Phone:
+1 877 302 8632
Fax:
+1 888 205 9894 (Toll-free)
E-Mail:
orders@antibodies-online.com

CD28 ELISA Kit (CD28) ELISA Kit

CD28 Reactivity: Human Colorimetric Sandwich ELISA 0.63 ng/mL - 40 ng/mL Cell Culture Supernatant, Plasma, Serum
Catalog No. ABIN6962349
$638.00
Plus shipping costs $45.00
96 tests
local_shipping Shipping to: United States
Delivery in 8 to 11 Business Days
  • Target
    CD28
    Reactivity
    • 7
    • 3
    • 3
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Human
    Detection Method
    Colorimetric
    Method Type
    Sandwich ELISA
    Detection Range
    0.63 ng/mL - 40 ng/mL
    Minimum Detection Limit
    0.63 ng/mL
    Application
    ELISA
    Purpose
    The kit is a sandwich enzyme immunoassay technique for the in vitro quantitative measurement in various sample types.
    Sample Type
    Cell Culture Supernatant, Plasma, Serum
    Analytical Method
    Quantitative
    Specificity
    This kit recognizes Human CD28 in samples. No Significant cross-reactivity or interference between Human CD28 and analogues was observed.
    Sensitivity
    0.38 ng/mL
    Components
    • Pre-coated, ready to use 96-well strip plate, flat buttom
    • Plate sealer for 96 wells
    • Reference Standard
    • Reference Standard & Sample Diluent
    • Biotinylated Detection Antibody (100 x concentrate)
    • HRP Conjugate (100 x concentrate)
    • Biotinylated Detection Antibody Diluent
    • HRP Conjugate Diluent
    • Substrate Reagent
    • Stop Solution
    • Wash Buffer (25 x concentrate)
    • Instruction manual
  • Sample Volume
    100 μL
    Assay Time
    3.5 h
    Plate
    Pre-coated
    Protocol
    1. Add 100 µL standard or sample to each well. Incubate for 90 min at 37 °C.
    2. Remove the liquid. Add 100 µL Biotinylated Detection Antibody. Incubate for 1 hour at 37 °C.
    3. Aspirate and wash 3 times.
    4. Add 100 µL HRP Conjugate. Incubate for 30 min at 37 °C.
    5. Aspirate and wash 5 times.
    6. Add 90 µL Substrate Reagent. Incubate for 15 min at 37 °C.
    7. Add 50 µL Stop Solution. Read at 450 nm immediately.
    8. Calculation of results.
    Reagent Preparation
    1. Bring all reagents to room temperature (18~25 °C) before use. Follow the Microplate reader manual for set-up and preheat it for 15 min before OD measurement.
    2. Wash Buffer: Dilute 30 mL of Concentrated Wash Buffer with 720 mL of deionized or distilled water to prepare 750 mL of Wash Buffer.Note: if crystals have formed in the concentrate, warm it in a 40 °C water bath and mix it gently until the crystals have completely dissolved
    3. Standard working solution: Centrifuge the standard at 10,000xg for 1 min. Add 1.0 mL of Reference Standard &Sample Diluent, let it stand for 10 min and invert it gently several times. After it dissolves fully, mix it thoroughly with a pipette. This reconstitution produces a working solution of 40 ng/mL. Then make serial dilutions as needed. The recommended dilution gradient is as follows: 40, 20, 10, 5, 2.5, 1.25, 0.63, 0 ng/mL. Dilution method: Take 7 EP tubes, add 500 μLof Reference Standard & Sample Diluent to each tube. Pipette 500 μLof the 40 ng/mL working solution to the first tube and mix up to produce a 20 ng/mL working solution. Pipette 500 μLof the solution from the former tube into the latter one according to these steps. The illustration below is for reference. Note: the last tube is regarded as a blank. Don't pipette solution into it from the former tube.
    4. Biotinylated Detection Antibody working solution: Calculate the required amount before the experiment (100 μL/well). In preparation, slightly more than calculated should be prepared. Centrifuge the stock tube before use, dilute the 100x Concentrated Biotinylated Detection Antibody to 1xworking solution with Biotinylated Detection Antibody Diluent.
    5. Concentrated HRP Conjugate working solution: Calculate the required amount before the experiment (100 μL/well). In preparation, slightly more than calculated should be prepared. Dilute the 100x Concentrated HRP Conjugate to 1x working solution with Concentrated HRP Conjugate Diluent.
    Assay Precision
    Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human CD28 were tested 20 times on one plate, respectively.
    Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human CD28 were tested on 3 different plates, 20 replicates in each plate.
    Both intra-CV and inter-CV are < 10 %.
    Restrictions
    For Research Use only
  • Storage
    4 °C,-20 °C
    Expiry Date
    6 months
  • Target
    CD28
    Alternative Name
    Cluster of Differentiation 28 (CD28 ELISA Kit Abstract)
    Synonyms
    Tp44, CD28RNA, cd28, CD28 molecule, CD28 antigen, Cd28 molecule, CD28 molecule L homeolog, CD28, Cd28, cd28.L
    Pathways
    TCR Signaling, Fc-epsilon Receptor Signaling Pathway, EGFR Signaling Pathway, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response
You are here:
help Support