IL-1 beta ELISA Kit

Catalog No. ABIN6956974

Product Details IL-1 beta ELISA Kit

Target: IL-1 beta (IL1B) (Interleukin 1, beta (IL1B))

Reactivity: Rabbit

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 7.8 pg/mL - 500 pg/mL

Minimum Detection Limit: 7.8 pg/mL

Application: ELISA

Purpose: The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of IL1b in rabbit serum, plasma, tissue homogenates, cell lysates, cell culture supernates.

Sample Type: Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate

Analytical Method: Quantitative

Specificity: This assay has high sensitivity and excellent specificity for detection of Interleukin 1 Beta (IL1b)

Sensitivity: 2.8 pg/mL

Components:

• Pre-coated, ready to use 96-well strip plate, flat buttom
• Plate sealer for 96 wells
• Reference Standard
• Standard Diluent
• Detection Reagent A
• Detection Reagent B
• Assay Diluent A
• Assay Diluent B
• Reagent Diluent (if Detection Reagent is lyophilized)
• TMB Substrate
• Stop Solution
• Wash Buffer (30 x concentrate)
• Instruction manual

Application Details

Comment:

Information on standard material:
The standard might be recombinant protein or natural protein, that will depend on the specific kit. Moreover, the expression system is E.coli or yeast or mammal cell. There is 0.05% proclin 300 in the standard as preservative.

Information on reagents:
The stop solution used in the kit is sulfuric acid with concentration of 1 mol/L. And the wash solution is TBS. The standard diluent contains 0.02 % sodium azide, assay diluent A and assay diluent B contain 0.01% sodium azide. Some kits can contain is BSA in them.

Information on antibodies:
The provided antibodies and their host vary in different kits.

Sample Volume: 100 μL

Assay Time: 3 h

Plate: Pre-coated

Protocol:

1. Prepare all reagents, samples and standards,
2. Add 100μL standard or sample to each well. Incubate 1 hours at 37 °C,
3. Aspirate and add 100μL prepared Detection Reagent A. Incubate 1 hour at 37 °C,
4. Aspirate and wash 3 times,
5. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37 °C,
6. Aspirate and wash 5 times,
7. Add 90μL Substrate Solution. Incubate 10-20 minutes at 37 °C,
8. Add 50μL Stop Solution. Read at 450nm immediately.

Reagent Preparation:

1. Bring all kit components and samples to room temperature (18-25 °C) before use. If the kit will not be used up in one time, please only take out strips and reagents for present experiment, and leave the remaining strips and reagents in required condition.
2. Standard - Reconstitute the Standard with 1.0 mL of Standard Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). The concentration of the standard in the stock solution is 10,000pg/mL. Firstly dilute the stock solution to 500pg/mL and the diluted standard serves as the highest standard (500pg/mL). Then prepare 7 tubes containing 0.5 mL Standard Diluent and use the diluted standard to produce a double dilution series. Mix each tube thoroughly before the next transfer. Set up 7 points of diluted standard such as 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.2pg/mL, 15.6pg/mL, 7.8pg/mL, and the last microcentrifuge tube with Standard Diluent is the blank as 0pg/mL.
3. Detection Reagent A and Detection Reagent B - If lyophilized reconstitute the Detection Reagent A with 150μL of Reagent Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). Briefly spin or centrifuge the stock Detection A and Detection B before use. Dilute them to the working concentration 100-fold with Assay Diluent A and B, respectively.
4. Wash Solution - Dilute 20 mL of Wash Solution concentrate (30x) with 580 mL of deionized or distilled water to prepare 600 mL of Wash Solution (1x).
5. TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.

Note:

1. Making serial dilution in the wells directly is not permitted.
2. Prepare standards within 15 minutes before assay. Please do not dissolve the reagents at 37 °C directly.
3. Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction, and avoid foaming and mix gently until the crystals are completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10μL for one pipetting.
4. The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
5. If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
6. Contaminated water or container for reagent preparation will influence the detection result.

