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Smoothened Homolog (Drosophila) (SMO) (Extracellular Domain), (N-Term) antibody

Details for Product No. ABIN1049353, Supplier: Login to see
Antigen
  • XSmo
  • smoothened
Epitope
N-Term, Extracellular Domain
8
5
2
2
1
1
1
1
1
1
1
Reactivity
Human
31
6
2
1
Host
Rabbit
24
6
2
Clonality
Polyclonal
Conjugate
Un-conjugated
1
1
1
1
1
1
Application
Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))
26
22
14
8
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Immunogen Synthetic 15 amino acid peptide from N-terminal extracellular domain of human SMO. Percent identity with other species by BLAST analysis: Human, Gorilla, Gibbon, Monkey, Marmoset, Mouse, Rat, Bovine, Dog, Bat, Hamster, Elephant, Panda, Horse, Rabbit, Pig, Platypus (100%), Opossum (93%), Turkey, Chicken, Xenopus (87%), Zebrafish, Seq squirt (80%).
ImmunogenType: Synthetic peptide
Specificity Human SMO. BLAST analysis of the peptide immunogen showed no homology with other human proteins.
Cross-Reactivity Bat, Cow (Bovine), Dog (Canine), Hamster, Horse (Equine), Monkey, Mouse (Murine), Pig (Porcine), Rabbit, Rat (Rattus)
Purification Immunoaffinity Chromatography
Alternative Name SMO / Smoothened (SMO Antibody Abstract)
Background Standard Gene Symbol: SMO, Gene Family: GPCR, Gene Subfamily: Frizzled, Synonyms: SMO, Frizzled family member 11, FZD11, Gx, Protein Gx, SMOH, Smoothened, Smoothened homolog
Gene ID 6608
Pathways
Application Notes IHC-P (10 μg/mL),
Immunohistochemistry: LS-A2666 was validated for use in immunohistochemistry on a panel of 21 formalin-fixed, paraffin-embedded (FFPE) human tissues after heat induced antigen retrieval in pH 6.0 citrate buffer. After incubation with the primary antibody, slides were incubated with biotinylated secondary antibody, followed by alkaline phosphatase-streptavidin and chromogen. The stained slides were evaluated by a pathologist to confirm staining specificity. The optimal working concentration for LS-A2666 was determined to be 10 μg/mL.
Comment

This antibody has been further validated specifically for use under standardized IHC conditions against formalin-fixed paraffin-embedded human tissues.

Assay Procedure

The IHC-pro Immunohistochemistry Protocol

Tissue Preparation
Formalin fixation and embedding in paraffin wax

Tissue Sectioning
Make 4-µm sections and place on pre-cleaned and charged microscope slides.
Heat in a tissue-drying oven for 45 minutes at 60°C
Deparaffinization
Wash slides in 3 changes of xylene – 5 minutes each at room temperature.

Rehydration
Wash slides in 3 changes of 100% alcohol – 3 minutes each at room temperature.
Wash slides in 2 changes of 95% alcohol – 3 minutes each at room temperature.
Wash slides in 1 change of 80% alcohol – 3 minutes at room temperature.
Rinse slides in gentle running distilled water – 5 minutes at room temperature.

Antigen retrieval
Steam slides in 0.01 M sodium citrate buffer, pH 6.0 at 99-100°C - 20 minutes
Remove from heat and let stand at room temperature in buffer - 20 minutes
Rinse in 1X TBS with Tween (TBST) – 1 minute at room temperature.

Immunostaining
Do not allow tissues to dry at any time during the staining procedure.
Apply a universal protein block – 20 minutes at room temperature.
Drain protein block from slides, apply diluted primary antibody – 45 minutes at room temperature.
Rinse slides in 1X TBST - 1 minute at room temperature.
Apply a biotinylated secondary antibody (specific to the host of the primary antibody) - 30 minutes at room temperature.
Rinse slides 1X TBST – 1 minute at room temperature.
Apply alkaline phosphatase streptavidin – 30 minutes at room temperature.
Rinse slides in 1X TBST - 1 minute at room temperature.
Apply alkaline phosphatase chromogen substrate – 30 minutes at room temperature.
Wash slides in distilled water – 1 minute at room temperature.

Dehydrate
This method should only be used if the chromogen substrate is alcohol insoluble.
Wash slides in 2 changes of 80% alcohol – 1 minute each at room temperature.
Wash slides in 2 changes of 95% alcohol – 1 minute each at room temperature.
Wash slides in 3 changes of 100% alcohol – 1 minute each at room temperature.
Wash slides in 3 changes of xylene – 1 minute each at room temperature.
Apply coverslip

Restrictions For Research Use only
Format Liquid
Concentration 1 mg/mL
Buffer PBS, less than 0.1 % sodium azide.
Preservative Sodium azide
Precaution of Use This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage 4 °C/-80 °C
Storage Comment Long term: -70°C, Short term: +4°C
Supplier Images
Immunohistochemistry (IHC) image for anti-Smoothened Homolog (Drosophila) (SMO) (Extracellular Domain), (N-Term) antibody (ABIN1049353) Human Kidney: Formalin-Fixed Paraffin-Embedded (FFPE)
Background publications Corbit, Aanstad, Singla et al.: "Vertebrate Smoothened functions at the primary cilium." in: Nature, Vol. 437, Issue 7061, pp. 1018-21, 2005 (PubMed).