Mouse IL-1β(Interleukin 1 Beta) ELISA Kit

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This kit is based on Double antibody-Sandwich ELISA detection method and takes 4h assay time. The microplate provided in this kit has been precoated with anti IL-1β antibody. Add standard and properly diluted sample into relevant well respectively. After incubation, wash unbound components. Add biotinylated detection antibody. Then, it binds with IL-1β bound to precoated antibody. Wash unbound components and add HRP-Streptavidin Conjugate (SABC). Wash unbound components again and add TMB substrate solution. Then, TMB was catalyzed by HRP to produce a blue color product that turned yellow after adding a stop solution. Read the O.D. absorbance at 450nm in a microplate reader. Calculate the concentration of IL-1β in the sample by plotting standard curve. The concentration of the target substance is proportional to the OD450 value.

Catalogue No.: 
EM0109
Alias: 
Interleukin-1 beta ELISA Kit, IL-1 beta ELISA Kit, Catabolin ELISA Kit, IL1B ELISA Kit, IL1F2 ELISA Kit
Species: 
Mouse
Range: 
12.5-800pg/ml (1pg=1100mIU)
Sensitivity: 
7.5pg/ml
Size Price
96T Inquiry
48T Inquiry
Dispatch Time: About 3 working days
  • SPECIFICATIONS
  • CITATIONS
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  • CONDITIONS
  • FAQS
Product Name
Mouse IL-1β(Interleukin 1 Beta) ELISA Kit
Alias
Interleukin-1 beta ELISA Kit, IL-1 beta ELISA Kit, Catabolin ELISA Kit, IL1B ELISA Kit, IL1F2 ELISA Kit
Catalogue No.
EM0109
Size
48T/96T
Species
Mouse
UniProt ID
P10749
Sample Type
Serum, Plasma, Cell Culture Supernatant, cell or tissue lysate, Other liquid samples
Detection Method
Sandwich ELISA, Double Antibody
Detection Wavelength
OD450
Reaction Duration
4 hours
Range
12.5-800pg/ml (1pg=1100mIU)
Sensitivity
7.5pg/ml
Storage
2-8°C(Sealed), Don't cryopreserve.
Specificity
Specifically binds with IL-1β , no obvious cross reaction with other analogues.
ELISA Kit Components
Kit Components Item Size(48T) Size(96T) Storage Condition for Opened Kit
E001 ELISA Microplate(Dismountable) 8×6 8×12 Put the rest strips into a sealed foil bag with the desiccant. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C
E002 Lyophilized Standard 1vial 2vial Put the rest standards into a desiccant bag. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C
E003 Biotin-labeled Antibody(Concentrated, 100X) 60ul 120ul 2-8°C (Avoid Direct Light)
E034 HRP-Streptavidin Conjugate(SABC, 100X) 60ul 120ul
E024 TMB Substrate 5ml 10ml
E039 Sample Dilution Buffer 10ml 20ml 2-8°C
E040 Antibody Dilution Buffer 5ml 10ml
E049 SABC Dilution Buffer 5ml 10ml
E026 Stop Solution 5ml 10ml
E038 Wash Buffer(Concentrated, 25X) 15ml 30ml
E006 Plate Sealer 3 pieces 5 pieces  
E007 Product Description 1 copy 1 copy
Required Instruments and Reagents
  1. Microplate reader (wavelength: 450nm)
  2. 37°C incubator (CO2 incubator for cell culture is not recommenced.)
  3. Automated plate washer or multi-channel pipette/5ml pipettor (for manual washing purpose)
  4. Precision single (0.5-10μL, 5-50μL, 20-200μL, 200-1000μL) and multi-channel pipette with disposable tips(Calibration is required before use.)
  5. Sterile tubes and Eppendorf tubes with disposable tips
  6. Absorbent paper and loading slot
  7. Deionized or distilled water
Assay Procedure Summary
Elisa实验原理图
  • Step 1: Add 100ul standard or sample into each well, seal the plate and statically incubate for 90 minutes at 37°C.
  • Washing: Wash the plate twice without immersing.
  • Step 2: Add 100ul biotin-antibody working solution, seal the plate and statically incubate for 60 minutes at 37°C.
  • Washing: Wash the plate three times and immerse for 1min each time.
  • Step 3: Add 100ul HRP-Streptavidin Conjugate (SABC) working solution, seal the plate and statically incubate for 30 minutes at 37°C.
  • Washing: Wash the plate five times and immerse for 1min each time.
  • Step 4: Add 90ul TMB substrate solution, seal the plate and statically incubate for 10-20 minutes at 37°C. (Accurate TMB visualization control is required.)
  • Step 5: Add 50ul stop solution. Read at 450nm immediately and calculate.
Standard Curve

This product is detected by QC department and meets performance required in the manual. (Laboratory Humidity: 20%-60%; Temperature: 18°C -25°C; Equilibrate TMB substrate to 37°C before staining. After adding into the ELISA wells, incubate for 15min at 37°C in dark.)

