LPS(Lipopolysaccharides) ELISA Kit

This kit is based on Competitive-ELISA detection method and takes 2h assay time. The microplate provided in this kit has been precoated with anti LPS antibody. Sample or standard completely mixes with fixed amount of biotin-labeled LPS working solution and also add into each well. LPS in the sample or standard competes with biotin-labeled LPS to bind with antibody precoated on the microplate. Free components are washed off. After adding HRP-Streptavidin Conjugate (SABC), wash off unbound conjugate. 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. Compare OD450 value of sample with the standard curve through curve fitting software to determine the concentration of LPS in the sample. The concentration of the target substance is inversely proportional to the OD450 value.

Catalogue No.: 
EU3126
Alias: 
LPS ELISA Kit, Lipopolysaccharides ELISA Kit
Species: 
Universal
Range: 
0.313-20ug/ml
Sensitivity: 
0.188ug/ml
  • SPECIFICATIONS
  • CITATIONS
Product Name
LPS(Lipopolysaccharides) ELISA Kit
Alias
LPS ELISA Kit, Lipopolysaccharides ELISA Kit
Catalogue No.
EU3126
Size
48T/96T
Species
Universal
Sample Type
Serum, Plasma, Cell Culture Supernatant, cell or tissue lysate, Other liquid samples
Detection Method
Competitive ELISA, Coated with Antibody
Detection Wavelength
OD450
Reaction Duration
2 hours
Range
0.313-20ug/ml
Sensitivity
0.188ug/ml
Storage
2-8°C(Sealed), Don't cryopreserve.
Specificity
Specifically binds with LPS , 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 Lyophilized Biotin-labeled Antigen(Concentrated) 1vial 1vial
E034 HRP-Streptavidin Conjugate(SABC, 100X) 60ul 120ul 2-8°C (Avoid Direct Light)
E024 TMB Substrate 5ml 10ml
E005 10mM PBS 200ul 200ul 2-8°C
E039 Sample Dilution Buffer 10ml 20ml
E040 Antigen 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: Wash the plate twice before adding the standard and sample.
  • Step 2: Add 100ul standard/sample/biotin-labeled antigen working solution into each well. Seal the plate and statically incubate for 45 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 into each well, seal the plate and statically incubate for 30 minutes at 37°C.
  • Washing:
  • 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.(ug/ml) OD-1 OD-2 Average
0 2.334 2.453 2.381
0.312 1.734 1.822 1.769
0.625 1.437 1.51 1.466
1.25 1.266 1.33 1.291
2.5 1.072 1.126 1.094
5 0.826 0.868 0.843
10 0.715 0.752 0.73
20 0.676 0.71 0.689
EU3126 Standard Curve Image
Recovery

Add a certain amount of LPS into the sample. Calculate the recovery by comparing the measured value with the expected amount of LPS in the sample.

Sample Type Recovery Range(%) Average(%)
serum(n=10) 88-103 96
EDTA plasma(n=10) 90-102 95
Heparin plasma(n=10) 92-104 94
Linearity

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

Sample Type 1:2 1:4 1:8
serum(n=10) 85-103% 96-101% 82-93%
EDTA plasma(n=10) 83-99% 82-100% 83-92%
Heparin plasma(n=10) 85-96% 84-100% 93-105%
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 (ug/ml) 0.62 2.6 10.38 0.62 2.51 10.06
Standard deviation 0.03 0.13 0.34 0.03 0.12 0.63
CV(%) 4.68 4.88 3.28 5.23 4.96 6.24
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: 7.49
Journal:
Fertility and Sterility
Author:
Department of Obstetrics and Gynecology, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, People’s Republic of China
Cited Date:
2023-05-25
Product:
IF: 4.9
Journal:
Probiotics and Antimicrobial Proteins
Author:
Graduate Program in Cardiovascular Sciences, Fluminense Federal University (UFF), Niterói, RJ, Brazil
Cited Date:
2023-11-10
Product:
IF: 3.5
Journal:
Journal of the American Nutrition Association
Author:
Cardiovascular Sciences Post Graduate Program, Fluminense Federal University (UFF), Niter贸i, Rio de Janeiro, Brazil
Cited Date:
2023-09-01
Product:
IF: 2.22
Journal:
Animal Reproduction Science
Author:
Ankara University, Faculty of Veterinary Medicine, Department of Obstetrics and Gynecology, 06110 Ankara, Turkiye
Cited Date:
2023-06-16
Product: