Salbutamol Elisa Kit-Food safety

Salbutamol Elisa Kit-Food Safety
 
INTENTED USE
Salbutamol ELISA Kit is a direct enzyme-linked immunosorbent assay for the quantitative detect the presence of Salbutamol in samples of urine and feed, animal’s tissue, pork’s live.
Sensitivity: 0.05 ppb
Detection Limit: Animal’s tissue and pork live 0.05 ppb / 0.2 ppb
Urine 0.05 ppb
Feed 2.5 ppb
Recovery Rate: Urine and Tissue 90% ±　15%
Pork’s live and feed 80% ±　15%
 
KIT COMPONENT

• 1 X Microtiter plate (8 microwells X 12 removable strips)
• 6 X Standard solutions (1 mL each): 0 ng/ mL, 0.05 ng/ mL, 0.15 ng/ mL, 0.45 ng/ mL, 1.35 ng/ mL, 4.05 ng/ mL
• 1 X Salbutamol antibody (6 mL)
• 1 X HRP conjugated antibody (6 mL)
• 1 X Sample dilution buffer (30 mL)
• 1 X Washing buffer concentrate (10 X , 50 mL)
• 1 X Substrate A (6 mL)
• 1 X Substrate B (6 mL)
• 1 X Stopping solution (6 mL)
• 1 X Product Manual

1. MATERIALS REQUIRED BUT NOT PROVIDED

• ELISA Microtiter plate reader equipped with 450 nm filters

- 50 – 200 μL multichannel micropipette

- 50, 100 and 200 μL precision micropipette

- Microplate washer or squeeze bottle

- Homogenizer

- Centrifuge

- Centrifugal tubes

- Distilled water

- HCl, NaOH
 
PREPARATION OF WORKING SOLUTIONS
Washing buffer: dilute 10X concentrate with distilled water (i.e. 1 mL + 9 mL).
Extract solution: mix 4% (m/v) NaCl Solution with 0.1 M HCl at the volumetric ratio 4:1.
Standard solution, Enzyme-linked conjugate, substrate A, substrate B and stopping solution are ready-to-use.
 
SAMPLE PREPARATION
 
Animal’s tissue and pork live
Method 1 (dilution factor: 4):

• Homogenize the sample (free of fat) for 1 min at 10,000 r/min.
• Weigh out 1.0 g of homogenate into a 15 mL centrifugal tube. Add 3 mL of extract solution and mix well and shake for 5 mins.
• Do a centrifugation at 5000 r/min at 20-25 °C for 5 mins.
• Take 1 mL of supernatant and mix it with 15μL of 1 M NaOH into polystyrene centrifugal tube. Adjust the pH value to 7.0-8.0.
• Take 50 μL liquid from water phase for assay.

Method 2 (dilution factor: 1):

• Homogenize the sample (free of fat) for 1 min at 10,000 r/min.
• Weigh out 2.0 g of homogenate into a 50 mL centrifugal tube. Add 6 mL of Methyl and shake for 5 mins.
• Do a centrifugation at 5000 r/min for 5 mins.
• Take 1.5 mL of supernatant (organic layer) into a glass tube and dry it by 50 °C of Nitrogen flow.
• Dissolve the residue by 0.5 mL of distilled water and add 0.5 mL of Hexane. Shake to mix well.
• Get rid of the upper layer and take 50 μL liquid for assay.
• 2.Urine (dilution factor 1)
• Take 50 μL of clear urine sample directly for assay(Do centrifuge if urine is not clear). Make dilution by distilled water before assay if the concentration of Salbutamol is high.
• 3.Feed (dilution factor 50)
• Weigh out 2 g of comminuted feed sample into a 50 mL centrifugal tube. Add 2 mL of 1 M HCl and 16 mL of distilled water. Shake to mix well.
• Do a centrifugation at 5000 r/min for 5 mins.
• Take 200 μL of supernatant and mix well with 800 μL of sample dilution buffer. Shake to mix well.
• Adjust the pH value to 7.0-8.0 by 1 M NaOH.
• Take 50 μL for assay.