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 Easy-Titer Antibody Assay Kits 

Fast and specific determination of mouse, rabbit or human IgG or IgM concentration.

Thermo Scientific Easy-Titer Antibody Assay Kits include antibody-sensitized microspheres to measure the specific concentration of mouse, rabbit and human antibodies by an easy and rapid microagglutination technique using standard microplates and UV-Vis plate reader (spectrophotometer). Each kit is specific for a particular species and class of immunoglobulin and, unlike total protein assays, can specifically measure the concentration of target antibody in samples (e.g., serum, plasma, culture supernatant) that contain other proteins. The kits are extremely sensitive, requiring very small sample volumes. Antibody concentration is determined from the assay response (absorbance) by comparison to a standard curve prepared using dilutions of a known antibody sample (sold separately).

Easy-Titer Assay Kit results plotted from 340 nm or 405 nm absorbance measurements.

Generalized standard curve for Thermo Scientific Easy-Titer Antibody Assay Kits. The typical assay range is 8 to 500ng/mL.

Highlights:

  • Kits for five popular targets – choose a kit specific for a particular species and class of immunoglobulin; no cross-reaction with other species and classes of the target antibody
  • Antibody-based specificity – measure concentration of target antibody in a sample, not just total protein; no need to purify antibody to assess its concentration
  • Faster and easier than ELISA – three-component, homogenous assay; 10 minutes total incubation time
  • No special equipment needed – uses standard vortex mixer, pipetter, 96-well microplate, plate shaker and reader (measure absorbance at 340nm or 400nm)
  • Sensitive – assay range (standard curve) 8 to 500ng/mL; requires starting sample of at least 300ng/mL
  • Reproducible – coefficient of variation < 5%; error depends on dilution and pipetting technique
  • Antibody standards sold separately – see product list for suggested products; use any antibody standard with proper target identity and known concentration (greater than 10µg/mL)

How Easy-Titer Antibody Assays Work

Easy-Titer Assay Kits detect and measure specific target antibodies using agglutination of microspheres that are coated ("sensitized") with the specific anti-IgG or IgM polyclonal antibodies. In the appropriate aqueous buffer (supplied in kit), the monodispersed antibody-coated microspheres (> 1µm diameter) have highest absorptivity (l-max) to incident light having a wavelength (340nm) that is equal to approximately half their diameter. When sample is added, two or more microspheres bind to each antibody target via their coated specific polyclonal antibodies, and this agglutination into effectively larger apparent spheres results in proportional decrease in absorptivity (lower absorbance).

Easy-Titer Assay Kits use microagglutination to specifically detect target antibodies.

Microsphere agglutination assay. Thermo Scientific Easy-Titer Assay Kits use antibody-sensitized microspheres to specifically assay target antibody species based on microagglutination.

Typical microagglutination assays depend on a change in light-scattering and corresponding change in transmittance, to which absorbance is inversely related. Easy-Titer Assay Kits use a special dilution buffer whose refractive index eliminates the effect of light-scattering on the monodispersed microspheres for the measurement wavelength used. Because of this, the final 10- to 20-fold dilution of the sample for use in the assay must be done using the Dilution Buffer supplied in the kit.

Easy-Titer Human IgG-Gamma Assay Kit does not cross-react with Fab fragments

Figure (above). Human IgG-Gamma Kit (#23325) specifically detects whole IgG (blue) or Fc fragments (red). Fab fragments are not detected. Because IgG has a larger mass than the other fragments assayed, its line appears relatively shifted upward (or right) in this plot of mass-per-volume concentrations.

 

Figure (right). Human IgG Kit (#23310) detects human IgG and does not cross-react with human IgM.

