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Sonic Hedgehog (Shh) Rabbit pAb (A7726)

Tested applications:WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP InhibitionReactivity:Human, Mouse, Rat

Western blot analysis of lysates from wild type (WT) and Sonic Hedgehog (Sonic Hedgehog (Shh)) knockout (KO) HeLa cells, using [KO Validated] Sonic Hedgehog (Shh) Rabbit pAb (A7726) at 1:1000 dilution.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 10s.

Western blot analysis of various lysates using Sonic Hedgehog (Shh) Rabbit pAb (A7726) at 1:1000 dilution.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 180s.

Western blot analysis of lysates from Rat lung, using Sonic Hedgehog (Shh) Rabbit pAb (A7726) at 1:1000 dilution.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Enhanced Kit (RM00021).
Exposure time: 180s.

ABclonal:Immunohistochemistry - Sonic Hedgehog (Shh) Rabbit pAb (A7726)

Immunohistochemistry analysis of paraffin-embedded Rat kidney using Sonic Hedgehog (Shh) Rabbit pAb (A7726) at dilution of 1:100 (40x lens). Microwave antigen retrieval performed with 0.01M PBS Buffer (pH 7.2) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Rat lung using Sonic Hedgehog (Shh) Rabbit pAb (A7726) at dilution of 1:100 (40x lens). Microwave antigen retrieval performed with 0.01M PBS Buffer (pH 7.2) prior to IHC staining.

ABclonal:Immunofluorescence - Sonic Hedgehog (Shh) Rabbit pAb (A7726)

Immunofluorescence analysis of HeLa cells using Sonic Hedgehog (Shh) Rabbit pAb (A7726) at dilution of 1:50 (40x lens). Secondary antibody: Cy3-conjugated Goat anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.

Immunofluorescence analysis of paraffin-embedded rat liver using Sonic Hedgehog (Shh) Rabbit pAb (A7726) at dilution of 1:50 (40x lens). Secondary antibody: Cy3-conjugated Goat anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.

Overview

Product name	Sonic Hedgehog (Shh) Rabbit pAb
Catalog No.	A7726
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG

Background

This gene encodes a protein that is instrumental in patterning the early embryo. It has been implicated as the key inductive signal in patterning of the ventral neural tube, the anterior-posterior limb axis, and the ventral somites. Of three human proteins showing sequence and functional similarity to the sonic hedgehog protein of Drosophila, this protein is the most similar. The protein is made as a precursor that is autocatalytically cleaved; the N-terminal portion is soluble and contains the signalling activity while the C-terminal portion is involved in precursor processing. More importantly, the C-terminal product covalently attaches a cholesterol moiety to the N-terminal product, restricting the N-terminal product to the cell surface and preventing it from freely diffusing throughout the developing embryo. Defects in this protein or in its signalling pathway are a cause of holoprosencephaly (HPE), a disorder in which the developing forebrain fails to correctly separate into right and left hemispheres. HPE is manifested by facial deformities. It is also thought that mutations in this gene or in its signalling pathway may be responsible for VACTERL syndrome, which is characterized by vertebral defects, anal atresia, tracheoesophageal fistula with esophageal atresia, radial and renal dysplasia, cardiac anomalies, and limb abnormalities. Additionally, mutations in a long range enhancer located approximately 1 megabase upstream of this gene disrupt limb patterning and can result in preaxial polydactyly.

Immunogen information

Immunogen	A synthetic peptide corresponding to a sequence within amino acids 200-300 of human Sonic Hedgehog (Shh) (NP_000184.1).
Sequence	PGSATVHLEQGGTKLVKDLSPGDRVLAADDQGRLLYSDFLTFLDRDDGAKKVFYVIETREPRERLLLTAAHLLFVAPHNDSATGEPEASSGSGPPSGGALGPRALFASRVRPGQRVYVVAERDGDRRLLPAAVHSVTLSEEAAGAYAPLTAQGTILINRVLASCYAVIEEHSWAHRAFAPFRLAHALLAALAPARTDRGGDSGGGDRGGGGGRVALTAPGAADAPGAGATAGIHWYSQLLYQIGTWLLDSEALHPLGMAVKSS
Gene ID
Swiss Prot
Synonyms	TPT; HHG1; HLP3; HPE3; SMMCI; ShhNC; TPTPS; MCOPCB5; Sonic Hedgehog (Shh)
Calculated MW	50kDa
Observed MW	19kDa

Applications

Reactivity	Human, Mouse, Rat
Tested applications	WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP Inhibition
Recommended dilution	WB 1:500 - 1:1000 IHC-P 1:50 - 1:100 IF/ICC 1:50 - 1:200 ELISA Recommended starting concentration is 1 μg/mL. Please optimize the concentration based on your specific assay requirements.
Storage buffer	Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.05% proclin300, 50% glycerol, pH7.3.
Key application	Western blotting Immunohistochemistry Immunofluorescence
Positive samples	HeLa, HepG2, Rat lung
Cellular location	Cell membrane, Lipid-anchor, Secreted, extracellular space.
Customer validation	WB(Rattus norvegicus, Homo sapiens, Mus musculus) IHC(Rattus norvegicus)