IOCyto Detect™ Rabbit IL-8 ELISA Kit (CAT#: ITS-0622-CR97)

96 Tests

Pre-coated 96 well plate
Standard
Sample Diluent
Assay Buffer concentrate
Biotinylated Detection Antibody
SAV-HRP
Wash Buffer
Controls
Chromogen
Stop Solution
Adhesive Plate Covers

Distilled or deionized water
Microtiter Plate Reader software capable of measuring at 450 nm
Dishwasher - Automatic or Manual (Dispenser)
Calibrate adjustable precision pipettes and glass or plastic tubes to dilute solutions

0.8 pg/mL

50 µL

Less than 5 hours

96 well plate

1. Prepare Wash Buffer
Allow wash buffer concentrate to reach room temperature and mix to redissolve any precipitated salts. Dilute Wash Buffer Concentrate into deionized or distilled water. Store the concentrate and 1X wash buffer in the refrigerator.
2. Prepare the thinner
The test diluent should be diluted with deionized or distilled water before use.
3. Preparation of Biotin Conjugation
Spin the biotin conjugate briefly before use. Gently stir up and down with a pipette. Collect samples in pyrogen/endotoxin-free tubes. If samples cannot be tested immediately after collection, freeze samples. Freeze samples to avoid multiple freeze-thaw cycles.
4. Pre-dilute samples
For different types of samples, different dilution solutions should be used for dilution.
5. Prepare 1X Streptavidin-HRP Solution
Quickly swirl Streptavidin-HRP before use and mix gently up and down with a pipette as a precipitate may form during storage. Do not store the diluted solution for future use.
6. Dilution Standards
Dilute standards using glass or plastic tubes. Simply spin off a vial of lyophilized standards.

1. Add Incubation Buffer to all wells except the chromogen blank.
2. Add the standard, buffer solution, or cell culture medium sample to the appropriate wells. for
Serum and control samples, add standard dilution buffer, then add samples to
suitable well. Leave the chromogen blank holes blank.
3. Tap the side of the plate to mix. Cover the plate with a plate lid and incubate at room temperature for 2-3 hours.
4. Aspirate the solution thoroughly and wash the wells 4 times with wash buffer.
5. Add Biotin Conjugate solution to each well except the chromogen blank.
6. Cover the plate and incubate at room temperature for 1-2 hours.
7. Aspirate the solution thoroughly and wash the wells 4 times with wash buffer.
8. Add Streptavidin-HRP solution to each well except the chromogen blank.
9. Cover the plate with a plate cover and incubate at room temperature.
10. Aspirate the wells thoroughly and wash the wells 4-6 times with wash buffer.
11. Add stable chromogen to each well. The substrate solution begins to turn blue.
12. Incubate at room temperature for 30-45 minutes in the dark.
13. Add stop solution to each well. Tap the side of the plate to mix. The solution in the well changed from blue to yellow.

1. Read the absorbance at 450nm. Read the plate within 30 minutes of adding the stop solution.
2. Generate a standard curve using curve fitting software.
3. Read the concentrations of unknown samples and controls from the standard curve. The sample dilution is corrected by an appropriate factor, resulting in a number of values (s) for the sample.

0.8-200 pg/mL

Sandwich ELISA Kit

All products are supplied for research and laboratory use only.

All blood components and biological materials should be treated with precautions as potentially hazardous. Strictly follow the management principles of the Centers for Disease Control and Prevention, Occupational Safety and Health Administration for handling and disposing of infectious agents.

The ELISA Kits are shipped at 2 to 8°C. Upon receipt, store the kits at 2 to 8°C in dark.

Stability If properly stored, all components are stable for up to 12 months. For expiry dates for the entire kit, see the kit label. The expiration date of each ingredient is shown on the bottle label. The expiration date of a kit ingredient is guaranteed only if the ingredient is properly stored and, in the event of repeated use of an component, the reagent will not be contaminated by the first treatment.

Each production lot of this ELISA kit is quality tested to meet criteria such as sensitivity, specificity, precision and lot-to-lot consistency. See the manual for more information on validation.

Wet or Dry Ice

CXCL8; chemokine (C-X-C motif) ligand 8; GCP-1; GCP1; LECT; LUCT; LYNAP; MDNCF; MONAP; NAF; NAP-1; NAP1; IL8; C-X-C motif chemokine ligand 8; Interleukin-8; SCYB8

Interleukin 8 (IL-8 or chemokine (C-X-C motif) ligand 8, CXCL8) is a chemokine produced by macrophages and other cell types such as epithelial cells, airway smooth muscle cells and endothelial cells. Endothelial cells store IL-8 in their storage vesicles, the Weibel-Palade bodies. In humans, the interleukin-8 protein is encoded by the CXCL8 gene. IL-8 is initially produced as a precursor peptide of 99 amino acids which then undergoes cleavage to create several active IL-8 isoforms. In culture, a 72 amino acid peptide is the major form secreted by macrophages.