Rat Lipopolysaccharides(LPS) ELISA Kit

Instructions
Code CSB-E14247r
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

CUSABIO's rat Lipopolysaccharides(LPS) ELISA kit can assay for rat LPS in the serum, plasma, and tissue homogenates. This assay uses the sandwich ELISA to quantitate LPS in samples. Standards and samples are pipetted into the wells. A highly specific biotin-conjugated LPS antibody is subsequently added to wells. After washing, HRP-avidin conjugate is added to the wells. Following a thorough washing, the TMB substrate solution is added to the wells, making the solution turns blue due to the enzyme reaction catalyzed by HRP. The solution turns blue turns yellow after the addition of the stop solution. The color intensity is measured on a microplate reader at 450nm and is proportional to the amount of LPS captured in the initial step.

This kit provides a detection range of 0.156 ng/mL-10 ng/mL and has high sensitivity as low as 0.039 ng/mL. It has also been validated on several parameters, including precision, linearity, and recovery.

LPS is a large glycolipid containing lipid A, the core oligosaccharide, and the O antigen. It is a major structural component of the outer membrane (OM) of Gram-negative bacteria. LPS transforms the OM into an effective permeability barrier, protecting the membrane from certain kinds of antimicrobial compounds attack. It also plays a critical role in bacteria-host interactions by regulating responses through the immune system.

Target Name Lipopolysaccharides(LPS)
Abbreviation LPS
Species Rattus norvegicus (Rat)
Sample Types serum, plasma, tissue homogenates
Detection Range 0.156 ng/mL-10 ng/mL
Sensitivity 0.039 ng/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Others
Assay Principle quantitative
Measurement Sandwich
Precision
Intra-assay Precision (Precision within an assay): CV%<8%      
Three samples of known concentration were tested twenty times on one plate to assess.  
Inter-assay Precision (Precision between assays): CV%<10%      
Three samples of known concentration were tested in twenty assays to assess.    
             
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of rat LPS in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
  Sample Serum(n=4)  
1:1 Average % 95  
Range % 90-100  
1:2 Average % 102  
Range % 96-107  
1:4 Average % 98  
Range % 90-106  
1:8 Average % 88  
Range % 82-104  
Recovery
The recovery of rat LPS spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.
Sample Type Average % Recovery Range  
Serum (n=5) 93 84-100  
EDTA plasma (n=4) 104 100-108  
             
             
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
ng/ml OD1 OD2 Average Corrected  
10 1.987 1.999 1.993 1.819  
5 1.395 1.357 1.376 1.202  
2.5 0.887 0.897 0.892 0.718  
1.25 0.595 0.576 0.586 0.412  
0.625 0.391 0.384 0.388 0.214  
0.312 0.301 0.324 0.313 0.139  
0.156 0.247 0.261 0.254 0.080  
0 0.173 0.175 0.174    
Materials provided
  • A micro ELISA plate ---The 96-well plate has been pre-coated with an anti-Rat LPS antibody.
  • Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
  • One vial Biotin-labeled LPS antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
  • One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
  • One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody.
  • One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
  • One vial Sample Diluent (50 ml/bottle) ---Dilute the sample to an appropriate concentration.
  • One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
  • One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
  • One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
  • Four Adhesive Strips (For 96 wells) ---Cover the microplate when incubating.
  • An instruction manual
Materials not provided
  • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
  • An incubator can provide stable incubation conditions up to 37°C±5°
  • Centrifuge
  • Vortex
  • Squirt bottle, manifold dispenser, or automated microplate washer
  • Absorbent paper for blotting the microtiter plate
  • 50-300ul multi-channel micropipette
  • Pipette tips
  • Single-channel micropipette with different ranges
  • 100ml and 500ml graduated cylinders
  • Deionized or distilled water
  • Timer
  • Test tubes for dilution
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 3-5 working days

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 Q&A
Q:

I am interested in your kit Rat Lipopolysaccharides, LPS ELISA Kit (CSB-E14247r) (24 strip-well).
Taking into consideration the number of standards, how many samples can you analyse? Do you need to do this in duplicate?

A:
Thanks for your inquiry.
CSB-E14247r
For this target, dilution is not needed if you test serum and plasma samples. We validated normal serum before and the test value is ND--0.25ng/ml. If you test samples of other modes, you can refer to the research situation of this mode and judge if pretest is needed or not. Take 24T as an example, if you run single well for the standard and the sample, you can test 16 samples. You can also run duplicated wells to reduce errors during the assay.
Pls let me know if you have any further questions. Thank you.

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