This Mouse Resistin ELISA kit is used for the non-radioactive quantification of Mouse Resistin in mouse serum, plasma, adipocyte extracts or cell culture media samples. This kit has 100% cross reactivity to Mouse Resistin. One kit is sufficient to measure 38 unknown samples in duplicate. This kit is for research purpose only.



This assay is a Sandwich ELISA based, sequentially, on: 1) capture of Mouse Resistin molecules from samples to the wells of a microtiter plate coated by a pre-titered amount of anti-mouse resistin monoclonal antibodies, 2) wash away of unbound materials from samples, 3) binding of a second biotinylated anti-mouse polyclonal antibody to the captured molecules, 4) wash away of unbound materials from samples, 5) conjugation of horseradish peroxidase to the immobilized biotinylated antibodies, 6) wash away of free enzyme conjugates, and 7) quantification of immobilized antibody-enzyme conjugates by monitoring horseradish peroxidase activities in the presence of the substrate 3,3’,5,5’-tetramethylbenzidine. The enzyme activity is measured spectrophotometrically by the increased absorbency at 450 nm, corrected from the absorbency at 590nm, after acidification of formed products. Since the increase in absorbency is directly proportional to the amount of captured Mouse Resistin in the unknown sample, the latter can be derived by interpolation from a reference curve generated in the same assay with reference standards of known concentrations of Mouse Resistin.



Each kit is sufficient to run one 96-well plate and contains the following reagents:


A. Mouse Resistin ELISA Plate

Coated with Monoclonal anti-Mouse Resistin Antibodies

Quantity: 1 plate

Preparation: Ready to Use

B. Adhesive Plate Sealer

Quantity: 2 sheet

Preparation: Ready to Use

C. 10X HRP Wash Buffer Concentrate

10X concentrate of 50mM Tris Buffered Saline containing Tween-20.

Quantity: 2 bottles containing 50 ml each

Preparation: Dilute 1:10 with distilled or deionized water.

D. Mouse Resistin Standard

Purified Recombinant Mouse Resistin, 50 ng lyophilized.

Quantity: 1ml upon hydration

Preparation: Contents Lyophilized. Reconstitute with 1 ml distilled or deionized water.


E. Mouse Resistin Quality Controls 1 and 2

Purified Recombinant Mouse Resistin, lyophilized.

Quantity: 1ml/vial upon hydration

Preparation: Contents Lyophilized. Reconstitute with 1 ml distilled or deionized water.

F. Assay Buffer

Buffer containing BSA and 0.08% Sodium Azide

Quantity: 20 ml

Preparation: Ready to Use

G. Mouse Resistin Detection Antibody

Pre-titered Biotinylated Goat anti-Mouse Resistin Polyclonal Antibody

Quantity: 6 ml

Preparation: Ready to Use

H. Enzyme Solution

Pre-titered Streptavidin-Horseradish Peroxidase Conjugate in Buffer

Quantity: 12 ml

Preparation: Ready to Use

I. Substrate (Light sensitive, avoid unnecessary exposure to light)

3, 3’, 5, 5’-tetramethylbenzidine in buffer

Quantity: 12 ml

Preparation: Ready to Use.

J. Stop Solution (Caution: Corrosive Solution)

0.3 M HCl

Quantity: 12 ml

Preparation: Ready to Use

K. Matrix Solution

Quantity: 1 ml/vial

Preparation: Ready to Use

Prior to use, all components in the kit can be stored up to 2 weeks at 2-8oC. For longer storage (> 2 weeks), freeze Wash Buffer, Assay Buffer, Serum Matrix, Resistin Standards, Quality Controls and reconstituted Standards and Controls at £ –20oC. Minimize repeated freeze and thaw of the Resistin Standards and Quality Controls. Refer to expiration dates on all reagents prior to use. Do not mix reagents from different kits unless they have the same lot numbers.




A. Sodium Azide

Sodium azide has been added to certain reagents as a preservative. Although the concentrations are low, sodium azide may react with lead and copper plumbing to form highly explosive metal azides. On disposal, flush with a large volume of water to prevent azide build up.


B. Hydrochloric Acid

Hydrochloric Acid is corrosive and can cause eye and skin burns. It is harmful if swallowed and can cause respiratory and digestive tract burns. Avoid contact with skin and eyes. Do not swallow or ingest.


  1. Pipettes and Pipette Tips: 10m l - 20 m l or 20m l - 100 m l
  2. Multi-Channel Pipettes and Pipette Tips: 5 ~ 50 µl and 50 ~ 300 m l
  3. Buffer and Reagent Reservoirs
  4. Vortex Mixer
  5. Deionized Water
  6. Microtiter Plate Reader capable of reading absorbency at 450 nm
  7. Orbital Microtiter Plate Shaker
  8. Absorbent Paper or Cloth



    1. To prepare serum samples, whole blood is directly drawn into a centrifuge tube that contains no anti-coagulant. Let blood clot at room temperature for 30 min.

Promptly centrifuge the clotted blood at 2,000 to 3,000 x g for 15 minutes at 4 ± 2oC.

Transfer and store serum samples in separate tubes. Date and identify each sample.

Use freshly prepared serum or aliquot and store samples at £ –20oC for later use. For long-term storage, keep at -70 oC. Avoid freeze/thaw cycles.


  1. To prepare plasma samples, whole blood should be collected into centrifuge tubes containing enough K3EDTA to achieve a final concentration of 1.735 mg/ml and centrifuged immediately after collection. Observe the same precautions in the preparation of serum samples.
  2. If heparin is to be used as an anticoagulant, the effect on the assay outcome at the dose of heparin used should be pre-determined.
  3. Avoid using samples with gross hemolysis or lipemia.



  1. No dilution or preparation is needed for normal serum or plasma samples. In the event that any sample is above 50 ng/ml range, dilutions should be performed using the Serum Matrix provided.
  2. Adipocyte extracts or cell culture media samples may require dilution. Dilutions should be performed using the assay buffer provided.



  • Mouse Resistin Standard Preparation


1. Use care in opening the lyophilized Standard vial. Using an Eppendorf pipette, reconstitute the Mouse Resistin Standard with 1 ml distilled or deionized water into the glass vial to give a 50 ng/mL concentration of Standard. Invert and mix gently, let sit for 5 minutes then mix well.

2. Label six tubes 25, 12.5, 6.25, 3.125, 1.56, and 0.78ng/ml. Add 0.5 ml Assay Buffer to each of the six tubes. Prepare serial dilutions by adding 0.5 ml of the 50 ng/ml reconstituted standard to the 25 ng/ml tube, mix well and transfer 0.5 ml of the 25 ng/ml standard to the 12.5 ng/ml tube, mix well and transfer 0.5 ml of the 12.5 ng/ml standard to the 6.25 ng/ml tube, mix well and transfer 0.5 ml of the 6.25 ng/ml standard to the 3.125 ng/ml tube, mix well and transfer 0.5 ml of the 3.125 ng/ml standard to the 1.56 tube, mix well and transfer 0.5 ml of the 1.56 ng/ml standard to the 0.78 ng/ml tube and mix well.


Note: Do not use a Repeater pipette. Change tip for every dilution. Wet tip with Standard before dispensing. Unused portions of standard should be stored at £ -20°C. Avoid multiple freeze/thaw cycles.