody [EPR27110-42] (<a href='/en-us/products/primary-antibodies/cd98-antibody-epr27110-42-ab307587'>ab307587</a>) at 1/1000 dilution

Lane 1:

Human kidney tissue lysate at 20 µg

Lane 2:

Human breast tissue lysate at 20 µg

Lane 3:

Human 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 1/100000 dilution

Observed band size: 70-130 kDa

true

Exposure time: 6s