IL-8 ELISA Kit

Produktnummer ABIN1115681

Dieses Colorimetric ELISA kit dient der quantitativen Messung von Human IL-8. Es wurde in 4 Publikationen zitiert.

Kurzübersicht für IL-8 ELISA Kit (ABIN1115681)

Target: IL-8 (IL8) (Interleukin 8 (IL8))

Reaktivität: Human

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 7.81 pg/mL - 500 pg/mL

Applikation: ELISA

Proben: Plasma, Serum

Produktdetails IL-8 ELISA Kit

Untere Nachweisgrenze: 7.81 pg/mL

Verwendungszweck: The kit is a sandwich enzyme immunoassay technique for the in vitro quantitative measurement in various sample types.

Analytische Methode: Quantitative

Spezifität: This assay has high sensitivity and excellent specificity for detection of the antigen.

Kreuzreaktivität (Details): No significant cross-reactivity or interference between human Interleukin 8 and analogues was observed. Note: Limited by current skills and knowledge, it is impossible for us to complete the cross-reactivity detection between Human bFGF and all the analogues, therefore, cross reaction may still exist.

Sensitivität: 4.69 pg/mL

Bestandteile:

• Pre-coated, ready to use 96-well strip plate, flat buttom
• Plate sealer for 96 wells
• Reference Standard
• Reference Standard & Sample Diluent
• Biotinylated Detection Antibody (100 x concentrate)
• HRP Conjugate (100 x concentrate)
• Biotinylated Detection Antibody Diluent
• HRP Conjugate Diluent
• Substrate Reagent
• Stop Solution
• Wash Buffer (25 x concentrate)
• Instruction manual

Anwendungsinformationen

Applikationshinweise: ELISA Plate: The just opened ELISA Plate may appear water-like substance, which is normal and will not have any impact on the experiment results.
Add Sample: The interval of sample adding between the first well and the last well should not be too long, otherwise will cause different pre-incubation time, which will significantly affect the experiment’s accuracy and repeatability. For each step in the procedure, total dispensing time for addition of reagents or samples to the assay plate should not exceed 10 minutes. Parallel measure ment is recommended.
Incubation: To prevent evaporation and ensure accurate results, proper adhesion of plate sealers during incubation steps is necessary. Do not allow wells to sit uncovered for extended periods between incubation steps. Do not let the strips dry at any time during the assay. Strict compliance with the given incubation time and temperature.
Washing: The wash procedure is critical. Insufficient washing will result in poor precision and falsely elevated absorbance readings. Residual liquid in the reaction wells should be pat dry against absorbent paper in the washing process. But don’t put absorbent paper into reaction wells directly. Note that clear the residual liquid and fingerprint in the bottom before measurement, so as not to affect the micro-titer plate reader.
Reagent Preparation: As the volume of Detection Ab and HRP Conjugate is very small, liquid may adhere to the tube wall or tube cap when being transported. You better hand-throw it or centrifugal it for 1 minute at 1000rpm. Please pipette the solution for 4-5 times before pippeting. Please carefully reconstitute Standards, working solutions of Detection Ab and HRP Conjugate according to the instructions. To minimize imprecision caused by pipetting, ensure that pipettors are calibrated. It is recommended to suck more than 10μL for once pipetting. Do not reuse standard solution, working solution of Detection Ab and HRP Conjugate, which have been diluted. If you need to use standard repeatedly, you can divide the standard into small pack according to the amount of each assay, keep them at -20°C to -80°C and avoid repeated freezing and thawing.
Reaction Time Control: Please control reaction time strictly following this product description!
Substrate: Substrate Solution is easily contaminated. Please protect it from light.Stop Solution: As it is an acid solution, please pay attention to the protection of your eyes, hands, face and clothes when using this solution.
Mixing: You’d better use micro-oscillator at the lowest frequency, as sufficient and gentle mixing is particularly important to reaction result. If there is no micro-oscillator available, you can knock the ELISA plate frame gently with your finger before reaction.
Security: Please wear lab coats and latex gloves for protection. Especially detecting samples of blood or other body fluid, please perform following the national security columns of biological laboratories.
Do not use component from different batches of kit(washing buffer and stop solution can be an exception)
To avoid cross-contamination, change pipette tips between adding of each standard level, between sample adding, and between reagent adding. Also, use separate reservoirs for each reagent. Otherwise, the results will be inaccurate!

Kommentare:

Information on standard material:
The formulation of the standard is 0.01 M PBS. The standard contains additives (1 % BSA).

Information on reagents:
Reagents include 1 M SO₂. Azide, thimerosal, 2-mercaptoethanol (2-ME) or any other poisonous materials are not used.

Information on antibodies:
The provided antibodies and their host vary in different kits. All antibodies are affinity purified

The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest protein concentration that could be differentiated from zero. It was determined by adding two standard deviations to the mean optical density value of twenty zero standard replicates and calculating the corresponding concentration.

