Products
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EIS25458Mo
Simian IL4 ELISpot Kit
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Alias：BSF1, IL-4, BCGF1, BSF-1, BCGF-1
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Specification：96-well PVDF-backed microplate.
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Storage Conditions：-20℃; short-term storage at 4℃ is acceptable for up to 2 weeks.
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Application：ELISpot
Download The Manual
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Kit Components
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Name
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96 Tests
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48 Tests
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Storage
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Pre-coated plate
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8×12
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8×6
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4/-20℃
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Positive Control
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2 vial
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1 vial
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4/-20℃
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Biotinylated antibody (1:100)
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1 vial
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1 vial
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4/-20℃
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Enzyme conjugate(1:100)
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1 vial
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1 vial
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4/-20℃
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Antibody diluent
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1 vial
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1 vial
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4/-20℃
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Enzyme diluent
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1 vial
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1 vial
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4/-20℃
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Washing buffer (1:25)
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1 vial
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1 vial
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4/-20℃
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Color Reagent(BCIP/NBT Substrate)
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1 vial
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1 vial
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4/-20℃
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Attention
Color Reagent: Store protected from light.
Self-Prepared Equipment
- Dissection microscope or an ELISpot reader.
- Pipettes and pipette tips.
- Deionized water.
- Squirt bottle, manifold dispenser, or automated microplate washer.
- 500 mL graduated cylinder.
- 37 °C CO2 incubator.
- Sterile culture media.
Other Self-Provided Materials
- Centrifuge tubes (capacities: 1.5 mL, 5 mL, etc.)
- Disposable pipette tips (ranges: 0.5–10 μL, 20–200 μL, 200–1000 μL)
- Pure water or distilled water
Sample Collection and Preparation
Sample Preparation
01.The types of effector and responder cells used, method of cell separation, mode of stimulation, and length of incubation are to be determined by each investigator.
02.Avoid cell contamination throughout the procedure.
Experimental Procedure
Experimental Procedure
Result Calculation
The developed microplate can be analyzed by counting spots using either a dissection microscope or an ELISpot reader. Specific spots are round and have a dark center with slightly fuzzy edges. Quantitation of results can be done.
Performance Metrics
Precautions
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01The reconstituted positive control cannot be stored after preparation; do not refreeze the positive control once it has been reconstituted.
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02Due to shaking or inversion during transportation, centrifugation of the kit tubes or bottles may be necessary to collect all contents at the bottom. Tubes should be vortexed manually or centrifuged at 1000 rpm for 1 min.
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03Concentrated washing buffer may crystallize slightly. Heat the buffer in a water bath to facilitate dissolution during dilution. Crystals must be completely dissolved before the washing buffer is used.
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04The prepared positive control is for single-use only; do not reuse any positive control that has already been assayed. Use the second vial provided if the assay needs to be repeated.
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05Use only the reagents and components provided with this kit. Do not mix batches or lots from other orders of this kit or from different kit types.
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06Ensure all reagents are thoroughly mixed. For reagents added to the microplate, adequate mixing is critical for obtaining accurate assay results.
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07Bring the kit to room temperature before starting the assay.
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08Store unused microplate strips in the original foil bag at 2–8℃ if they will be used within a short period.
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09Do not use kits that have exceeded their expiration date.
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10All samples, washing buffer, and wastes should be treated as biohazardous waste.
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11Sample addition must be performed using a pipette or similar calibrated instrument. Calibrate the instrument before the assay to avoid experimental errors. Add samples to the wells quickly, and control the total sample addition time to less than 5 minutes. A multichannel pipette is recommended to reduce loading time.
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12Regularly verify the calibration of the incubator to ensure a stable incubation temperature of 37℃.
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13For the 48-well ELISpot Kit, all component volumes are 50% of those provided in the 96-well kit.