IOCyto Detect™ Rat IL-17F ELISA Kit

CAT#: ITS-0622-CR224
Product Type: Oncology Kit
Target: IL-17F
Short Description
Rat IL-17F ELISA Kit
Homogenous (no wash)
No
Interassay CV
<12%
Intraassay CV
<10%
Detector antibody conjugate
Biotin
Label or dye
HRP
Description
It is designed to detect rat IL-17F in serum, plasma, supernatant. The assay can accurately measure both natural and recombinant rat IL-17F.
Applications
Sandwich ELISA
Target
IL-17F
Reactivity
Rat
Detection Method
Sandwich ELISA
Method Type
ELISA
Sample Type
Serum; plasma and cell culture supernatant
Specificity
Rat
Binding Specificity
Natural and recombinant rat IL-17F
Research Areas
Immuno-oncology
Size
96 Tests
Components
Pre-coated 96 well plate
Standard
Assay Diluent concentrate
Biotinylated Detection Antibody
SAV-HRP
Wash Buffer
Chromogen
Stop Solution
Adhesive Plate Covers
Material not included
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
Sensitivity
0.41 ng/mL
Sample Volume
50 µL
Assay Time
Less than 5 hours
Plate
96 well plate
Reagent Preparation
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.
Assay Procedure
1. For the standard curve, add the standard to the corresponding hole. For sample, add diluted sample to the well.
2. Cover well and incubate at room temperature for 2-4 hours or gently shake at 4°C overnight.
3. Discard the solution and wash it 3-5 times with 1X wash buffer. Use a multi-channel pipette or automatic cleaner to fill each hole with a cleaning buffer. Complete liquid removal at each step is essential for good performance. After the last wash, remove any remaining wash buffer by suction or decanting. Turn the board upside down and drain with a clean paper towel.
4. Add the prepared biotin conjugates into each well.
5. Incubate gently at room temperature for 1-2 hour.
6. Discard the solution. Repeat the washing in Step 3.
7. Add the prepared Streptavidin-HRP solution to each well.
8. Incubate gently at room temperature, shaking for 30-50 minutes.
9. Discard the solution. Repeat the washing in Step 3.
10. Add TMB substrate to each well. The substrate will begin to turn blue.
11. Incubate at room temperature, dark, for 30-45 minutes, shaking gently.
12. Add termination solution to each well. Tap lightly on the sides of the boards to blend. The solution in the hole changes from blue to yellow.
Calculation of Results
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.
Assay Precision
0.41-100 ng/mL
Format
Sandwich ELISA Kit
Precaution of Use
All products are supplied for research and laboratory use only.
Handling Advice
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.
Storage
The ELISA Kits are shipped at 2 to 8°C. Upon receipt, store the kits at 2 to 8°C in dark.
Expiry Date
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.
Note
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.
Shipping
Wet or Dry Ice
Alternative Name
IL17; CTLA8; Cytokine ML-1; IL-17F; Interleukin-17F
Background
Interleukin 17 family (IL17 family) is a family of pro-inflammatory cystine knot cytokines. They are produced by a group of T helper cell known as T helper 17 cell in response to their stimulation with IL-23. Originally, Th17 was identified in 1993 by Rouvier et al. who isolated IL17A transcript from a rodent T-cell hybridoma. The protein encoded by IL17A is a founding member of IL-17 family (see below). IL17A protein exhibits a high homology with a viral IL-17-like protein (O40633) encoded in the genome of T-lymphotropic rhadinovirus Herpesvirus saimiri. In rodents, IL-17A is often referred to as CTLA8.
For Research Use Only | Not For Clinical Use
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