Anti-SLC7A5/LAT1 antibody [EPR26260-66] - BSA and Azide free

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-SLC7A5/LAT1 antibody [EPR26260-66] - BSA and Azide free (AB305252)

This data was developed using ab305251, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized LNCaP (human prostate carcinoma epithelial cell, Left) / HT-29 (human colorectal adenocarcinoma epithelial cell, Right) cells labelling SLC7A5/LAT1 with ab305251 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). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Negative control : LNCaP (PMID : 30241549).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-SLC7A5/LAT1 antibody [EPR26260-66] - BSA and Azide free (AB305252)

This data was developed using ab305251, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HT-29 (human colorectal adenocarcinoma epithelial cell) cells labelling SLC7A5/LAT1 with ab305251 at 1/100 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in HT-29 cell line. Negative control : LNCaP (PMID : 30241549). 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 (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.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SLC7A5/LAT1 antibody [EPR26260-66] - BSA and Azide free (AB305252)

• IP

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Immunoprecipitation - Anti-SLC7A5/LAT1 antibody [EPR26260-66] - BSA and Azide free (AB305252)

This data was developed using ab305251, the same antibody clone in a different buffer formulation. SLC7A5/LAT1 was immunoprecipitated from 0.35 mg HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate 10 μg with ab305251 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab305251. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate 10 μg Lane 2 : ab305251 IP in HT-29 whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab305251 in HT-29 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 3 seconds Samples are non-boiled as boiling may cause protein aggregates.

All lanes:

Immunoprecipitation - Anti-SLC7A5/LAT1 antibody [EPR26260-66] (<a href='/en-us/products/primary-antibodies/slc7a5-lat1-antibody-epr26260-66-ab305251'>ab305251</a>) at 1/30 dilution

All lanes:

HT-29 (human colorectal adenocarcinoma epithelial cell)

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

Observed band size: 40 kDa

false

Exposure time: 3s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SLC7A5/LAT1 antibody [EPR26260-66] - BSA and Azide free (AB305252)

• WB

Supplier Data

Western blot - Anti-SLC7A5/LAT1 antibody [EPR26260-66] - BSA and Azide free (AB305252)

This data was developed using ab305251, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : LNCaP (PMID : 30241549) Samples are non-boiled as boiling may cause protein aggregates (multi-pass membrane protein) Exposure time : 136 seconds

All lanes:

Western blot - Anti-SLC7A5/LAT1 antibody [EPR26260-66] (<a href='/en-us/products/primary-antibodies/slc7a5-lat1-antibody-epr26260-66-ab305251'>ab305251</a>) at 1/1000 dilution

Lane 1:

HT-29 (human colorectal adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

LNCaP (human prostate carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 3:

HT-1080 (human fibrosarcoma epithelial cell), whole cell lysate at 20 µg

Lane 4:

MCF7 (human breast adenocarcinoma 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: 40 kDa

false

Exposure time: 136s

• WB

Supplier Data

Western blot - Anti-SLC7A5/LAT1 antibody [EPR26260-66] - BSA and Azide free (AB305252)

This data was developed using ab305251, the same antibody clone in a different buffer formulation.

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

Exposure time : 1 second, left; 3 seconds, right.

All lanes:

Western blot - Anti-SLC7A5/LAT1 antibody [EPR26260-66] (<a href='/en-us/products/primary-antibodies/slc7a5-lat1-antibody-epr26260-66-ab305251'>ab305251</a>) at 1/1000 dilution

Lanes 1 and 3:

293T cells transfected with an empty vector containing a His tag whole cell lysate at 20 µg

Lanes 2 and 4:

293T cells transfected with a Human SLC7A5 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 1/20000 dilution

Predicted band size: 55 kDa

Observed band size: 37 kDa

false

Exposure time: 1s