Fish immunoglobulin M (IgM) ELISA Kit

Instructions
Code CSB-E12045Fh
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

The fish immunoglobulin M (IgM) ELISA kit is suitable for the quantitative measurement of the IgM levels in fish serum, plasma, or tissue homogenates. It provides a detection range of 1.25 μg/mL-50 μg/mL. It also has high sensitivity, excellent specificity, high precision, good linearity, and high recovery.

This assay employs the competitive inhibition enzyme immunoassay technique using an anti-fish IgM antibody and an HRP-conjugated IgM. In this assay, the microplate has been pre-coated with a goat-anti-rabbit antibody. Samples or standards are pipetted into wells with the anti-fish IgM antibody and HRP-conjugated IgM. IgM in the samples or the standards and HRP-conjugated IgM competes with binding to the anti-fish IgM antibody. The solution appears blue after the addition of the TMB substrate. The color reaction is terminated after adding the stop solution. The color intensity is inverse to the amount of IgM in the sample.

IgM is the initial antibody produced by B cells in response to a foreign pathogen. It is rapidly generated during a primary immune response. It usually lasts for about 3 weeks, peaks in the second week, and declines gradually till it disappears. IgG emerges at this time. IgM not only acts as the first line of host defense against infections but also plays an important role in immune modulation and immunological tolerance.

Target Name immunoglobulin M
Abbreviation IgM
Species Fish
Sample Types serum, plasma, tissue homogenates
Detection Range 1.25 μg/mL-50 μg/mL
Sensitivity 1.25 μg/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%<15%      
Three samples of known concentration were tested twenty times on one plate to assess.  
Inter-assay Precision (Precision between assays): CV%<15%      
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 fish IgM 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 % 93  
Range % 86-101  
1:2 Average % 97  
Range % 90-104  
1:4 Average % 95  
Range % 84-105  
1:8 Average % 95  
Range % 91-99  
Recovery
The recovery of fish IgM 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) 97 92-105  
EDTA plasma (n=4) 95 91-102  
             
             
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
μg/ml OD1 OD2 Average    
50 0.122 0.124 0.123    
25 0.213 0.199 0.206    
11.5 0.401 0.427 0.414    
3.75 0.745 0.761 0.753    
1.25 1.375 1.402 1.389    
0 2.541 2.532 2.537    
Materials provided
  • A 96-well Assay plate --The 96-well plate has been pre-coated with goat-anti-rabbit antibody.
  • Standard (6 x 0.5 ml) --Dilute the standard at dilution series, read the OD values, and then draw a standard curve.
  • HRP-conjugated IgM (1 x 6 ml) --Bind to the IgM antibody, and HRP catalyzes the TMB to elicit a chromogenic reaction.
  • IgM Antibody (1 x 6 ml) --Interact with the IgM.
  • 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 600 nm - 630 nm.
  • An incubator that 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

Citations

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 Q&A
Q:

CUSABIO fish immnoglobulin M (IgM) ELISA kit Does this kit can be used for separate experiments?around 30-40 wells every time?

A:
Thanks for your inquiry.
#CSB-E12045Fh,Fish immunoglobulin M (IgM) ELISA Kit
YES,it can.The plate is Breakapart and can be used for seperate experiments.You can put the break part in the
Aluminum foil bag for next use.The manual is provided to you by e-mail.Additionally,we received 21 published papers until now.You can refer to it. Pls let me know if you have any further questions. Thank you.
Q:

Regarding your ELISA kit CSB-E12045Fh, could you please advise on the following:
1. Capture Antibody: host animal species, clonality
2. Detection Antibody: host animal species, clonality
3. Standard
Also, what sample types have been tested with this kit, and what were the reference values?

A:
Thanks for your inquiry.
CSB-E12045Fh
The target is a special protein and there is no uniprot.
This kit is based on competitive method. The plate is coated with goat-anti-rabbit antibody. The specific antibody is rabbit polyclonal antibody. The standard is purified crude extract of natural samples.
We tested undiluted serum and plasma before and the test value is 1.25--3.75μg/ml.
Pls let me know if you have any further questions. Thank you.

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