IL1A Kit ELISA
Aperçu rapide pour IL1A Kit ELISA (ABIN7671644)
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Méthode de détection
Type de méthode
Gamme de detection
Application
Type d'échantillon
Classe de qualité
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Seuil minimal de détection
- 3.906 pg/mL
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Fonction
- Human IL-1α (Interleukin 1 Alpha) QuickTest ELISA Kit
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Analytical Method
- Quantitative
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Specificité
- Specifically binds with IL-1α , no obvious cross reaction with other analogues. Recommended Sample Dilution Ratio The following table shows the recommended dilution ratios for this kit for a limited number of samples for your reference only. (When the sample content is very low, the original solution can be added without dilution, but it is necessary to ensure that the pH is between 6.8 and 8.0, and it does not contain more than 10 % organic solvents or high-concentration protein denaturants.) 1. Common sample validation: Sample Type Recommended Dilution Ratio Content Healthy serum (n=18) undiluted ND-4.6pg/mL Healthy plasma (EDTA, Citrate , heparin) (n=16) undiluted ND-4.1pg/mL Human peripheral blood cells were cultured in RPMI 1640 supplemented with 10 %FBS, 100 units/mL penicillin, and 100 μg/mL streptomycin sulfate, followed by stimulated with 10 μg/mL PHA for 24 hours. The cell culture supernatants were assayed for levels of human IL-1a. 1/2-1/5 dilution 725pg/mL THP-1 cells were treated with 80nM TPA and cultured overnight.The cell culture supernatants were assayed for levels of human IL-1a. undiluted ND THP-1 cells were treated with 80nM TPA and cultured overnight. Then, stimulate with 100 ng/mL LPS for 6 hours and take the cell culture supernatant were assayed for levels of human IL-1a. undiluted 196pg/mL 2. KD sample validation (Detect IL-1α-KD THP-1 cells): Sample Type Dilution Ratio Content Wild THP-1 cells were treated with 80nM TPA and cultured overnight, and then stimulated with 200 ng/mL LPS for 6 hours. The cell culture supernatants were assayed for levels of human IL-1a. undiluted 206pg/mL THP-1 cells (+siRNA) were treated with 80nM TPA and cultured overnight, and then stimulated with 200 ng/mL LPS for 6 hours. The cell culture supernatants were assayed for levels of human IL-1a. undiluted 21pg/mL Note:ND is lower than the sensitivity of the kit and was not detected 3. Verified recombinant protein: (It is a normal phenomenon that some proteins have weak detection signals or cannot be detected at all due to differences in tags, sequences or protein activities) A. HEK293-derived Human IL-1 alpha protein Ser113-Ala271B. E. coli-derived Human IL-1 alpha protein Ser113-Ala271
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Sensibilité
- 2.344 pg/mL
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Ingrédients
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- ELISA Microplate(Dismountable)
- Lyophilized Standard
- Cap/Det Ab (Ready to use
- blue)
- TMB Substrate
- Sample Dilution Buffer
- Stop Solution
- Plate Sealer
- Product Description
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Matériel non inclus
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- Microplate reader (wavelength: 450nm)
- 37 °C incubator (CO2 incubator for cell culture is not recommenced.)
- Automated plate washer or multi-channel pipette/5 mL pipettor
- Precision single (0.5-10μL
- 5-50μL
- 20-200μL
- 200-1000μL) and multi-channel pipette with disposable tips(Calibration is required before use.)
- Sterile tubes and Eppendorf tubes with disposable tips
- Absorbent paper and loading slot
- Deionized or distilled water
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Indications d'application
- Optimal working dilution should be determined by the investigator.
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Commentaires
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Sandwich ELISA, Double Antibody
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Volume d'échantillon
- 50 μL
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Durée du test
- 2 h
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Plaque
- Pre-coated
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Protocole
- Step 1: Take out the required plate wells, add 50 µL Cap/Det Ab into each well, then add 50 µL Standard or Sample into individual well. (When adding standard or sample, the disposable tip lightly touches the liquid level. Change the disposable tips for different samples and standards.) Gently tap the plate for 10s to ensure thorough mixing then static incubate for 60 minutes at 37 °C. Washing: Wash the plate twice without immersion. Step 2: Add 100 µL HRP-Streptavidin (orange) into each well, seal the plate and static incubate for 30 minutes at 37 °C. Washing: Wash the plate five times without immersion. Step 3: Add 90 µL TMB substrate solution, seal the plate and static incubate for 10-20 minutes at 37 °C. (Accurate TMB visualization control is required.) Step 4: Add 50 µL stop solution. Read at 450nm immediately and calculate.
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Procédure de l'essai
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- Take out the required strips, add 50 µL Cap/Det Ab into each well, then add 50 µL Standard or Sample into relevant well. (When adding standard or sample, the disposable tip lightly touches the liquid level. Change the disposable tips for different samples and standards.) Gently tap the plate for 10s to ensure complete mixing then statically incubate for 60 minutes at 37 °C.
- Wash the plate twice without immersion.
- Add 100 µL HRP-Streptavidin (orange) into each well, seal the plate and statically incubate for 30 minutes at 37 °C.
- Wash the plate five times without immersion.
- Add 90 µL TMB substrate solution, seal the plate and statically incubate for 10-20 minutes at 37 °C (Accurate TMB visualization control is required.).
- Add 50 µL stop solution. Read at 450nm immediately and calculate.
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Restrictions
- For Research Use only
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Conseil sur la manipulation
- Avoid direct light exposure to antibody solutions
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Stock
- 4 °C
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Stockage commentaire
- Store sealed Kit at 2-8° C. Do Not Cryopreserve. Store ELISA Microplate for 1 month at 2-8°C, Store for 12 month at -20°C. Put the leftover strips into a sealed foil bag with the desiccant
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Date de péremption
- 12 months
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- IL1A (Interleukin 1 alpha (IL1A))
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Autre désignation
- IL-1alpha
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Sujet
- Interleukin-1 alpha, IL-1 alpha, Hematopoietin-1, IL1A, IL1F1
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UniProt
- P01583
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Pathways
- Signalisation NF-kappaB, Autophagy, Cancer Immune Checkpoints
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