Human anti-double stranded DNA(dsDNA) antibody(IgG) ELISA Kit

Instructions
Code CSB-E04911h
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

The product CSB-E04911h is a ready-to-use microwell, strip plate ELISA Kit for quantitative analysis of human anti-double-stranded DNA (dsDNA) antibody (IgG) (dsDNA Ab (IgG)) in serum. Based on the Indirect-ELISA technique in combination with enzyme-substrate chromogenic reaction, the dsDNA Ab (IgG) levels can be calculated referring to the standard curve. This Human dsDNA Ab (IgG) ELISA Kit has been verified by multiple tests with high sensitivity, strong specificity, precision less than 15%, good linearity, and lot-to-lot consistency.

This assay employs the qualitative enzyme immunoassay technique. Samples are pipetted into the wells with anti-human IgG conjugated HRP. Any antibodies specific for the antigen present will bind to the pre-coated antigen-specific for dsDNA Ab (IgG) on the microwell plate. Following a wash to remove any unbound reagent, substrate A and B solutions are added to the wells and color develops in proportion to the amount of human dsDNA Ab (IgG) bound in the initial step. The color development is terminated and the intensity of the color is measured.

Anti-dsDNA antibody (IgG) is an isoform of anti-dsDNA antibodies that are related to active systemic lupus erythematosus (SLE) and lupus nephritis (LN). Anti-dsDNA antibodies recognize diverse DNA structures, including single-stranded (ss)DNA, Z-DNA, bent or elongated dsDNA, DNA: RNA hybrids, locked-nucleic acids, and peptide: nucleic acid hybrids. Xiaoyu Wang and Yumin Xia demonstrated that anti-dsDNA IgG contributes to renal fibrosis by downregulating suppressor of cytokine signaling 1 (SOCS1) and activating Jak-STAT1 signals, which modulate the expression of profibrotic genes, TGF-β1, platelet-derived growth factor B, and connective tissue growth factor.

Target Name anti-double stranded DNA(dsDNA) antibody(IgG)
Alternative Names N/A
Abbreviation dsDNA Ab (IgG)
Species Homo sapiens (Human)
Sample Types serum
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Autoimmunity
Assay Principle qualitative
Measurement Indirect
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.    
             
Typical Data
             
             
Test parameter specification test result  
Positive control >0.8 1.246  
Negative control <0.2 0.053  
Positive rate 10,Positive 100%  
Negative rate 10,Negative 100%  
                   
Materials provided
  • A micro ELISA plate --- The 96-well plate has been pre-coated with antigen-specific for dsDNA Ab (IgG). This dismountable microplate can be divided into 12 x 8 strip plates.
  • Negative Control (1 x 800 μl)
  • Positive Control (1 x 800 μl)
  • HRP-conjugate (1 x10 ml) --- Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
  • Sample Diluent (1 x 20 ml) --- Dilute the standard and sample solution.
  • Wash Buffer (25 x concentrate) (1 x20 ml) --- Wash away unbound or free substances.
  • Substrate A 1 x 5 ml --- Mix with substrate B, and the TMB is catalyzed by HRP to elicit a chromogenic signal.
  • Substrate B 1 x 5 ml --- Mix with substrate A, and the TMB is catalyzed by HRP to elicit a chromogenic signal.
  • Stop Solution (1 x 5 ml) --- Stop the color reaction. The solution color immediately turns from blue to yellow.
  • Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
  • 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 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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