Due to different assay environments and operations, assay data below and standard curve are provided for reference. Experimenters should establish standard curve according to their own assay.

STD.(pg/ml) OD-1 OD-2 Average
0 0.104 0.11 0.106
12.5 0.142 0.149 0.145
25 0.195 0.205 0.199
50 0.288 0.303 0.294
100 0.444 0.466 0.453
200 0.764 0.803 0.779
400 1.372 1.442 1.4
800 2.05 2.154 2.091
EM0109 Standard Curve Image
Recovery

Add a certain amount of IL-1β into the sample. Calculate the recovery by comparing the measured value with the expected amount of IL-1β in the sample.

Sample Type Recovery Range(%) Average(%)
serum(n=10) 96-104 99
EDTA plasma(n=10) 94-103 98
Heparin plasma(n=10) 89-96 92
Linearity

Dilute the sample with a certain amount of IL-1β at 1:2, 1:4 and 1:8 to get the recovery range.

Sample Type 1:2 1:4 1:8
serum(n=10) 85-104% 90-105% 85-103%
EDTA plasma(n=10) 84-92% 92-101% 86-101%
Heparin plasma(n=10) 81-94% 80-91% 81-99%
Precision(%)

Intra-assay Precision: samples with low, medium and high concentration are tested 20 times on the same plate.

Inter-assay Precision: samples with low, medium and high concentration are tested 20 times on three different plates.

Item Intra-assay Precision Inter-assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/ml) 25.9 104.2 399 26.2 103.1 408
Standard deviation 1.21 4.89 20.87 1.45 4.84 22.15
CV(%) 4.69 4.69 5.23 5.54 4.69 5.43
Stability

Perform the stability test for the sealed kit at 37°C and 2-8°C and get relevant data.

