Sonic Hedgehog Protein/SHH(C-Product) Recombinant Rabbit Monoclonal Antibody [SC05-28]
cat.: ET1610-6
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse
Applications: WB, IF-Cell, IF-Tissue, IHC-P, FC
Clonality: Monoclonal
Clone number: SC05-28
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: 50 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human Sonic Hedgehog Protein aa 258-307 / 462.
Positive control: Mouse kidney tissue lysate, mouse brain tissue lysate, mouse liver tissue lysate, H22 cell lysate, Neuro-2a cell lysate, NIH/3T3 cell lysate, Neuro-2a, Hela, A549, 293, human liver cancer tissue, rat kidney tissue, rat liver tissue, human liver tissue, mouse liver tissue.
Subcellular location: Secreted, Cell membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
1:1,000-1:2,000
1:50-1:200
1:50-1:200
1:200-1:1,000
1:50-1:100
Uniprot #: SwissProt: Q15465 Human | Q62226 Mouse
Alternative names: HHG 1 HHG-1 HHG1 HLP 3 HLP3 Holoprosencephaly 3 HPE 3 HPE3 MCOPCB5 shh SHH_HUMAN SMMC I SMMCI Sonic Hedgehog (Drosophila) homolog sonic hedgehog homolog (Drosophila) Sonic hedgehog homolog Sonic hedgehog protein Sonic hedgehog protein C-product TPT TPTPS
Images
ET1610-6_1.jpg Fig1: Western blot analysis of Sonic Hedgehog Protein/SHH(C-Product) on different lysates with Rabbit anti-Sonic Hedgehog Protein/SHH(C-Product) antibody (ET1610-6) at 1/1,000 dilution.

Lane 1: Mouse kidney tissue lysate (30 µg/Lane)
Lane 2: Mouse brain tissue lysate (30 µg/Lane)
Lane 3: Mouse liver tissue lysate (30 µg/Lane)
Lane 4: H22 cell lysate (15 µg/Lane)
Lane 5: Neuro-2a cell lysate (15 µg/Lane)
Lane 6: NIH/3T3 cell lysate (15 µg/Lane)

Predicted band size: 50 kDa
Observed band size: 25/55 kDa

Exposure time: 30 seconds;

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 (ET1610-6) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.
ET1610-6_2.jpg Fig2: Immunocytochemistry analysis of Neuro-2a cells labeling Sonic Hedgehog Protein/SHH(C-Product) with Rabbit anti-Sonic Hedgehog Protein/SHH(C-Product) antibody (ET1610-6) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-Sonic Hedgehog Protein/SHH(C-Product) antibody (ET1610-6) 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.
ET1610-6_3.jpg Fig3: Western blot analysis of Sonic Hedgehog Protein on different lysates using anti-Sonic Hedgehog Protein antibody at 1/1,000 dilution.
Positive control:
Lane 1: Hela
Lane 2: HepG2
ET1610-6_4.jpg Fig4: ICC staining Sonic Hedgehog Protein in Hela cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
ET1610-6_5.jpg Fig5: ICC staining Sonic Hedgehog Protein in A549 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
ET1610-6_6.jpg Fig6: ICC staining Sonic Hedgehog Protein in 293 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
ET1610-6_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human liver cancer tissue using anti-Sonic Hedgehog Protein antibody. Counter stained with hematoxylin.
ET1610-6_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded rat kidney tissue using anti-Sonic Hedgehog Protein antibody. Counter stained with hematoxylin.
ET1610-6_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded rat liver tissue using anti-Sonic Hedgehog Protein antibody. Counter stained with hematoxylin.
ET1610-6_10.jpg Fig10: Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-Sonic Hedgehog Protein/SHH(C-Product) antibody (ET1610-6) 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 (ET1610-6) 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.
ET1610-6_11.jpg Fig11: Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-Sonic Hedgehog Protein/SHH(C-Product) antibody (ET1610-6) 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 (ET1610-6) 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.
ET1610-6_12.jpg Fig12: Flow cytometric analysis of Hela cells with Sonic Hedgehog Protein 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”.