NSUN2 Recombinant Rabbit Monoclonal Antibody [PSH05-05]

Western blot analysis of NSUN2 on different lysates with Rabbit anti-NSUN2 antibody (HA722219) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: HepG2 cell lysate
Lane 3: Jurkat cell lysate
Lane 4: HEK-293 cell lysate
Lane 5: 293T cell lysate
Lane 6: A431 cell lysate
Lane 7: HCT 116 cell lysate
Lane 8: Caco-2 cell lysate
Lane 9: Neuro-2a cell lysate
Lane 10: COS-1 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 87 kDa
Observed band size: 100 kDa

Exposure time: 46 seconds; 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 (HA722219) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution were 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.

Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-NSUN2 antibody (HA722219) at 1/1,000 dilution and competitor's antibody at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA722219) at 1/1,000 dilution and competitor's antibody 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 human small intestine tissue with Rabbit anti-NSUN2 antibody (HA722219) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA722219) at 1/1,000 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.

Immunocytochemistry analysis of HeLa cells labeling NSUN2 with Rabbit anti-NSUN2 antibody (HA722219) at 1/250 dilution and competitor's antibody at 1/250 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-NSUN2 antibody (HA722219) at 1/250 dilution and competitor's antibody at 1/250 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 (M1305-2, 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.

NSUN2 was immunoprecipitated from 0.2 mg HeLa cell lysate with HA722219 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA722219 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HeLa cell lysate (input)
Lane 2: HA722219 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA722219 in HeLa cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 24 seconds; ECL: K1801