ELISA kit(n=5) 37°C for 1 month 2-8°C for 6 months
Average(%) 80 95-100
IF: 17.521
Journal:
Advanced Science
Author:
The People's Hospital of Gaozhou Guangdong Medical University Maoming 525200 China
Cited Date:
2023-04-28
Product:
IF: 13.4
Journal:
Chemical Engineering Journal
Author:
School of Pharmacy, Shenyang Pharmaceutical University, No.103 Wenhua Road, Shenhe District, Shenyang 110016, China
Sample:
LO2 cells
Cited Date:
2025-03-28
Product:
IF: 10.6
Journal:
Journal of Nanobiotechnology
Author:
Integrative Science Center of Germplasm Creation in Western China (Chongqing) Science City, Biological Science Research Center, Southwest University, Chongqing, 400715, China.
Sample:
cell culture supernatants
Cited Date:
2025-01-17
Product:
IF: 10.6
Journal:
Journal of Nanobiotechnology
Author:
School of First Clinical Medical, Ningxia Medical University, Yinchuan, 750004, China.
Sample:
supernatant
Cited Date:
2025-01-24
Product:
IF: 9.1
Journal:
Pharmacological Research
Author:
State Key Laboratory of Organ Failure Research, National Clinical Research Center of Kidney Disease, Guangdong Provincial Clinical Research Center for Kidney Disease, Guangdong Provincial Key Laboratory of Nephrology, Division of Nephrology, Nanfang Hospi
Cited Date:
2024-09-06
Product:
IF: 8.2
Journal:
International Journal of Biological Macromolecules
Author:
College of Food Science and Engineering, Shanxi Agricultural University, Taigu, Shanxi 030801, China; Shanxi Key Laboratory of Edible Fungi for Loess Plateau, Taigu, Shanxi 030801, China.
Cited Date:
2024-04-07
Product:
IF: 5.8
Journal:
Biochemical Pharmacology
Author:
Shengli Clinical Medical College of Fujian Medical University, Fuzhou, Fujian, China; Fourth Department of Critical Care Medicine, Fujian Provincial Hospital, Fujian Provincial Key Laboratory of Emergency Medicine, Fuzhou, Fujian, China.
Cited Date:
2024-04-07
Product:
IF: 4.85
Journal:
Cellular Signalling
Author:
Department of Cardiology, Guangzhou Institute of Cardiovascular Disease, Guangdong Key Laboratory of Vascular Diseases, State Key Laboratory of Respiratory Disease, The Second Affiliated Hospital, Guangzhou Medical University, Guangzhou 510260, China
Cited Date:
2023-05-12
Product:
IF: 4.8
Journal:
International Immunopharmacology
Author:
Department of General Surgery, Nanfang Hospital, Southern Medical University, Guangdong Provincial Key Laboratory of Precision Medicine for Gastrointestinal Tumor, Guangzhou, Guangdong, China.
Cited Date:
2024-09-06
Product:
IF: 4.8
Journal:
Nutrients
Author:
Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Campus Zacatenco, Unidad Profesional Adolfo López Mateos, Zacatenco, Av. Wilfrido Massieu 399, Colonia Nueva Industrial Vallejo, Alcaldía Gustavo A. Madero, Ciudad de México 07738, M
Sample:
supernatant
Cited Date:
2025-02-28
Product:
IF: 4.4
Journal:
Neurochemical Research
Author:
Department of Anesthesiology, Affiliated Hospital of North Sichuan Medical College, Nanchong, China
Cited Date:
2023-09-01
Product:
IF: 4.1
Journal:
Neuroendocrinology
Author:
Department of Endocrinology, Renmin Hospital of Wuhan University, Wuchang District, Wuhan, Hubei, China
Cited Date:
2024-01-05
Product:
IF: 3.9
Journal:
Functional & Integrative Genomics
Author:
Department of Nephrology, Chongqing Traditional Chinese Medicine Hospital, No.6 Panxi Road, Jiangbei District, Chongqing, 400021, China.
Cited Date:
2024-08-02
Product:
IF: 3.8
Journal:
Acta Diabetologica
Author:
Department of Endocrinology and Metabolic Diseases, Dalian University Affiliated Xinhua Hospital, Dalian, 116021, Liaoning, China
Cited Date:
2023-10-20
Product:
IF: 3.712
Journal:
Immunopharmacology and Immunotoxicology
Author:
Department of Respiratory Medicine, Baoshan Branch of Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, China
Cited Date:
2023-07-14
Product:
IF: 3.7
Journal:
Animal Models and Experimental Medicine
Author:
Sichuan Provincial Engineering Laboratory for Prevention and Control Technology of Veterinary Drug Residue in Animal-Origin Food, Chengdu Medical College, Chengdu, PR China.
Sample:
serum
Cited Date:
2024-05-17
Product:
IF: 3.5
Journal:
Vascular Pharmacology
Author:
Department of Clinical and Biological Sciences, University of Turin, Regione Gonzole 10, Orbassano, TO 10043, Italy.
Sample:
supernatants
Cited Date:
2025-03-21
Product:
IF: 3.3
Journal:
Toxicology and Applied Pharmacology
Author:
Department of Pharmacology, Faculty of Veterinary Medicine, Mansoura University, Mansoura 35516, Egypt.
Cited Date:
2024-09-27
Product:
IF: 3
Journal:
FEBS Letters
Author:
Department of Biological Sciences, Bose Institute, Unified Academic Campus, Kolkata, India.
Sample:
supernatants
Cited Date:
2024-08-23
Product:
IF: 2.7
Journal:
Tissue and Cell
Author:
Department of Medical Analysis, Princess Aisha Bint Al-Hussein College of Nursing and Health Sciences, Al-Hussein Bin Talal University, Ma'an 71111, Jordan.
Cited Date:
2025-03-07
Product:
IF: 2.5
Journal:
Biochemical and Biophysical Research Communications
Author:
Institute of Materia Medica, School of Pharmacy, North Sichuan Medical College, Nanchong, 637000, China
Sample:
supernatant
Cited Date:
2024-12-06
Product:
IF: 1.7
Journal:
Biologicals
Author:
Department of Biotechnology, Natural and Applied Sciences Institute, Ege University, 35100, ?zmir, Turkey
Cited Date:
2023-12-22
Product:
Journal:
Medical Journal of Malaysia
Author:
Department of Food Science and Technology, Universitas Gadjah Mada, Yogyakarta, Indonesia
Sample:
homogenised colonic tissues
Cited Date:
2024-09-13
Product:
About FineTest® ELISA Kits
What’s the plate size in FineTest® ELISA Kits?

The ELISA plate follows the standard size of microplate: 127.64 mm x 85.60 mm x 14.22 mm(L x W x H).

How about the shelf life and stability of FineTest® ELISA Kits?

Valid for 12 months since the production date. For the shelf life of specific batch number, please check the label printed on the kit. Before delivery, all FineTest® ELISA Kits have been subject to strict quality test.

Which cloned antibodies for FineTest® ELISA Kits are used?

These information is proprietary. Please contact us to learn more about clonality (polyclonality or monoclonality) and host species.

Can I mix reagents from different batches of FineTest® ELISA Kits?

Not suggested. ELISA reagents are optimized for specific batch.

