SLC7A5 / LAT1 Recombinant Rabbit Monoclonal Antibody [PSH09-28] - BSA and Azide free

☑ Relative expression (RE)

Western blot analysis of SLC7A5 / LAT1 on different lysates with Rabbit anti-SLC7A5 / LAT1 antibody (HA751282) at 1/2,000 dilution.

Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: LNCaP cell lysate (negative) (20 µg/Lane)
Lane 3: Jurkat cell lysate (20 µg/Lane)
Lane 4: Mouse testis tissue lysate (40 µg/Lane)
Lane 5: Rat testis tissue lysate (40 µg/Lane)

Predicted band size: 55 kDa
Observed band size: 37 kDa

Exposure time: Lane 1-2: 1 minute; Lane 3-5: 3 minutes; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA751282) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

☑ Relative expression (RE)

Immunocytochemistry analysis of HT-29 (positive) and LNCaP (negative) labeling SLC7A5 / LAT1 with Rabbit anti-SLC7A5 / LAT1 antibody (HA751282) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-SLC7A5 / LAT1 antibody (HA751282) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-SLC7A5 / LAT1 antibody (HA751282) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA751282) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-SLC7A5 / LAT1 antibody (HA751282) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA751282) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

SLC7A5 / LAT1 was immunoprecipitated from 0.2 mg A549 cell lysate with HA751282 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751282 at 1/1,000 dilution. Mouse Anti-Rabbit IgG kappa light chain secondary antibody (M1208-2) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: A549 cell lysate (input)
Lane 2: HA751282 IP in A549 cell lysate
Lane 3: Rabbit IgG instead of HA751282 in A549 cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 59 seconds; ECL: K1801