Rat Immunoglobulin G,IgG ELISA Kit

Instructions
Code CSB-E07981r
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

The Rat Immunoglobulin G (IgG) ELISA Kit is specifically designed to standardize the detection of rat IgG in serum and plasma. This kit employs the Competitive-ELISA mechanism to quantitatively measure the concentration of IgG. The rat IgG has been pre-coated on the microtiter plate. Standards and samples are pipetted into the wells with HRP-conjugated antibody specific for rat IgG. A competitive inhibition reaction is launched between IgG (standards or samples) and pre-coated IgG with HRP-conjugated IgG antibody. Following a wash to remove any unbound reagent, the TMB substrate solution is added to the wells. HRP-TMB interaction elicits a chromogenic reaction. The color development is terminated, and the solution color changes from blue to yellow. The color intensity is in opposition to the amount of rat IgG in samples. This kit has been professionally verified with reliable quality, high sensitivity, excellent specificity, good linearity, precision less than 10%, and consistency between batches. See more details on the product instructions.

IgG is the primary type of antibody found in blood circulation extracellular fluids. It has four isoforms, including IgG1, IgG2, IgG3, and IgG4. It is produced and released by plasma B cells in response to an antigen. IgG has four isoforms: IgG1-4. Each IgG has two antigen-binding sites. It protects the body from infections by binding to a variety of pathogens such as fungi, viruses, and bacteria. IgG is the sole Ig subclass that can cross the placenta in humans, and it largely protects the newborn during the first months of life. IgG's relative abundance and excellent specificity to antigens make it become the principal antibody applied in immunological research and clinical diagnostics. The measurement of IgG can be used as a diagnostic tool for certain conditions, such as autoimmune hepatitis if indicated by certain symptoms. And IgG antibodies also play a role in the diagnosis of food allergy.

Target Name Immunoglobulin G,IgG
Alternative Names N/A
Abbreviation IgG
Species Rattus norvegicus (Rat)
Sample Types serum, plasma
Detection Range 31.25 ng/mL-8000 ng/mL
Sensitivity 31.25 ng/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Immunology
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 rat IgG 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)  
1:1000 Average % 100  
Range % 95-104  
1:2000 Average % 90  
Range % 86-95  
1:4000 Average % 87  
Range % 83-92  
1:8000 Average % 101  
Range % 97-106  
Recovery
The recovery of rat IgG 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 88-95  
EDTA plasma (n=4) 93 89-97  
             
             
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    
8000 0.230 0.225 0.228    
2000 0.417 0.407 0.412    
500 0.643 0.664 0.654    
125 1.238 1.257 1.248    
31.25 1.754 1.770 1.762    
0 2.035 1.988 2.012    
Materials provided
  • A 96-well Assay plate --The 96-well plate has been pre-coated with rat IgG.
  • Standard (6 x 0.5 ml)--Dilute the standard at dilution series, read the OD values, and then draw a standard curve.
  • HRP-conjugated IgG(1 x 6 ml) --Bind to the IgG, and HRP catalyzes the TMB to elicit a chromogenic reaction.
  • 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

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