MYL3/CMLC1 Kit ELISA

N° du produit ABIN956159

Ce kit ELISA Colorimetric est conçu pour la mesure quantitative de Rat et Souris MYL3/CMLC1.

Aperçu rapide pour MYL3/CMLC1 Kit ELISA (ABIN956159)

Antigène: MYL3/CMLC1 (MYL3) (Myosin, Light Chain 3 (MYL3))

Reactivité: Rat, Souris

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Application: ELISA

Type d'échantillon: Plasma, Serum

Détail du produit MYL3/CMLC1 Kit ELISA

Analytical Method: Quantitative

Attributs du produit: The antibodies used in this ELISA kit are approximately ten times more sensitive toward cardiac MLC-1 (CMLC-1). The assay provides an excellent tool for assessment of cardiac injury when skeletal muscle injury can be excluded or controlled.The Rat & Mouse Cardiac Myosin Light Chain-1 ELISA uses two different CMLC-1 monoclonal antibodies. One is used for solid phase immobilization (on the microtiter wells). The second is conjugated to horse radish peroxidase (HRP) and is used for detection. The sample is diluted with diluent as necessary and 100 µL aliquots of samples and calibrators are incubated in the microtiter wells for one hour. The wells are then washed and anti-CMLC-1 HRP conjugate is added and incubated in the wells for one hour. CMLC-1 molecules are thereby sandwiched between the solid phase and HRP-conjugated antibodies. After washing to remove unbound HRP conjugate a solution of tetramethylbenzidine (TMB), an HRP substrate, is then added to the wells and incubated for 20 minutes, resulting in the development of a blue color. Color development is stopped by the addition of 1N HCl changing the color to yellow. The concentration of CMLC-1 is proportional to the absorbance at 450 nm and is derived from a calibration curve.

Ingrédients: Anti-CMLC-1-coated microtiter wells, 96 wells (12 x 8-well strips)
CMLC-1 Calibrator (lyophilized)
Diluent, 25 mL
CMLC-1 HRP Conjugate, 11 mL
Wash Solution (20X), 50 mL
TMB Reagent, 11 mL
Stop Solution, 11 mL.

Matériel non inclus: Pipettes: P-10, P-200 & P-1000 or equivalent
Disposable pipette tips
Distilled or de-ionized water
Vortex mixer
Absorbent paper
Graph paper or appropriate PC graphing software
Polypropylene microcentrifuge tubes (1.5 mL)
Microtiter plate reader capable of reading 0 to 4 OD at 450 nm.

Information d'application

Plaque: Pre-coated

Préparation de l'échantillon:

Plasma and serum should be prepared as quickly as possible after blood collection and stored at 4°C. All samples should be similarly processed (i.e., storage times and temperatures should be the same for all samples). If samples cannot be assayed within 4 hours of collection they should be frozen at -70°C and thawed only once prior to use. We recommend that samples be assayed in duplicate. Optimum sample dilution should be determined by the end user. Samples should only be diluted with the diluent supplied with the kit.

Procédure de l'essai:

1. Secure the desired number of coated wells in the holder.
   2. Dispense 100 µL of calibrators and diluted samples into the appropriate wells.
   3. Thoroughly mix and incubate on an orbital micro-plate shaker at 150 rpm at room temperature (18-25°C) for one hour.
   4. Remove the incubation mixture using either a plate washer or by flicking plate contents into an appropriate Bio-waste container.
   5. Wash and empty the microtiter wells 5 times with 1x wash solution preferably using a plate washer (400 µL/well). Alternatively, a squirt bottle may be used. The entire wash procedure should be performed as quickly as possible.
   6. Strike the wells sharply onto absorbent paper or paper towels to remove all residual droplets.
   7. Add 100 µL of enzyme conjugate reagent into each well.
   8. Incubate on an orbital micro-plate shaker at 150 rpm at room temperature (18-25°C) for one hour.
   9. Wash as detailed in 4 to 6 above.
   10. Dispense 100 µL of TMB Reagent into each well.
   11. Gently mix on an orbital micro-plate shaker at ~150 rpm at room temperature (18-25°C) for 20 minutes.
   12. Stop the reaction by adding 100 µL of Stop Solution to each well.
   13. Gently mix. It is important to make sure that all the blue color changes to yellow.
   14. Read the optical density at 450 nm with a microtiter plate reader within 5 minutes. Due to plate reader differences, the high calibrator absorbance values may occasionally be out of range. If this occurs, absorbance values may be determined at 405 nm instead.
   15. If absorbance values of samples exceed those of the highest calibrator, samples should be further diluted with diluent and re-tested. For practical purposes, samples with absorbance values below that of the lowest calibrator should be assigned a zero value.

Calcul des résultats:

1. Calculate the mean absorbance value (A450) for the calibrators and samples.
   2. Construct a calibration curve by plotting the A450 values obtained for each reference calibrator against its concentration in ng/mL on graph paper, with absorbance on the vertical (y) axis and concentration on the horizontal (x) axis.
   3. Using the A450 values for each sample, determine the corresponding concentration of CMLC-1 (ng/mL) from the calibration curve. If using graphing software, we suggest using a linear, point-to-point, or two site binding (hyperbola) fit of the data. The end user should choose the best data fit for the calibration curve.
   4. Multiply the derived CMLC-1 concentrations by the dilution factor to obtain the actual CMLC-1 concentration.

Restrictions: For Research Use only

Stockage

Stock: 4 °C

Stockage commentaire: Store the kit at 4°C. Keep the microtiter plate in a sealed bag with desiccant to minimize exposure to damp air. The expiration date of the kit is indicated on the box label.

Date de péremption: The expiry date is stated on the label.

Détail du antigène

Antigène: MYL3/CMLC1 (MYL3) (Myosin, Light Chain 3 (MYL3))

Autre désignation: Cardiac Myosin Light Chain-1

Sujet: Myosin light chains are released into the circulation following muscle injury and provide useful biomarkers of muscle damage. Myosin light chain-1 (MLC-1) is expressed as different but immunologically related isoforms in cardiac and skeletal muscle.