Probenmenge: 100 µL

Plattentyp: Pre-coated

Protokoll:

1. Add 100 µL standard or sample to each well. Incubate for 90 min at 37 °C.
2. Remove the liquid. Add 100 µL Biotinylated Detection Antibody. Incubate for 1 hour at 37 °C.
3. Aspirate and wash 3 times.
4. Add 100 µL HRP Conjugate. Incubate for 30 min at 37 °C.
5. Aspirate and wash 5 times.
6. Add 90 µL Substrate Reagent. Incubate for 15 min at 37 °C.
7. Add 50 µL Stop Solution. Read at 450 nm immediately.
8. Calculation of results.

Aufbereitung der Reagenzien:

1. Equilibrate all reagents to room temperature (18-25 °C) before use. If the kit will not be used up in one assay, please only take out the necessary strips and reagents for present experiment, and store the remaining strips and reagents at required condition.
2. Standard Working Solution: Centrifuge the standard at 10,000x g for 1 min. Reconstitute with 1 mL of the Standard & Sample Diluent, allow to stand for 10 minutes, and gently invert several times until completely dissolved. Alternatively, vortex briefly at low speed, remove bubbles by low-speed centrifugation if necessary.This produces a 500 pg/mL stock solution. Prepare serial dilutions according to experimental requirements. Recommended dilution gradient: 500, 250, 125, 62.5, 31.25, 15.63, 7.81, 0 pg/mL Serial Dilution Procedure: Dispense 500 μL of Standard & Sample Diluent into 7 EP tubes, respectively. Transfer 500 μL of the stock solution into the first tube. Sequentially transfer 500 μL from the preceding tube into the next tube for subsequent dilutions. Designate the last tube as the blank, do not transfer any solution into it. Aliquot the stock and store at -20 ° C. Use within two weeks and avoid repeated freeze-thaw cycles. Serially diluted standards should be prepared immediately, prior to use.
3. Biotinylated Detection Ab working solution: Calculate the required volume (100 μL/well) plus a slight excess. Centrifuge the Concentrated Biotinylated Detection Antibody (100x) at 800 x g for 1 minute. Dilute to 1x with Biotinylated Detection Ab Diluent at a ratio of 1:99. Prepare the working solution immediately, prior to use.
4. HRP Conjugate Working Solution: Calculate the required volume (100 μL/well) plus a slight excess. Centrifuge the Concentrated HRP Conjugate (100x) at 800 x g for 1 minute. Centrifuge the Concentrated HRP Conjugate (100x) at 800 x g for 1 minute. Prepare the working solution immediately, prior to use.
5. Wash Buffer: Prepare the working Wash Buffer by diluting 30 mL of Concentrated Wash Buffer with 720 mL of deionized or distilled water yielding 750 mL. Note: if crystals have formed in the concentrate, warm it in a 40 °C water bath and mix it gently until the crystals have completely dissolved. For same day use only.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Handhabung: All the reagents in the kit should be stored according to the labels on vials. Unused wells should be returned to the foil pouch with the desiccant pack and resealed along entire edge of zip-seal. Substrate Reagent shouldn't be kept at -20 °C (Check!). Exposure of reagents to strong light should be avoided in the process of incubation and storage. All the taps of reagents should be tightened to prevent evaporation and microbial contamination. If not to store reagents according to above suggestions, erroneous results may occur.

Lagerung: 4 °C/-20 °C

Informationen zur Lagerung: The unopened kit can be stored at 4℃ for 1 month. If the kit is not used within 1 month, store the items separately according to the conditions since the kit is received.

Referenzen für IL-8 ELISA Kit (ABIN1115681)

Ruan, Xu, Wang, Wu, Zhu, Ren, Bian, Chang, Bai, Han, Ding: "Associations between knee structural measures, circulating inflammatory factors and MMP13 in patients with knee osteoarthritis." in: Osteoarthritis and cartilage, Vol. 26, Issue 8, pp. 1063-1069, (2018) (PubMed).

Tian, Li, He, Song, Hou, Tian, Meng, Li, Shan: "Antidiabetic (type 2) effects of Lactobacillus G15 and Q14 in rats through regulation of intestinal permeability and microbiota." in: Food & function, Vol. 7, Issue 9, pp. 3789-3797, (2018) (PubMed).

Liu, Wu, Song, Liang, Zheng, Xiao, Liu, Zouboulis, Lei: "Particulate matter 2.5 regulates lipid synthesis and inflammatory cytokine production in human SZ95 sebocytes." in: International journal of molecular medicine, Vol. 40, Issue 4, pp. 1029-1036, (2018) (PubMed).

Yan, Wang, Wu, Wang, Li, Liang, Wu: "Zinc oxide nanoparticle-induced atherosclerotic alterations in vitro and in vivo." in: International journal of nanomedicine, Vol. 12, pp. 4433-4442, (2017) (PubMed).

Details zu IL-8

Target: IL-8 (IL8) (Interleukin 8 (IL8))

Andere Bezeichnung: Interleukin 8

Hintergrund: Synonyms: IL8, CXCL8, GCP-1, GCP1, LECT, LUCT, LYNAP, MDNCF, MONAP, NAF, NAP-1, NAP1, 3-10C, AMCF-I, K60

UniProt: P10145

Pathways: TLR Signalweg, Cellular Response to Molecule of Bacterial Origin, Regulation of G-Protein Coupled Receptor Protein Signaling, ER-Nucleus Signaling, Hepatitis C, Autophagie