Human I-TAC ELISA development kit contains the key components required for the quantitative measurement of natural and/or recombinant I-TAC in a sandwich ELISA format within the range of 32-2,000 pg/mL. Using the ELISA protocol described below, this kit provides sufficient reagents to assay I-TAC in approximately 1,500 ELISA plate wells.
Contents of Kit
1. PBS: Dilute 10xPBS to 1xPBS, pH 7.2 in sterile water.
2. Wash Buffer: 0.05% Tween-20 in PBS
3. Block Buffer: 1.0% BSA in PBS
4. Diluent: 1.0% BSA in PBS
5. ELISA microplates (Thermo Fisher Cat. # 456529);
6. BSA (Sigma Cat. # A-7030);
7. Stop Solution 2 M Sulfuric Acid (Sigma Cat. # 339741);
8. Dulbecco's PBS [10x] (Gibco BRL Cat. # 14200-075).
Unopened Kit Store at 2-8°C. Do not use past kit expiration date. For more detailed information, please download the document on our website.
Chemokine (C-X-C motif) ligand 11 (CXCL11) is a small cytokine belonging to the CXC chemokine family that is also called Interferon-inducible T-cell alpha chemoattractant (CXCL11) and Interferon-gamma-inducible protein 9 (IP-9). It is highly expressed in peripheral blood leukocytes, pancreas and liver, with moderate levels in thymus, spleen and lung and low expression levels were in small intestine, placenta and prostate.
Reconstitution And Storage
Capture Antibody: 180 μg of antigen-affinity purified goat anti-I-TAC. Centrifuge vial prior to opening. Reconstitute in 1.0 mL PBS for a concentration of 180 μg/mL. Following reconstitution the Capture antibodies may be stored at 2-8°C for up to 6 months. For long term storage it is recommended to aliquot into working volumes and store at -70°C in a manual defrost freezer.
Detection Antibody: 36 μg of biotinylated antigen-affinity purified goat anti-I-TAC. Centrifuge vial prior to opening. Reconstitute in 1.0 mL sterile 1.0% BSA in PBS for a concentration of 36 μg/mL. Following reconstitution the Detection antibodies may be stored at 2-8°C for up to 6 months. For long term storage it is recommended to aliquot into working volumes and store at -70°C in a manual defrost freezer.
Human I-TAC Standard: 27.5 ng of recombinant I-TAC. Centrifuge vial prior to opening. Reconstitute in 0.5 mL sterile 1.0% BSA in PBS for a concentration of 55 ng/mL. The Standard may be stored at 2-8°C for one month or aliquoted and stored at -70°C for up to three months in a manual defrost freezer.
UltraAvidin-HRP Conjugate: 40 μL vial. Upon receipt, UltraAvidin-HRP conjugate should be stored at 2-8°C, DO NOT FREEZE.
TMB Liquid Substrate: Aspirate and wash plate 4 times. Add 100 μL of TMB HRPO Microwell Substrate Standard Kinetic One Component "Ready Use" to each well. Incubate at room temperature and monitor color development with an ELISA plate reader at 450 nm with wavelength correction set at 540 nm or 570 nm. The reaction may be stopped after 15-20 minutes by adding 100 μL of 2 M sulfuric acid to each well.