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Sonic Hedgehog Protein/SHH(C-Product) Recombinant Rabbit Monoclonal Antibody [SC05-28]

Usd: 385

Specification

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Catalog# ET1610-6

Sonic Hedgehog Protein/SHH(C-Product) Recombinant Rabbit Monoclonal Antibody [SC05-28]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Sonic Hedgehog Protein/SHH(C-Product) Recombinant Rabbit Monoclonal Antibody [SC05-28]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human Sonic Hedgehog Protein aa 258-307 / 462.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P, FC

Molecular Weight

Predicted band size: 50 kDa

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, Rat heart tissue lysate, Rat liver tissue lysate, Rat brain tissue lysate, Rat lung tissue lysate, Neuro-2a, Hela, human liver tissue, mouse liver tissue, rat kidney tissue, rat liver tissue.

Conjugation

unconjugated

Clone Number

SC05-28

RRID

AB_3069944

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:2,000

  • IF-Cell

  • 1:50-1:200

  • IF-Tissue

  • 1:50-1:200

  • IHC-P

  • 1:200-1:1,000

  • FC

  • 1:50-1:100

Target

Function

Sonic hedgehog protein (SHH) is a major signaling molecule of embryonic development in humans and animals, encoded by the SHH gene. This signaling molecule is key in regulating embryonic morphogenesis in all animals. SHH controls organogenesis and the organization of the central nervous system, limbs, digits and many other parts of the body. Sonic hedgehog is a morphogen that patterns the developing embryo using a concentration gradient characterized by the French flag model. This model has a non-uniform distribution of SHH molecules which governs different cell fates according to concentration. Mutations in this gene can cause holoprosencephaly, a failure of splitting in the cerebral hemispheres, as demonstrated in an experiment using SHH knock-out mice in which the forebrain midline failed to develop and instead only a single fused telencephalic vesicle resulted. Sonic hedgehog still plays a role in differentiation, proliferation, and maintenance of adult tissues. Abnormal activation of SHH signaling in adult tissues has been implicated in various types of cancers including breast, skin, brain, liver, gallbladder and many more.

Background References

1. Zhang X et al. Expression of SOX9 and CDX2 in nongoblet columnar-lined esophagus predicts the detection of Barrett\'s esophagus during follow-up. Mod Pathol 28:654-61 (2015).

2. Li H et al. Olfactomedin 4 deficiency promotes prostate neoplastic progression and is associated with upregulation of the hedgehog-signaling pathway. Sci Rep 5:16974 (2015).

Sequence Similarity

Belongs to the hedgehog family.

Post-translational Modification

[Sonic hedgehog protein]: The C-terminal domain displays an autoproteolysis activity and a cholesterol transferase activity (By similarity). Both activities result in the cleavage of the full-length protein and covalent attachment of a cholesterol moiety to the C-terminal of the newly generated N-terminal fragment (ShhN) (By similarity). Cholesterylation is required for the sonic hedgehog protein N-product targeting to lipid rafts and multimerization. ShhN is the active species in both local and long-range signaling, whereas the C-product (ShhC) is degraded in the reticulum endoplasmic (By similarity).; [Sonic hedgehog protein N-product]: N-palmitoylation by HHAT of ShhN is required for sonic hedgehog protein N-product multimerization and full activity (By similarity). It is a prerequisite for the membrane-proximal positioning and the subsequent shedding of this N-terminal peptide.; [Sonic hedgehog protein N-product]: The lipidated N- and C-terminal peptides of ShhNp can be cleaved (shedding)(PubMed:24522195). The N-terminal palmitoylated peptide is cleaved at the Cardin-Weintraub (CW) motif site. The cleavage reduced the interactions with heparan sulfate. The cleavage is enhanced by SCUBE2.

Subcellular Location

Secreted, Cell membrane.

