Wnt5a Recombinant Rabbit Monoclonal Antibody [JU30-30]
cat.: ET1706-33
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IF-Cell, IHC-P, FC
Clonality: Monoclonal
Clone number: JU30-30
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 42 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human Wnt5a aa 331-380 / 380.
Positive control: Hela cell lysates, PC-3M, human lung cancer tissue, SH-SY5Y.
Subcellular location: Secreted, extracellular space, extracellular matrix.
Recommended Dilutions:
  WB
  IHC-P
  IF-Cell
  FC

1:500-1:2,000
1:1,000
1:50
1:50-1:100
Uniprot #: SwissProt: P41221 Human
Alternative names: hWNT 5A hWNT5A Protein Wnt 5a Protein Wnt-5a Protein Wnt5a Wingless type MMTV integration site family member 5A Wnt 5a WNT 5A protein WNT 5A protein precursor WNT5A WNT5A protein precursor WNT5A_HUMAN
Images
ET1706-33_1.jpg Fig1: Western blot analysis of Wnt5a on Hela cell lysates with Rabbit anti-Wnt5a antibody (ET1706-33) at 1/500 dilution.

Lysates/proteins at 10 µg/Lane.

Predicted band size: 42 kDa
Observed band size: 42 kDa

Exposure time: 2 minutes;

10% 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 (ET1706-33) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
ET1706-33_2.jpg Fig2: Immunocytochemistry analysis of PC-3M cells labeling Wnt5a with Rabbit anti-Wnt5a antibody (ET1706-33) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Wnt5a antibody (ET1706-33) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
ET1706-33_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human lung cancer tissue with Rabbit anti-Wnt5a antibody (ET1706-33) at 1/1,000 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 (ET1706-33) 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.
ET1706-33_4.jpg Fig4: Flow cytometric analysis of SH-SY5Y cells with Wnt5a antibody at 1/50 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.