Sample Preparation:

• It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturationmay occur in these samples, leading to false results. Samples should therefore be stored for a short periodat 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thawcycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged toremove precipitates.
• If the sample type is not specified in the instructions, a preliminary test is necessary to determinecompatibility with the kit.
• If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibilityof causing a deviation due to the introduced chemical substance.The recommended dilution factor is for reference only.
• Please estimate the concentration of the samples before performing the test. If the values are not in therange of the standard curve, the optimal sample dilution for the particular experiment has to be determined.Samples should then be diluted with PBS (pH =7.0-7.2).

Assay Precision: Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level of target were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level of target were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV < 10%
Inter-Assay: CV < 12%

Restrictions: For Research Use only

Handling

Precaution of Use: The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.

Storage: 4 °C/-20 °C

Storage Comment:

1. For unopened kit: All reagents should be stored according to the labels on the vials. The Standard, Detection Reagent A, Detection Reagent B, and 96-well Strip Plate should be stored at -20 °C upon receipt, while the other reagents should be stored at 4 °C.
2. For opened kits: the remaining reagents must be stored according to the above storage conditions. In addition, please return the unused wells to the foil pouch containing the desiccant and seal the foil pouch with the zipper.

.

Expiry Date: 6 months

References for IL-1 beta ELISA Kit (ABIN6956974)

Mikolka, Kopincova, Kosutova, Kolomaznik, Calkovska, Mokra: "Anti-IL-8 antibody potentiates the effect of exogenous surfactant in respiratory failure caused by meconium aspiration." in: Experimental lung research, Vol. 44, Issue 1, pp. 40-50, (2019) (PubMed).

Kopincova, Mikolka, Kolomaznik, Kosutova, Calkovska, Mokra: "Selective inhibition of NF-kappaB and surfactant therapy in experimental meconium-induced lung injury." in: Physiological research, Vol. 66, Issue Suppl 2, pp. S227-S236, (2018) (PubMed).

Lin, Xiao, Liu, Chen, Zhu, Gao, Gao, Tong, Zhang, Li, Lin, Li, Liang, Lin, Liu, Yang: "Development of tissue inflammation accompanied by NLRP3 inflammasome activation in rabbits infected with Treponema pallidum strain Nichols." in: BMC infectious diseases, Vol. 18, Issue 1, pp. 101, (2018) (PubMed).

Ma, Zhu, Chen, Zhang, Dong, Wu, Lu, Wu: "Sulforaphane, a Natural Isothiocyanate Compound, Improves Cardiac Function and Remodeling by Inhibiting Oxidative Stress and Inflammation in a Rabbit Model of Chronic Heart Failure." in: Medical science monitor : international medical journal of experimental and clinical research, Vol. 24, pp. 1473-1483, (2018) (PubMed).

Kopincova, Mikolka, Kolomaznik, Kosutova, Calkovska, Mokra: "Modified porcine surfactant enriched by recombinant human superoxide dismutase for experimental meconium aspiration syndrome." in: Life sciences, Vol. 203, pp. 121-128, (2018) (PubMed).

Rambaud, Lidouren, Sage, Kohlhauer, Nadeau, Fortin-Pellerin, Micheau, Zilberstein, Mongardon, Ricard, Terada, Bruneval, Berdeaux, Ghaleh, Walti, Tissier: "Hypothermic total liquid ventilation after experimental aspiration-associated acute respiratory distress syndrome." in: Annals of intensive care, Vol. 8, Issue 1, pp. 57, (2018) (PubMed).

Mokra, Kosutova, Balentova, Adamkov, Mikolka, Mokry, Antosova, Calkovska: "Effects of budesonide on the lung functions, inflammation and apoptosis in a saline-lavage model of acute lung injury." in: Journal of physiology and pharmacology : an official journal of the Polish Physiological Society, Vol. 67, Issue 6, pp. 919-932, (2017) (PubMed).

Crane, Broome, Lis: "Biological Activities of Uric Acid in Infection Due to Enteropathogenic and Shiga-Toxigenic Escherichia coli." in: Infection and immunity, Vol. 84, Issue 4, pp. 976-88, (2016) (PubMed).