UNIPROT

SwissProt: Q15465 Human

SwissProt: Q62226 Mouse

SwissProt: Q63673 Rat

Synonyms

HHG 1 antibody

HHG-1 antibody

HHG1 antibody

HLP 3 antibody

HLP3 antibody

Holoprosencephaly 3 antibody

HPE 3 antibody

HPE3 antibody

MCOPCB5 antibody

shh antibody

Expand

Images

  • Western blot analysis of Sonic Hedgehog Protein/SHH(C-Product) on different lysates with Rabbit anti-Sonic Hedgehog Protein/SHH(C-Product) antibody (<a href="/products/ET1610-6" style="font-weight: bold;text-decoration: underline;">ET1610-6</a>) at 1/1,000 dilution.<br /><br />Lane 1: Mouse kidney tissue lysate (30 µg/Lane)<br />Lane 2: Mouse brain tissue lysate (30 µg/Lane)<br />Lane 3: Mouse liver tissue lysate (30 µg/Lane)<br />Lane 4: H22 cell lysate (15 µg/Lane)<br />Lane 5: Neuro-2a cell lysate (15 µg/Lane)<br />Lane 6: NIH/3T3 cell lysate (15 µg/Lane)<br /><br />Predicted band size: 50 kDa<br />Observed band size: 28/55 kDa<br /><br />Exposure time: 30 seconds;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1610-6" style="font-weight: bold;text-decoration: underline;">ET1610-6</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/100,000 dilution was used for 1 hour at room temperature.

    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: 28/55 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.

  • Western blot analysis of Sonic Hedgehog Protein/SHH(C-Product) on different lysates with Rabbit anti-Sonic Hedgehog Protein/SHH(C-Product) antibody (<a href="/products/ET1610-6" style="font-weight: bold;text-decoration: underline;">ET1610-6</a>) at 1/1,000 dilution.<br /><br />Lane 1: Rat heart tissue lysate<br />Lane 2: Rat liver tissue lysate<br />Lane 3: Rat brain tissue lysate<br />Lane 4: Rat lung tissue lysate<br />Lane 5: Mouse liver tissue lysate<br /><br />Lysates/proteins at 30 µg/Lane.<br /><br />Predicted band size: 50 kDa<br />Observed band size: 28/55 kDa<br /><br />Exposure time: 59 seconds; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1610-6" style="font-weight: bold;text-decoration: underline;">ET1610-6</a>) at 1/1,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    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: Rat heart tissue lysate
    Lane 2: Rat liver tissue lysate
    Lane 3: Rat brain tissue lysate
    Lane 4: Rat lung tissue lysate
    Lane 5: Mouse liver tissue lysate

    Lysates/proteins at 30 µg/Lane.

    Predicted band size: 50 kDa
    Observed band size: 28/55 kDa

    Exposure time: 59 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 (ET1610-6) at 1/1,000 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.

  • Western blot analysis of Sonic Hedgehog Protein on different lysates using anti-Sonic Hedgehog Protein antibody at 1/1,000 dilution.<br /><span style="font-weight: bold;">Positive control:</span> <br />Lane 1: Hela <br />Lane 2: HepG2

    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

  • Immunocytochemistry analysis of Neuro-2a cells labeling Sonic Hedgehog Protein/SHH(C-Product) with Rabbit anti-Sonic Hedgehog Protein/SHH(C-Product) antibody (<a href="/products/ET1610-6" style="font-weight: bold;text-decoration: underline;">ET1610-6</a>) at 1/100 dilution.<br /><br />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 (<a href="/products/ET1610-6" style="font-weight: bold;text-decoration: underline;">ET1610-6</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    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.

  • 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.

    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.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-Sonic Hedgehog Protein/SHH(C-Product) antibody (<a href="/products/ET1610-6" style="font-weight: bold;text-decoration: underline;">ET1610-6</a>) at 1/1,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1610-6" style="font-weight: bold;text-decoration: underline;">ET1610-6</a>) 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.

    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.

  • Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-Sonic Hedgehog Protein/SHH(C-Product) antibody (<a href="/products/ET1610-6" style="font-weight: bold;text-decoration: underline;">ET1610-6</a>) at 1/1,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1610-6" style="font-weight: bold;text-decoration: underline;">ET1610-6</a>) 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.

    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.

  • Immunohistochemical analysis of paraffin-embedded rat kidney tissue using anti-Sonic Hedgehog Protein antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded rat kidney tissue using anti-Sonic Hedgehog Protein antibody. Counter stained with hematoxylin.

  • Immunohistochemical analysis of paraffin-embedded rat liver tissue using anti-Sonic Hedgehog Protein antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded rat liver tissue using anti-Sonic Hedgehog Protein antibody. Counter stained with hematoxylin.

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