FZD7 antibody (AA 501-574)

Catalog No. ABIN710051

Product Details anti-FZD7 Antibody

Target: FZD7 (Frizzled Family Receptor 7 (FZD7))

Binding Specificity: AA 501-574

Reactivity: Human, Mouse, Rat

Host: Rabbit

Clonality: Polyclonal

Conjugate: This FZD7 antibody is un-conjugated

Application: Western Blotting (WB), ELISA, Flow Cytometry (FACS), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunofluorescence (Cultured Cells) (IF (cc)), Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunohistochemistry (Frozen Sections) (IHC (fro))

Cross-Reactivity: Human, Mouse, Rat

Predicted Reactivity: Dog,Cow,Pig,Chicken,Guinea Pig

Purification: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human Frizzled 7

Isotype: IgG

Application Details

Application Notes: WB 1:300-5000
ELISA 1:500-1000
FCM 1:20-100
IHC-P 1:200-400
IHC-F 1:100-500
IF(IHC-P) 1:50-200
IF(IHC-F) 1:50-200
IF(ICC) 1:50-200

Restrictions: For Research Use only

Independent Validation (WB)

Validation #029805 (Western Blotting)

by: Alamo Laboratories Inc

No.: #029805

Date: 08/26/2014

Lot Number: 120426

Method validated: Western Blotting

Positive Control: MDA-MB-231 cells

Negative Control: C6/36 cells (non-reactive species)

Notes: A strong band was observed in the positive control sample at the expected molecular weight of ~64 kDa. The band is also faintly observed in the negative control sample, which may indicate species cross-reactivity.

Validation Images

Figure 1: Western Blot for FZD7. Arrowhead indicates the expected molecular weight of ~64 kDa.

Full Methods

Primary Antibody:

• Antigen: Frizzled Family Receptor 7 (FZD7)
• Catalog number: ABIN710051
• Lot number: 120426
• Antibody Dilution: 1:100

Secondary Antibody:

• Antigen: Goat Anti-Rabbit IgG (H + L)-HRP Conjugate
• Lot number: L170-6515
• Antibody Dilution: 1:10,000

Full Protocol:

• 1. The cell extracts were heated at 95°C for 5 minutes in 1X SDS Sample Buffer containing 1% SDS and 1.25% β-mercaptoethanol.
• 2. 15 μl of heated culture-media were loaded and resolved on 8-16% SDS-polyacrylamide gel.
• 3. The Thermo Scientific - Spectra Multicolor Broad Range (Cat # 26634) were used as molecular mass markers.
• 4. Proteins were then transferred onto PVDF membrane by wet transfer and protein transfer was confirmed with Ponceau-S staining.
• 5. The PVDF membrane was incubated with 25 ml of blocking buffer [Tris Buffered Saline, pH 7.4 plus 0.1% TW20 (TBST)] containing 5% (W/V) BSA at room temperature for 1 hour.
• 6. The membrane was rinsed with TBST once.
• 7. The membrane was immersed with the protein side up in the primary antibody solution in TBST containing 5% (W/V) BSA and incubated for 24 hours at 4°C.
• 8. The membrane was rinsed in TBST thrice for 5 minutes each.
• 9. The membrane was incubated in the HRP-conjugated secondary antibody solution in TBST containing 5% (W/V) BSA and incubated for 1 hour at room temperature (~26°C) with gentle agitation.
• 10. The membrane was rinsed thrice TBST thrice for 5 minutes each.
• 11. The membrane was rinsed in TBS twice for 30 seconds each.
• 12. Signals were detected with ECL-2 Substrate. The blot was scanned for 45 minutes.
• 13. The membrane was rinsed three times TBST.
• 14. Incubated in Acidic Glycine Stripping Buffer at room temperature with gentle agitation for 3 times, 10 minutes each.
• 15. The membrane was washed in TBST 2 times for 10 minutes each.
• 16. Repeated Steps 5-12 with the loading control antibody (for Anti-actin) and its matching secondary antibody.

Experimental Notes: - No experimental challenges noted.

Handling

Format: Liquid

Concentration: 1 μg/μL

Buffer: 0.01M TBS( pH 7.4) with 1 % BSA, 0.02 % Proclin300 and 50 % Glycerol.

Preservative: ProClin

Precaution of Use: This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.

Storage: 4 °C,-20 °C

Storage Comment: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Expiry Date: 12 months

References for anti-FZD7 antibody (ABIN710051)

Deng, Zhang, Zhang, Wen, Miao, Guo: "MicroRNA-142-3p inhibits cell proliferation and invasion of cervical cancer cells by targeting FZD7." in: Tumour biology, (2015) (PubMed).

Target Details for FZD7

Target: FZD7 (Frizzled Family Receptor 7 (FZD7))

Alternative Name: Frizzled 7 (FZD7 Products)

Synonyms: FzE3 antibody, Fz-7 antibody, Xfz7 antibody, frizzled-7 antibody, frizzled7 antibody, frz-7 antibody, frz7 antibody, fz7 antibody, fzd7-a antibody, fzd7-b antibody, fze3 antibody, Fz7 antibody, cFz-7 antibody, fb38g02 antibody, fc44b09 antibody, fz13 antibody, fz7a antibody, fz7b antibody, wu:fb38g02 antibody, wu:fc44b09 antibody, zg13 antibody, frizzled class receptor 7 antibody, frizzled class receptor 7 L homeolog antibody, frizzled class receptor 7b antibody, FZD7 antibody, fzd7.L antibody, fzd7 antibody, Fzd7 antibody, fzd7b antibody

Background:

Synonyms: FzE3, Frizzled-7, Fz-7, hFz7, FZD7

Background: Receptor for Wnt proteins. Most of frizzled receptors are coupled to the beta-catenin canonical signaling pathway, which leads to the activation of disheveled proteins, inhibition of GSK-3 kinase, nuclear accumulation of beta-catenin and activation of Wnt target genes. A second signaling pathway involving PKC and calcium fluxes has been seen for some family members, but it is not yet clear if it represents a distinct pathway or if it can be integrated in the canonical pathway, as PKC seems to be required for Wnt-mediated inactivation of GSK-3 kinase. Both pathways seem to involve interactions with G-proteins. May be involved in transduction and intercellular transmission of polarity information during tissue morphogenesis and/or in differentiated tissues.

Gene ID: 8324

UniProt: O75084

Pathways: WNT Signaling, Stem Cell Maintenance