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SHH antibody - N-terminal region (ARP44235_P050)

100 ul

Regular Price: $289.00

Special Price: $215.00

In Stock

Conjugation Options

ARP44235_P050-FITC Conjugated

ARP44235_P050-HRP Conjugated

ARP44235_P050-Biotin Conjugated

Gene Symbol:
SHH
NCBI Gene Id:
6469
Official Gene Full Name:
Sonic hedgehog
Protein Name:
Sonic hedgehog protein
Swissprot Id:
Q15465
Protein Accession #:
NP_000184
Nucleotide Accession #:
NM_000193
Alias Symbols:
HHG1, HLP3, HPE3, SMMCI, TPT, TPTPS, MCOPCB5
Replacement Item:
This antibody may replace item sc-1194 from Santa Cruz Biotechnology.
Description of Target:
SHH is 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. Defects in this protein or in its signalling pathway are a cause of holoprosencephaly (HPE). It is also thought that mutations in its 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.This gene, which is expressed only during embryogenesis, 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. In addition, it is 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.
Protein Size (# AA):
462
Molecular Weight:
28kDa
Host:
Rabbit
Clonality:
Polyclonal
Purification:
Affinity Purified
Application:
IHC, WB
Tissue Tool:
Find tissues and cell lines supported by DNA array analysis to express SHH.
RNA Seq:
Find tissues and cell lines supported by RNA-seq analysis to express SHH.
Immunogen:
The immunogen is a synthetic peptide directed towards the N terminal region of human SHH
Species Reactivity:
Cow, Dog, Goat, Guinea Pig, Horse, Human, Mouse, Rabbit, Rat, Zebrafish, Chicken
Predicted Homology Based on Immunogen Sequence:
Cow: 93%; Dog: 100%; Goat: 93%; Guinea Pig: 100%; Horse: 100%; Human: 100%; Mouse: 100%; Rabbit: 100%; Rat: 100%; Zebrafish: 100%
Complete computational species homology data:
Anti-SHH (ARP44235_P050)
Peptide Sequence:
Synthetic peptide located within the following region: RCLLLVLVSSLLVCSGLACGPGRGFGKRRHPKKLTPLAYKQFIPNVAEKT
Product Format:
Liquid. Purified antibody supplied in 1x PBS buffer with 0.09% (w/v) sodium azide and 2% sucrose.
Reconstitution and Storage:
For short term use, store at 2-8C up to 1 week. For long term storage, store at -20C in small aliquots to prevent freeze-thaw cycles.
Concentration:
Batch dependent within range: 100 ul at 0.5 - 1 mg/ml
Protein Interactions:
UBC; SEL1L; DERL2; DERL1; SYVN1; HHIP; PTCH2; PTCH1; SHH; ADCYAP1; GAS1;
Blocking Peptide:
For anti-SHH (ARP44235_P050) antibody is Catalog # AAP44235 (Previous Catalog # AAPP25615)
Datasheets/Manuals:
Printable datasheet for anti-SHH (ARP44235_P050) antibody
Target Reference:
Coon,D.R., (2006) Exp. Mol. Pathol. 80 (2), 119-123
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1 Item(s)

02/01/2017 16:23
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Product Review: SHH antibody - N-terminal region (ARP44235_P050) in Human glioma cells using IHC
Product page for SHH antibody - N-terminal region (ARP44235_P050)

Researcher: ELIAS EL HABR, INSERM
Application: IHC
Species + Tissue/Cell type: Human glioma cells
Primary antibody dilution: 1:200
Secondary antibody: Anti-rabbit-GFP
Secondary antibody dilution: 1:500


Questionnaire:
How would you rate this antibody on a scale from 1-5 (5=best) and why? 4. Good staining.
Would you use this antibody in future experiments? Yes
Have you used another antibody which has worked in your application? Yes
Do you believe the information about the reagent on Aviva's website is correct? Yes
If the antibody works, do you plan to use it in future experiments or to publish your data? Why or why not? Yes. Since SHH is implicated in my research projects.
How did you store the antibody after re-suspension? Aliquots of 10uL stored at -20 degree C.
Sample Description (please include species type and tissue/cell type): Human glioma cells.
Please explain fixation solution/method used (formalin, periodate-lysine-paraformaldehyde, acetone, etc.)? Pure methanol for 20min at -20C
How many different experimental trials were conducted using the antibody sample? At least 3
Primary antibody dilution, incubation time and temperature: 1:200 overnight at +4C
Secondary antibody used, dilution, incubation time and temperature: Anti-Rabbit IgG (ab96919, Abcam)/1:500/1 hour/ room temperature
From your IHC/ICC images, briefly explain the colors of each stain and counterstain: In green (GFP) SHH and in blue DAPI (DNA)/ + merged figure
Did you use an antigen retrieval method? If so, please explain? NO
What controls were used in your experiment? NONE
Please include your detailed tissue preparation and staining procedure/protocol here: 1. Spot of cells (10.000) are deposed on a super-frost slide
2. After the spot is dry, the slide is incubated in frozen pure methanol for 20min at -20C
3. Slides are washed 3X in PBS1X and kept at +4C untill used
4. Before ICC, cells are permeabilised with saponine buffer (0,1% of 5% saponine diluted in HBSS) for 30min
5. Non-specific sites are blocked using a solution of 2.5% SVF, 3% BSA in PBS1X
6. Without washing, remove the blocking solution and incubate the spot with primary antibody diluted (see above) in a 0,3% BSA PBS1X solution overnight at +4C
7. Wash cell spot 3 times using PBS1X for 5 min
8. Incubate using the appropriate secondary antibody for 1 hour at RT
9. Wash cell spot 3 times using PBS1X for 5 min
10. Stain DNA using DAPI or TOPRO as recomended
11. Wash cell spot 3 times using PBS1X for 5 min and 1 time using PB1X for 5 min
12. Use the appropriate coverslip
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