Anti-SLC1A5/ASCT2 antibody [CAL33]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SLC1A5/ASCT2 antibody [CAL33] (AB237704)

Immunofluorescent analysis of 100% methanol fixed Jurkat (human T cell leukemia T lymphocyte) cells labeling SLC1A5/ASCT2 with ab237704 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in Jurkat cells. The nuclear counter stain is DAPI (blue).
Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at a 1/200 dilution (red). The negative control is the secondary antibody only.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SLC1A5/ASCT2 antibody [CAL33] (AB237704)

Formalin-fixed, paraffin-embedded human colon carcinoma tissue stained for SLC1A5/ASCT2 using ab237704 at 0.5 μg/ml in immunohistochemical analysis.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SLC1A5/ASCT2 antibody [CAL33] (AB237704)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue labeling SLC1A5/ASCT2 with ab237704 at 1/1600 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Mainly membranous staining on the human lung carcinoma is observed. The section was incubated with ab237704 for 15 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP). Perform heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SLC1A5/ASCT2 antibody [CAL33] (AB237704)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue labeling SLC1A5/ASCT2 with ab237704 at 1/1600 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Mainly membranous staining on the human colon carcinoma is observed. The section was incubated with ab237704 for 15 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP). Perform heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SLC1A5/ASCT2 antibody [CAL33] (AB237704)

Immunofluorescent analysis of 100% methanol fixed HeLa (human cervix adenocarcinoma epithelial cell) cells labeling SLC1A5/ASCT2 with ab237704 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in HeLa cells. The nuclear counter stain is DAPI (blue).
Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at a 1/200 dilution (red). The negative control is the secondary antibody only.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-SLC1A5/ASCT2 antibody [CAL33] (AB237704)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized Jurkat (human T cell leukemia T lymphocyte) cell line labeling SLC1A5/ASCT2 with ab237704 at 1/40 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-SLC1A5/ASCT2 antibody [CAL33] (AB237704)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cell line labeling SLC1A5/ASCT2 with ab237704 at 1/400 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Collaborator

Immunocytochemistry/ Immunofluorescence - Anti-SLC1A5/ASCT2 antibody [CAL33] (AB237704)

HCT116 WT, SLC1A5 KO were labelled with a green dye. SLC1A5 KOOE cells were labelled with a green and a red dye. SLC1A5 expression was induced with doxycycline in KOOE cells. WT and KO cells were stained with ab237704 and with Alexa-fluor 594 coupled secondary antibody. KOOE cells were stained with : i) ab237704 and with Alexa-fluor 594 coupled secondary antibody; ii) anti-HA-Tag antibody and with Alexa-fluor 488 antibody. Acquisition of the green (HA-Tag in KOOE) and red (antibody staining in WT, KO and KOOE) channels was performed. Representative images of the green and red channel are shown. Antibodies used and tested dilutions are as follows : ab237704 at 1/50, 1/100 and 1/500.

This image was provided, with thanks, by RESOLUTE (re-solute.eu)

• IP

Lab

Immunoprecipitation - Anti-SLC1A5/ASCT2 antibody [CAL33] (AB237704)

SLC1A5/ASCT2 was immunoprecipitated from 0.35 mg Jurkat (human T cell leukemia T lymphocyte) whole cell lysate with ab237704 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab237704 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used at 1/5000 dilution.

Lane 1 : Jurkat whole cell lysate 10 μg (Input).
Lane 2 : ab237704 IP in Jurkat whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab237704 in Jurkat whole cell lysate.

Blocking/Dilution buffer : 5% NFDM/TBST.

ASCT2 (SLC1A5) can form dimers or trimers based on literature. (PMID : 23756778).

All lanes:

Immunoprecipitation - Anti-SLC1A5/ASCT2 antibody [CAL33] (ab237704) at 1/30 dilution

Predicted band size: 56 kDa

false

Exposure time: 3s

• IP

Lab

Immunoprecipitation - Anti-SLC1A5/ASCT2 antibody [CAL33] (AB237704)

SLC1A5/ASCT2 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab237704 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab237704 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used at 1/5000 dilution.

Lane 1 : HeLa whole cell lysate 10 μg (Input).
Lane 2 : ab237704 IP in HeLa whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab237704 in HeLa whole cell lysate.

Blocking/Dilution buffer : 5% NFDM/TBST.

ASCT2 (SLC1A5) can form dimers or trimers based on literature. (PMID : 23756778).

All lanes:

Immunoprecipitation - Anti-SLC1A5/ASCT2 antibody [CAL33] (ab237704) at 1/30 dilution

Predicted band size: 56 kDa

false

Exposure time: 1s

• WB

Lab

Western blot - Anti-SLC1A5/ASCT2 antibody [CAL33] (AB237704)

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

Exposure time :
Lane 1 : 80 seconds
Lane 2 : 40 seconds

An extra band around 50 kDa might be observed. We are not sure how to define it.

All lanes:

Western blot - Anti-SLC1A5/ASCT2 antibody [CAL33] (ab237704) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) whole lysate at 15 µg

Lane 2:

293T (Human embryonic kidney epithelial cell) whole lysate at 15 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 kDa

false