Fianostics FluoBolt™ Wnt3A protein immunoassay is a 96-well format, complete kit with a standard and control samples in a pooled serum matrix.    This assay is similar to a standard ELISA in the fact that this assay is a sandwich immunoassay comprised of a monoclonal capture antibody and a polyclonal detection antibody, but different from a standard ELISA in the fact that it utilizes Metal Enhanced Fluorescence (MEF) to dramatically increase the sensitivity with the same antibodies.  MEF technology uses fluorescence detection that can enhance the signal more than 100 times over chromogenic or chemiluminescent signals.

Although there are assays for measuring WNT3A transcript levels available in the market, detection methods for protein levels have been limited and thus hamper Wnt3A research progress.  Existing options for measuring Wnt3A protein either require expensive closed-systems instruments, have inconsistent quality records, or are only semi-quantitative in nature (eg. Western blotting).  FluoBolt™-Technology to provides a high sensitivity Wnt3A protein assay for basic and clinical research, which increases detection and may improve data consistency better suited for research needs.

Read more about the MEF technology here.

The FluoBolt™ Wnt3A assay detects human Wnt3A protein. Human WNT3A shares between 100-95% aa sequence identity with primates, mice, rats, hamster, and pig. Cross-reactivity of this assay with other species than human has not been tested with FluoBolt™ Wnt3A.

 

Amino Acid Sequence Identity with Human
Source uniprot.org
Mouse Rat NHP Pig Canine Zebra Fish Hamster
Periostin 90% 87% 92-93% 95% 96% 54% 89%
Asporin 89% 89% 96-99% 89% 86% 54% 89%
Noggin 99% 99% 99-100% 99% 96% 55% 99%
Wnt3a 96% 89% 99-100% 96% 71% 82% 96%
Klotho 86% 84% 93-99% 88% n.a. 50% 85%

 

  • 1 x 96 well- Anti-human Wnt3A monoclonal antibody, pre-coated MEF-microtiter plate
  • 1 x 25 ml- Wash buffer concentrate 20x, natural cap
  • 1 x 2.5 ml- Anti-human Wnt3A polyclonal antibody, labeled with either: FITC, Cy3, Cy5 or AlexaFluor680
  • 6 vials, 0.25 ml- Standards 1-6, (2800, 1400, 700, 350, 175, 0 pmol/l),white caps, lyophilized

Standards are highly purified recombinant protein in human pooled-serum matrix.

  • 2 vials, 0.25 ml- Control A and B, yellow cap, lyophilized (for concentrations see label).
  • 1 x 10 ml- Sample diluent, natural cap, ready to use

 

ADDITIONAL MATERIAL SUPPLIED WITH THE KIT

  • 2 self-adhesive plastic films
  • QC data sheet
  • Protocol sheet
  • Instruction manual for use
  • 2 desiccant bags for plate storage

 

MATERIAL AND EQUIPMENT REQUIRED BUT NOT SUPPLIED

  • Precision pipettes calibrated to deliver 10 μl, 20 μl, 50 μl, 200 μl, 500 μl, and disposable tips
  • Distilled or deionized water
  • Plate washer, multichannel pipette, or manifold dispenser for washing
  • Refrigerator with 4°C (2-8°C)
  • Fluorescence microplate reader
Method Metal Enhanced Direct Sandwich Fluorescence Immunoassay in 96-well format
Sample Type Serum and Plasma
Standard Range 0 to 2,800 pmol/L (0-112 ng/mL)

6 standards and 2 controls in a serum based matrix

Conversion Factor 1 ng/mL = 25 pmol/L
Sample Volume 20 uL (undiluted sample)/well
Incubation/time/temperature Single-step assay / Overnight / Room Temperature
Sensitivity LOD (0 pmol/L+3 SD); 50 pmol/L (2 ng/mL)

LLOQ 175 pmol/L (7 ng/mL)

Specificity Tested for human WNT3A, but human shares 100-96% aa sequence homology with primates and mice.

Marketing Literature:  FluoBolt WNT3A Flyer (PDF)

Publications about Wnt3a:

  • Wnt3a: Functions and Implications in Cancer. Sha H et al., Chin J Cancer. 2015;34(12):554-62.
  • Wnt3a signaling within bone inhibits multiple myeloma bone disease and tumor growth. Qiang YW et al., Blood. 2008;112(2):374-382.
  • Therapeutic Potential of Wnt-3a in Neurological Recovery after Spinal Cord Injury.  Gao K, Zhang T, Wang F, Lv C: Eur Neurol. 2019;81:197-204.