Easy-Titer Human IgG Assay Kit does not cross-react with IgM
Figure (right). Human IgM Kit (#23315) detects human IgG and does not cross-react with human IgG. Easy-Titer Human IgM Assay Kit does not cross-react with IgG

Easy-Titer Antibody Assays are faster and easier than ELISA:

  1. Prepare standards (5 to 500ng/mL) by diluting purified antibody in Kit Dilution Buffer
  2. Prepare samples by diluting at least 20-fold in Dilution Buffer to within assay range (8 to 500ng/mL).
  3. Vortex vial of microsphere beads to create homogeneous suspension.
  4. Pipette 20µL of bead suspension and 20µL of each sample and standard into 96-well microplate wells.
  5. Incubate microplate for 5 minutes with vigorous mixing.
  6. Add 100µL of Kit Blocking Reagent.
  7. Incubate microplate for 5 minutes with vigorous mixing.
  8. Measure absorbance on standard plate reader (340nm or 405nm).
  9. Plot standard curve and interpolate samples to determine concentration.


Ordering Information
Bulk quantities: InquireBulk quantities: Inquire    Certificate of AnalysisCertificate of Analysis   Product InstructionsProduct Instructions   MSDSMSDS
Product # Description Pkg. Size Files Price
23300 Easy-Titer Mouse IgG Assay Kit
Sufficient For: 96 tests (9 standards plus 87 test samples or replicates).
Formulation: Specific for all subclasses of mouse IgG.
Kit Contents:
Goat Anti-Mouse IgG Sensitized Beads, 2mL
Easy-Titer Dilution Buffer, 30mL
Easy-Titer Blocking Buffer, 15mL
96-test kit Product Instructions MSDS for product # 23300 Easy-Titer Mouse IgG Assay Kit $280.00

31202 Mouse IgG, Whole Molecule 5 mg MSDS for product # 31202 Mouse IgG, Whole Molecule $144.00

23305 Easy-Titer Rabbit IgG Assay Kit
Sufficient For: 96 tests (9 standards plus 87 test samples or replicates).
Formulation: Specific for all subclasses of rabbit IgG.
Kit Contents:
Goat Anti-Rabbit IgG Sensitized Beads, 2mL
Easy-Titer Dilution Buffer, 30mL
Easy-Titer Blocking Buffer, 15mL
96-test kit Product Instructions MSDS for product # 23305 Easy-Titer Rabbit IgG Assay Kit $278.00

31235 Rabbit IgG, Whole Molecule 10 mg $90.00

23310 Easy-Titer Human IgG (H+L) Assay Kit
Sufficient For: 96 tests (9 standards plus 87 test samples or replicates).
Formulation: Specific for all subclasses, fragments and chains of human IgG.
Kit Contents:
Goat Anti-Human IgG (H+L) Sensitized Beads, 2mL
Easy-Titer Dilution Buffer, 30mL
Easy-Titer Blocking Buffer, 15mL
96-test kit Product Instructions MSDS for product # 23310 Easy-Titer Human IgG (H+L) Assay Kit $303.00

31154 Human IgG, Whole Molecule
10 mg $130.00

23325 Easy-Titer Human IgG (gamma chain) Assay Kit
Sufficient For: 96 tests (9 standards plus 87 test samples or replicates).
Formulation: Specific for the Fc fragment of all subclasses of human IgG.
Kit Contents:
Goat Anti-Human IgG (gamma chain) Sensitized Beads, 2mL
Easy-Titer Dilution Buffer, 30mL
Easy-Titer Blocking Buffer, 15mL
96-test kit Product Instructions MSDS for product # 23325 Easy-Titer Human IgG (gamma chain) Assay Kit $315.00

23315 Easy-Titer Human IgM Assay Kit
Sufficient For: 96 tests (9 standards plus 87 test samples or replicates).
Formulation: Specific for all human IgM.
Kit Contents:
Goat Anti-Human IgM Sensitized Polystryrene Beads, 2mL
Easy-Titer Dilution Buffer, 30mL
Easy-Titer Blocking Buffer, 15mL
96-test kit Product Instructions MSDS for product # 23315 Easy-Titer Human IgM Assay Kit $313.00

31146 Human IgM (myeloma), whole molecule 2 mg MSDS for product # 31146 Human IgM (myeloma), whole molecule $143.00

15041 96-Well Plates — Corner Notch 100 plates/pkg $312.00


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