Wnt3a Recombinant Rabbit Monoclonal Antibody [PSH19-96]

Western blot analysis of Wnt3a on HCT 116 cell lysates with Rabbit anti-Wnt3a antibody (HA724116) at 1/5,000 dilution.

Lysates/proteins at 20 µg/Lane.

Predicted band size: 39 kDa
Observed band size: 39 kDa

Exposure time: 1 minute; ECL: K1802;

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 (HA724116) at 1/5,000 dilution was used in primary antibody dilution (K1803) 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 mouse E14.5 embryo tissue with Rabbit anti-Wnt3a antibody (HA724116) 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 (HA724116) 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.

Wnt3a was immunoprecipitated from 0.2 mg HCT 116 cell lysate with HA724116 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724116 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HCT 116 cell lysate (input)
Lane 2: HA724116 IP in HCT 116 cell lysate
Lane 3: Rabbit IgG instead of HA724116 in HCT 116 cell lysate

Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 3 minutes; ECL: K1802