Can FineTest® ELISA Kits be used partially?

Yes. The ELISA plate is dismounted. Enough component volumes are offered by 96T ELISA kit, supporting two groups of standard curve.

How long can the diluted lyophilized standard be stored for continual use?

Used up within 12h.

Can standard curve be extended to any direction?

FineTest® can't support validation of standard concentration outside of standard curve. Ranges of standard curve have been validated among many batches and experimenters, showing stable and accurate performance. The lowest standard concentration is the minimized range for reliable detection results. Adding higher or lower concentration of standard may cause inconsistent signal or false positive.

Sample Issues
Why does detection for serum/plasma sample by FineTest® ELISA Kits require for 1/2 dilution?

Matrix components in serum/plasma can affect detection results. Blocking components in sample dilution buffer can decrease or remove the interference. The dilution can reduce the matrix difference between sample and standard to get better accuracy.

What’s the half-life of protein in serum/plasma/cell culture supernatant?

FineTest® can't determine the half-life of protein in the sample(e.g. serum, plasma or cell culture supernatant). Usually, it's suggested to detect prepared sample immediately or aliquot sample to refrigerate in a disposable container. Avoid freeze-thaw cycle to prevent protein degradation.

What's the expected concentration for particularly analyzing my sample?

Due to the specificity of each sample, it's hard to forecast and depend on sample preparation as well as analytical characteristics. Please contact us to get detection data for reference.

How many samples can FineTest® ELISA Kits detect?

It depends on whether duplicate assay for your sample is required. Such as, you can detect 80 samples without duplication or 40 samples.

Operational Issues
Why is sample loading conducted after equilibrating all kits’ components and samples to room temperature?

Temperature is the important factor for ELISA binding reaction. The reaction of samples at a consistent temperature requires for equilibrating all reagents to room temperature before the assay, including tested samples. Avoid the inaccurate ELISA assay results caused by temperature differences.

Why is duplicate assay suggested for ELISA assay?

To get more accurate assay results, it's strongly suggested to conduct duplicate assay for the standard and sample. Duplicate assay can calculate average value to ensure more accurate assay results and solve drift caused by misoperation during the assay. Calculation of CV evaluates experimental operation and precision of the ELISA kit.

How to get the standard curve with excellent linearity?

Follow the suggested method in the manual to store the standard. Before dissolving the standard, transient centrifugation is required to completely collect the powder. Confirm the fully dissolution and mixing of the standard(about 10min). Then, conduct steps of series dilution. Ensure the fully mixing and accurate pipetting in each step. Properly stop the staining. Choose suitable fitting equation to plot the standard curve.

Can ELISA plates be stacked together for incubation?

To keep the consistent environmental condition of all plates, stacking is not suggested during incubation.

Why is polypropylene tube used for standard dilution in some analyses?

Some proteins or analytes can bind with glass or polystyrene. However, they are not easy to bind with polypropylene tube.

When to stop ELISA reaction?

ELISA assay finally requires for enzyme-catalyzed substrate to complete the staining reaction. Stopping the reaction at the best time is the important factor for successful ELISA assay. When the blue complex appears after adding TMB substrate for 12min, read the O.D. absorbance at 620nm. The staining can be stopped, when the OD value of the darkest color at 620nm is between 0.8 - 0.9. The relevant OD value at 450nm for stopping the staining is between 2.0 - 2.5. If the OD value at 620nm is lower than 0.8, the staining duration can be properly extended.

Why is dual wavelength selected for reading in a microplate reader? What's the purpose of wavelength correction?

Dual wavelength for reading in ELISA assay mainly aims to remove non-specific background interference, and improve accuracy and reliability of the detection. FineTest® usually suggests to set the corrected wavelength as 570nm or 630nm in a microplate reader. The OD value may be higher when directly reading at 450nm. However, the accuracy is lower.

Why is 4 - pl curve fitting is required for generating standard curve?

FineTest® recommends to use ELISA curve fitting software and validate ELISA kit with 4 - pl curve fitting. 4 - pl curve fitting mainly aims to accurately describe the nonlinear dynamic characteristics of antigen - antibody binding. Thus, the quantitative accuracy of low or high concentration of sample is improved. Experimental errors are decreased. The reliability and reproducibility of assay results are ensured.

If sample OD value is higher than OD value of the highest point on the standard curve, what's the suggested solution?

Dilute the sample with dilution buffer and conduct the detection again. Ensure the detected value of the sample falls in the range of standard curve.

Publication Rewards
Are there any rewards to publish papers using FineTest® ELISA Kits?

If you published paper using FineTest® ELISA Kits, you will obtain US$150-US$550 coupon, please contact us for more details.

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