SAGE1 Recombinant Rabbit Monoclonal Antibody [PSH05-89] - BSA and Azide free

This data was developed using HA722487, the same antibody clone in a different buffer formulation.
Western blot analysis of SAGE1 on K-562 cell lysates with Rabbit anti-SAGE1 antibody (HA722487) at 1/1,000 dilution.

Lysates/proteins at 20 µg/Lane.

Predicted band size: 99 kDa
Observed band size: 120 kDa

Exposure time: 20 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 (HA722487) at 1/1,000 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.

This data was developed using HA722487, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of K-562 cells labeling SAGE1 with Rabbit anti-SAGE1 antibody (HA722487) at 1/500 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-SAGE1 antibody (HA722487) at 1/500 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.

This data was developed using HA722487, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-SAGE1 antibody (HA722487) 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 (HA722487) 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.

This data was developed using HA722487, the same antibody clone in a different buffer formulation.
SAGE1 was immunoprecipitated from 0.2 mg K-562 cell lysate with HA722487 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA722487 at 1/1,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs® VHH(HRP) (025-100-005) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: K-562 cell lysate (input)
Lane 2: HA722487 IP in K-562 cell lysate
Lane 3: Rabbit IgG instead of HA722487 in K-562 cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 3 minutes; ECL: K1801