Dkk3 Recombinant Rabbit Monoclonal Antibody [JE35-98]
cat.: HA722396
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, IHC-Fr, IP
Clonality: Monoclonal
Clone number: JE35-98
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. 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: 38 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human Dkk3 aa 151-350 / 350.
Positive control: Mouse brain tissue lysate, mouse brain treated with deglycosylation tissue lysate, human brain tissue lysate, rat brain tissue lysate, HUVEC, human brain tissue, mouse brain tissue, rat brain tissue.
Subcellular location: Secreted.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  IHC-Fr
  IP
1:1,000
1:100
1:1,000
1:200
1-2μg/sample
Uniprot #: SwissProt: Q9UBP4 Human | Q9QUN9 Mouse
Entrez Gene: 171548 Rat
Alternative names: Dickkopf 3 Dickkopf 3 homolog (Xenopus laevis) Dickkopf 3 homolog Dickkopf homolog 3 Dickkopf related protein 3 Dickkopf related protein 3 Precursor Dickkopf WNT signaling pathway inhibitor 3 Dickkopf, Xenopus, homolog of, 3 Dickkopf-3 Dickkopf-related protein 3 Dickkopf3 DKK 3 Dkk-3 DKK3 DKK3_HUMAN hDkk 3 hDkk-3 hDkk3 Regulated in glioma REIC RIG RIG-like 5-6 RIG-like 7-1
Images
HA722396_1.jpg Fig1: Western blot analysis of Dkk3 on different lysates with Rabbit anti-Dkk3 antibody (HA722396) at 1/1,000 dilution.

Lane 1: Mouse brain tissue lysate (14 µg/Lane)
Lane 2: Mouse brain tissue lysate treated with deglycosylation (14 µg/Lane)
Lane 3: Human brain tissue lysate (20 µg/Lane)
Lane 4: Rat brain tissue lysate (20 µg/Lane)

Predicted band size: 38 kDa
Observed band size: 50-70 kDa

Exposure time: Lane 1-3: 10 seconds; Lane 4: 40 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 (HA722396) 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.
HA722396_2.jpg Fig2: Immunocytochemistry analysis of HUVEC cells labeling Dkk3 with Rabbit anti-Dkk3 antibody (HA722396) 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-Dkk3 antibody (HA722396) 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.
HA722396_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-Dkk3 antibody (HA722396) 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 (HA722396) 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.
HA722396_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-Dkk3 antibody (HA722396) 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 (HA722396) 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.
HA722396_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-Dkk3 antibody (HA722396) 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 (HA722396) 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.
HA722396_6.jpg Fig6: Immunofluorescence analysis of frozen mouse brain tissue with Rabbit anti-Dkk3 antibody (HA722396) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA722396, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
HA722396_7.jpg Fig7: Immunofluorescence analysis of frozen rat brain tissue with Rabbit anti-Dkk3 antibody (HA722396) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA722396, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
HA722396_8.jpg Fig8: Dkk3 was immunoprecipitated from 0.2 mg mouse brain tissue lysate with HA722396 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using HA722396 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: Mouse brain tissue lysate (input)
Lane 2: HA722396 IP in mouse brain tissue lysate
Lane 3: Rabbit IgG instead of HA722396 in mouse brain tissue lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 3 minutes; ECL: K1802
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.