IOCyto Detect™ IL-12 p40 Rat ELISA Kit (CAT#: ITS-0622-CR238)

96 Tests

Pre-coated 96 well plate
Standard
Assay Diluent concentrate
Biotinylated Detection Antibody
SAV-HRP
Wash Buffer
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

<3 pg/mL

10 µL to 100 µL

Less than 4 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. 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.

1. Read the absorbance at 450 nm. Read the plate within 2 hours after adding the Stop Solution. 2. Use curveȬfitting software to generate the standard curve. A 4 parameter algorithm provides the best standard curve fit. Optimally, the background absorbance may be subtracted from all data points, including standards, unknowns and controls, prior to plotting. 3. Read the concentrations for unknown samples and controls from the standard curve. Multiply value(s) obtained for sample(s) by the appropriate factor to correct for the sample dilution.

15.6-1000 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 Ice

IL-12 subunit p40; IL-12B; IL12; subunit p40; interleukin 12; p40; interleukin 12B (natural killer cell stimulatory factor 2; cytotoxic lymphocyte maturation factor 2; p40); interleukin-12 beta chain; Interleukin-12 subunit beta; natural killer cell stimulatory factor; 40 kD subunit; NK cell stimulatory factor chain 2; NKSF2

Interleukin 12 (IL-12) is an interleukin that is naturally produced by dendritic cells, macrophages, neutrophils, and human B-lymphoblastoid cells (NC-37) in response to antigenic stimulation. IL-12 belongs to the family of interleukin-12. IL-12 family is unique in comprising the only heterodimeric cytokines, which includes IL-12, IL-23, IL-27 and IL-35. Despite sharing many structural features and molecular partners, they mediate surprisingly diverse functional effects.