Liu, Peng, Fang, Yao, Redmond, Ni: "No midterm advantages in the middle term using small intestinal submucosa and human amniotic membrane in Achilles tendon transverse tenotomy." in: Journal of orthopaedic surgery and research, Vol. 11, Issue 1, pp. 125, (2016) (PubMed).

Kosutova, Mikolka, Balentova, Adamkov, Kolomaznik, Calkovska, Mokra: "Intravenous dexamethasone attenuated inflammation and influenced apoptosis of lung cells in an experimental model of acute lung injury." in: Physiological research, Vol. 65, Issue Supplementum 5, pp. S663-S672, (2016) (PubMed).

Yu, Xie, Xin, Wang: "Panax notoginseng saponins promote wound repair of anterior cruciate ligament through phosphorylation of PI3K, AKT and ERK." in: International journal of clinical and experimental pathology, Vol. 8, Issue 1, pp. 441-9, (2015) (PubMed).

Joos, Leucht, Riegger, Hogrefe, Fiedler, Dürselen, Reichel, Ignatius, Brenner: "Differential Interactive Effects of Cartilage Traumatization and Blood Exposure In Vitro and In Vivo." in: The American journal of sports medicine, Vol. 43, Issue 11, pp. 2822-32, (2015) (PubMed).

Ortencio, Renzo, Sobrinho B, Kobashigawa, Crivelaro, Silva, Aldrovani, Ribeiro, Mineo, Laus: "Effects of morphine on the expression of cytokines and inflammatory mediators in a rabbit model of endotoxin-induced experimental uveitis." in: Arquivos brasileiros de oftalmologia, Vol. 78, Issue 6, pp. 371-5, (2015) (PubMed).

Li, Liu, Xiang, Wang, Lee, Zhang: "Therapeutic potential of the antimicrobial peptide OH-CATH30 for antibiotic-resistant Pseudomonas aeruginosa keratitis." in: Antimicrobial agents and chemotherapy, Vol. 58, Issue 6, pp. 3144-50, (2014) (PubMed).

Chen, Lin, You, Lei, Li, Lin, Luo, Yan: "Hyperlipidemia causes changes in inflammatory responses to periodontal pathogen challenge: implications in acute and chronic infections." in: Archives of oral biology, Vol. 59, Issue 10, pp. 1075-84, (2014) (PubMed).

Kim, Ko, Ko, Lee, Kim, Wee, Lee, Fulcher, Oh: "TSG-6 protects corneal endothelium from transcorneal cryoinjury in rabbits." in: Investigative ophthalmology & visual science, Vol. 55, Issue 8, pp. 4905-12, (2014) (PubMed).

Miroshnikov, Reznikov, Kokhanov: "[Comparative evaluation of the action of serum proteins on tissue regeneration]." in: Biulleten' eksperimental'no? biologii i meditsiny, Vol. 103, Issue 1, pp. 95-7, (1987) (PubMed).

Target Details for IL-1 beta

Target: IL-1 beta (IL1B) (Interleukin 1, beta (IL1B))

Alternative Name: Interleukin 1 Beta (IL1b) (IL1B Products)

Synonyms: IL-1 ELISA Kit, IL1-BETA ELISA Kit, IL1F2 ELISA Kit, IL-1BETA ELISA Kit, IL1beta ELISA Kit, il1-b ELISA Kit, zgc:111873 ELISA Kit, IL-1B ELISA Kit, IL-1beta ELISA Kit, Il-1b ELISA Kit, IL1B ELISA Kit, IL-1 beta ELISA Kit, IL-1b ELISA Kit, interleukin 1 beta ELISA Kit, interleukin 1, beta ELISA Kit, IL1B ELISA Kit, il1b ELISA Kit, Il1b ELISA Kit

Pathways: NF-kappaB Signaling, Interferon-gamma Pathway, TLR Signaling, Negative Regulation of Hormone Secretion, Cellular Response to Molecule of Bacterial Origin, Carbohydrate Homeostasis, Glycosaminoglycan Metabolic Process, Myometrial Relaxation and Contraction, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Autophagy, Cancer Immune Checkpoints, Inflammasome