SMO antibody (N-Term)

Catalog No. ABIN1049353

Product Details anti-SMO Antibody

Target: SMO (Smoothened Homolog (Drosophila) (SMO))

Binding Specificity: N-Term

Reactivity: Human, Mouse, Rat, Dog, Cow, Hamster, Horse, Monkey, Pig, Rabbit, Bat

Host: Rabbit

Clonality: Polyclonal

Conjugate: This SMO antibody is un-conjugated

Application: Western Blotting (WB), Immunohistochemistry (IHC), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunofluorescence (IF), Immunocytochemistry (ICC)

Brand: IHC-plus™

Specificity: Human SMO. BLAST analysis of the peptide immunogen showed no homology with other human proteins.

Purification: Immunoaffinity purified

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%).

Type of Immunogen: Synthetic peptide

Application Details

Application Notes: Approved: ICC, IF, IHC, IHC-P (10 μg/mL), WB

Usage: Immunohistochemistry: This antibody 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 this antibody was determined to be 10 μg/mL.

Comment:

Target Species of Antibody: Human

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

Handling

Format: Liquid

Concentration: Lot specific

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,-20 °C

Storage Comment: Aliquot and store undiluted at -20°C or below for up to 1 year. Can be stored undiluted at 4°C for up to 1 month. Avoid freeze-thaw cycles.

Expiry Date: 12 months

Target Details for SMO

Target: SMO (Smoothened Homolog (Drosophila) (SMO))

Alternative Name: SMO / Smoothened (SMO Products)

Synonyms: FZD11 antibody, Gx antibody, SMOH antibody, Smoh antibody, E130215L21Rik antibody, bnb antibody, smoothened antibody, XSmo antibody, smoothened, frizzled class receptor antibody, smoothened, frizzled class receptor L homeolog antibody, SMO antibody, Smo antibody, smo.L antibody, smo antibody

Background: Name/Gene ID: SMO
Subfamily: Frizzled
Family: GPCR

Synonyms: SMO, Frizzled family member 11, FZD11, Gx, Protein Gx, SMOH, Smoothened, Smoothened homolog

Gene ID: 6608

Pathways: Hedgehog Signaling