HA722088

Dishevelled 2 Recombinant Rabbit Monoclonal Antibody [PSH04-07]

cat.: HA722088

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P, FC
Clonality:	Monoclonal
Clone number:	PSH04-07

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 79 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within human Dishevelled 2 aa 550-700 / 736.
Positive control:	HeLa cell lysate, 293T cell lysate, MCF7 cell lysate, NCI-H1299 cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, HeLa, NIH/3T3, human colon tissue, mouse colon tissue, rat colon tissue.
Subcellular location:	Cell membrane, Cytoplasm, cytosol, Cytoplasmic vesicle, Nucleus.
Recommended Dilutions: WB IF-Cell IHC-P FC	1:2,000 1:100 1:50-1:200 1:1,000
Uniprot #:	SwissProt: O14641 Human \| Q60838 Mouse Entrez Gene: 303251 Rat
Alternative names:	Dishevelled 2 (homologous to Drosophila dsh) Dishevelled dsh homolog 2 dishevelled segment polarity protein 2 Dishevelled-2 Dishevelled2 DSH homolog 2 DVL 2 Dvl2 DVL2_HUMAN Segment polarity protein dishevelled homolog DVL 2 Segment polarity protein dishevelled homolog DVL-2 Segment polarity protein dishevelled homolog DVL2

Images

Fig1: Western blot analysis of Dishevelled 2 on different lysates with Rabbit anti-Dishevelled 2 antibody (HA722088) at 1/2,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: 293T cell lysate
Lane 3: MCF7 cell lysate
Lane 4: NCI-H1299 cell lysate
Lane 5: NIH/3T3 cell lysate
Lane 6: PC-12 cell lysate

Lysates/proteins at 30 µg/Lane.

Predicted band size: 79 kDa
Observed band size: 90 kDa

Exposure time: 1 minute;

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 (HA722088) 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.

Fig2: Immunocytochemistry analysis of HeLa cells labeling Dishevelled 2 with Rabbit anti-Dishevelled 2 antibody (HA722088) at 1/100 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-Dishevelled 2 antibody (HA722088) 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 (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.

	Fig3: Immunocytochemistry analysis of NIH/3T3 cells labeling Dishevelled 2 with Rabbit anti-Dishevelled 2 antibody (HA722088) at 1/100 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-Dishevelled 2 antibody (HA722088) 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 (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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-Dishevelled 2 antibody (HA722088) at 1/100 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 (HA722088) at 1/100 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-Dishevelled 2 antibody (HA722088) 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 (HA722088) 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.

Fig6: Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-Dishevelled 2 antibody (HA722088) 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 (HA722088) 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.

Fig7: Flow cytometric analysis of HeLa cells labeling Dishevelled 2.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA722088, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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