Human BAFF/CD257(B-Cell Activating Factor) ELISA Kit

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  • sandwich-Ab-ELISA-Elabscience
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    • sandwich-Ab-ELISA-Elabscience
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    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
      Catalog number:E-EL-H0009

      Synonyms:TNFSF13B, BLYS, TALL1, THANK, TNFSF20, ZTNF4

      Size:
      • 96T
      • 96T*5
      Qty:
      - +
      Price: $580

      Reactivity: Human

      Detection Range: 62.50~4000 pg/mL

      Sensitivity: 37.50 pg/mL

      Lead Time: 7~10 days

      Add to cart Compare Bulk request Manual

      Order directly online from USA, please contact us firstly if order from the other countries.

      Intended use

      This ELISA kit can be applied to the in vitro quantitative determination of Human BAFF/CD257 concentrations in serum, plasma and other biological fluids.

      Test principle

      This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human BAFF/CD257. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human BAFF/CD257 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human BAFF/CD257, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human BAFF/CD257. The concentration of Human BAFF/CD257 in samples can be calculated by comparing the OD of the samples to the standard curve.

      Assay type Sandwich-ELISA
      Format 96T
      Assay time 4.5h
      Reactivity Human
      Detection Method Colormetric
      Detection Range 62.50—4000 pg/mL
      Sensitivity 37.50 pg/mL
      Sample Volume Required Per Well 100 μL
      Sample Type Serum, plasma and other biological fluids

      Specificity

      This kit recognizes some recombinant and natural Human BAFF/CD257. No significant cross-reactivity or interference was observed.

      Typical data

      For convenience in result calculation, absorbance can be used as ordinate and standard concentrations as abscissa. The standard curve provided in the manual is only for reference, and experimenters should draw the standard curve based on data of themselves.

      (pg/mL) O.D Average Corrected
      4000 2.258
      2.28
      2.269 2.192
      2000 1.517
      1.541
      1.529 1.452
      1000 0.889
      0.887
      0.888 0.811
      500 0.466
      0.474
      0.47 0.393
      250 0.274
      0.254
      0.264 0.187
      125 0.187
      0.159
      0.173 0.096
      62.50 0.119
      0.135
      0.127 0.05
      0 0.072
      0.082
      0.077 --

      Precision

      Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high leve Human BAFF/CD257 were tested 20 times on one plate, respectively.
      Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Human BAFF/CD257 were tested on 3 different plates, 20 replicates in each plate.

      Intra-assay Precision Inter-assay Precision
      Sample 1 2 3 1 2 3
      n 20 20 20 20 20 20
      Mean
      (pg/mL)
      185.79 600.16 1343.28 173.82 543.69 1425.24
      Standard
      deviation
      11.65 35.47 54.13 11.94 28.71 45.04
      C V (%) 6.27 5.91 4.03 6.87 5.28 3.16

      Recovery

      The recovery of Human BAFF/CD257 spiked to three different levels in samples throughout the range of the assay in various matrices was evaluated.

      Sample Type Range (%) Average Recovery (%)
      Serum (n=5) 87-101 94
      EDTA plasma (n=5) 86-100 91
      Cell culture media (n=5) 90-105 96

      Linearity

      Samples were spiked with high concentrations of Human Abbre and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

      Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
      1:2 Range (%) 92-105 95-108 91-103
      Average (%) 100 101 97
      1:4 Range (%) 91-108 86-99 87-102
      Average (%) 99 93 94
      1:8 Range (%) 86-99 85-97 85-100
      Average (%) 93 90 91
      1:16 Range (%) 91-106 86-100 85-96
      Average (%) 98 92 90

      Kit components & Storage

      The unopened kit can be stored at 4℃ for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions since the kit is received.

      Item Specifications Storage
      Micro ELISA Plate(Dismountable) 8 wells ×12 strips -20℃, 6 months
      Reference Standard 2 vials
      Concentrated Biotinylated Detection Ab (100×) 1 vial, 120 μL
      Concentrated HRP Conjugate (100×) 1 vial, 120 μL -20℃(shading light), 6 months
      Reference Standard & Sample Diluent 1 vial, 20 mL 4℃, 6 months
      Biotinylated Detection Ab Diluent 1 vial, 14 mL
      HRP Conjugate Diluent 1 vial, 14 mL
      Concentrated Wash Buffer (25×) 1 vial, 30 mL
      Substrate Reagent 1 vial, 10 mL 4℃(shading light)
      Stop Solution 1 vial, 10 mL 4℃
      Plate Sealer 5 pieces
      Product Description 1 copy
      Certificate of Analysis 1 copy

      Note: All reagent bottle caps must be tightened to prevent evaporation and microbial pollution.
      The volume of reagents in partial shipments is a little more than the volume marked on the label, please use in measuring instead of directly pouring.

      Other supplies required

      • Microplate reader with 450nm wavelength filter
      • High-precision transferpettor, EP tubes and disposable pipette tips
      • 37℃ Incubator
      • Deionized or distilled water
      • Absorbent paper
      • Loading slot for Wash Buffer

      Assay Procedure

      1.Add 100 μL of standard or sample to each well.

      2.Remove the liquid.

      3.Add 100 μL Biotinylated Detection Ab. Incubate 1 hour at 37℃.Aspirate and wash 3 times.

      4.Add 100 μL HRP Conjugate. Incubate 30 min at 37℃.Aspirate and wash 5 times.

      5.Add 90 μL of Substrate Reagent. Incubate for 15 min at 37℃.

      6.Add 50 μL of Stop Solution.

      7.Read the OD at 450 nm immediately. Calculation of the results.

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