Human lysophosphatidic acid (LPA) Elisa kit

Instructions
Code CSB-EQ028005HU
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

The sensitive quantitative measurement of total human lysophosphatidic acid (LPA) in serum, plasma, and tissue homogenates is easily performed with this 96 well strip format ELISA kit. This assay employs the competitive inhibition enzyme immunoassay technique, in which pre-coated LPA competes with LPA (standards or samples) for the HRP-conjugated antibody specific for LPA. And the color elicited by HRP-TMB chromogenic reaction develops in opposite to the amount of LPA in the sample. The color development is stopped upon the addition of the stop solution and the intensity of the color is measured by a microplate reader at 450 nm.

This Human LPA ELISA Kit has high sensitivity and excellent specificity for detection of LPA. And it also has been validated with precision low than 10%, good linearity, high recovery, and lot-to-lot consistency. Refer to the product instructions for more details.

LPA is a membrane-derived lysophospholipid that can function as a signaling molecule. Upon recognition by GPCRs (LPAR1-LPAR6), LPA mediates in a plethora of biological responses, including differentiation, proliferation, migration, vascular regulation, synaptic plasticity, neurogenesis, cell survival, and cytoskeletal remodeling. Aberrant LPA signaling has been linked to malignant behaviors in various cancers, including breast cancer. Dysregulation of the LPA receptors can lead to hyperproliferation, which may contribute to oncogenesis and metastasis.

Target Name lysophosphatidic acid (LPA)
Abbreviation LPA
Species Homo sapiens (Human)
Sample Types serum, plasma, tissue homogenates
Detection Range 3.9 ng/mL-250 ng/mL
Sensitivity 3.9 ng/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Others
Assay Principle quantitative
Measurement Competitive
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 human LPA 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)  
1:100 Average % 96  
Range % 92-102  
1:200 Average % 94  
Range % 91-98  
1:400 Average % 102  
Range % 98-106  
1:800 Average % 87  
Range % 82-95  
Recovery
The recovery of human LPA 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) 88 80-96  
EDTA plasma (n=4) 96 92-102  
             
             
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    
250 0.090 0.089 0.090    
125 0.147 0.154 0.151    
62.5 0.255 0.267 0.261    
31.25 0.500 0.491 0.496    
15.6 0.757 0.762 0.760    
7.8 1.068 1.059 1.064    
3.9 1.544 1.447 1.496    
0 2.623 2.655 2.639    
Materials provided
  • A 96-well Assay plate --The 96-well plate has been pre-coated with human LPA.
  • Standard(10 x concentrate) (1 x 200 μl) --Dilute the standard at dilution series, read the OD values, and then draw a standard curve.
  • HRP-conjugated LPA antibody(100 x concentrate) (1 x 60 μl) --Bind to the LPAA, and HRP catalyzes the TMB to elicit a chromogenic reaction.
  • HRP-conjugate Diluent (1 x 10 ml) --Dilute the HRP-conjugated LPAantibody solution.
  • Sample Diluent (2 x 20 ml) --Reconstitute the standard and dilute the sample to an appropriate concentration.
  • Wash Buffer (25x concentrate) (1 x 20 ml) --Wash away unbound or free substances.
  • TMB Substrate (1x 10 ml) --Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
  • Stop Solution (1 x 10ml) --Stop the color reaction. The solution color immediately turns from blue to yellow.
  • Four Adhesive Strips (For 96 wells)
  • An Instruction manual
Materials not provided
  • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm - 570 nm.
  • An incubator that can provide stable incubation conditions up to 37°C±5°C.
  • 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

Customer Reviews and Q&A

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 Q&A
Q:

I want to know wihich type of plastic plates are included in this elisa kit to evaluate our optical density.

A:
Thanks for your inquiry!
The plate of our kit CSB-EQ028005HU is with 8 wells× 12 strips.
Pls just use the general microplate reader with the 450nm wavelength.
Any question, pls feel free to contact me.
Q:

CSB-EQ028005HU​ - Human lysophosphatidic acid (LPA) Elisa kit Knowing that LPA is also found in mice, do you think it will also work for mice cells?

A:
Thanks for your inquiry! This kit is specific to human. For different species there is matrix difference, so we don't suggest you use this kit to test mouse samples.

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