Product nameAnti-Ihh antibody
See all Ihh primary antibodies
DescriptionRabbit polyclonal to Ihh
Tested applicationsSuitable for: WB, ICC/IF, IHC-Pmore details
Species reactivityReacts with: Mouse, Human
Predicted to work with: Cow
- Brain (Mouse) Tissue Lysate Colon (Mouse) Tissue Lysate - normal tissue NIH 3T3 (Mouse embryonic fibroblast cell line) Whole cell lysate
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab39634 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 42 kDa (predicted molecular weight: 45 kDa).|
|ICC/IF||Use a concentration of 1 µg/ml.|
|IHC-P||Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
FunctionIntercellular signal essential for a variety of patterning events during development. Binds to the patched (PTC) receptor, which functions in association with smoothened (SMO), to activate the transcription of target genes. Implicated in endochondral ossification: may regulate the balance between growth and ossification of the developing bones. Induces the expression of parathyroid hormone-related protein (PTHRP).
Tissue specificityExpressed in embryonic lung, and in adult kidney and liver.
Involvement in diseaseDefects in IHH are the cause of brachydactyly type A1 (BDA1) [MIM:112500]. BDA1 is an autosomal dominant disorder characterized by middle phalanges of all the digits rudimentary or fused with the terminal phalanges. The proximal phalanges of the thumbs and big toes are short.
Defects in IHH are a cause of acrocapitofemoral dysplasia (ACFD) [MIM:607778]. ACFD is a disorder characterized by short stature of variable severity with postnatal onset. The most constant radiographic abnormalities are observed in the tubular bones of the hands and in the proximal part of the femur. Cone-shaped epiphyses or a similar epiphyseal configuration with premature epimetaphyseal fusion result in shortening of the skeletal components involved. Cone-shaped epiphyses were also present to a variable extent at the shoulders, knees, and ankles.
Sequence similaritiesBelongs to the hedgehog family.
modificationsThe C-terminal domain displays an autoproteolysis activity and a cholesterol transferase activity. 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 (N-product). The N-product is the active species in both local and long-range signaling, whereas the C-product has no signaling activity.
Cholesterylation is required for N-product targeting to lipid rafts and multimerization.
Palmitoylated. N-palmitoylation is required for N-product multimerization and full activity.
Cellular localizationSecreted > extracellular space. The C-terminal peptide diffuses from the cell and Cell membrane. The N-terminal peptide remains associated with the cell surface.
- Information by UniProt
- BDA1 antibody
- HHG-2 antibody
- HHG2 antibody
ab39634 (1/50) staining Ihh in paraffin-embbeded mouse pancreas tissue sections. Tissue underwent fixation in formaldehyde, heat-mediated antigen retrieval in citrate buffer pH 6.0 and blocking (5 minutes/peroxidase block and 10 minutes/protein block). For further experimental details please refer to abreview. Strongest staining observed in islet cells of the pancreas.
Anti-Ihh antibody (ab39634) at 1 µg/ml + Lung (Human) Whole Cell Lysate - fetal normal tissue (ab30282) at 10 µg
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 45 kDa
Observed band size: 45 kDa
Exposure time: 30 seconds
All lanes : Anti-Ihh antibody (ab39634) at 1 µg/ml
Lane 1 : Brain (Mouse) Tissue Lysate
Lane 2 :
Mouse colon tissue lysate - total protein (ab29544)
Lane 3 :
NIH 3T3 whole cell lysate (ab7179)
Lysates/proteins at 10 µg per lane.
All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 45 kDa
Observed band size: 42 kDa why is the actual band size different from the predicted?
The Ihh protein has a predicted molecular weight of 45 kDa. The first 27 amino acids of the Ihh sequence act as a signal sequence, and when cleaved the protein has an expected molecular weight of 42 kDa.
IHC image of Ihh staining in human normal kidney formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab39634, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
ICC/IF image of ab39634 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab39634, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) HepG2 cells at 5µg/ml.
This product has been referenced in:
- Zuo C et al. SHP2 regulates skeletal cell fate by modifying SOX9 expression and transcriptional activity. Bone Res 6:12 (2018). Read more (PubMed: 29644115) »
- Rodriguez AM & Downs KM Visceral endoderm and the primitive streak interact to build the fetal-placental interface of the mouse gastrula. Dev Biol N/A:N/A (2017). IHC ; Mouse . Read more (PubMed: 28882402) »