IOCyto Detect™ TNF alpha Equine Uncoated ELISA Kit (CAT#: ITS-0622-CR196)

5 x 96 Tests

Capture Antibody
Detection Antibody
Standard
ELISA/ELISPOT Coating Buffer
Assay Diluent
Detection enzyme
Substrate Solution
Certificate of Analysis
96 Well Plate

96-well ELISA plate reader (microplate spectrophotometer)
ELISA plate washer
(Optional) Microplate shaker
Pipettes and pipettors
Refrigerator
96-well plate

<15.6 pg/mL

100 µL

Less than 36 hours

96 well plate

1. Dilute the Coating Antibody 1:100 in carbonate-bicarbonate buffer.
2. Add diluted Coating Antibody to each well.
3. Aspirate Coating Antibody solution and add Blocking Buffer to each well.
4. Aspirate Blocking Buffer and proceed to assay or allow to dry overnight at room temperature.

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.

Calculate results using graph paper or curve-fitting statistical software. The horse TNFα amount in each sample is determined by interpolating from the absorbance value (Y axis) to horse TNFα concentration (X axis) using the standard curve.

15.6-1000 pg/mL

Uncoated 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.

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

TNF; DIF; TNF-alpha; TNFA; TNFSF2; Tumour necrosis factor; TNF-α; tumor necrosis factor; TNLG1F; Tumor necrosis factor alpha

Tumor necrosis factor (TNF, cachexin, or cachectin; often called tumor necrosis factor alpha or TNF-α) is an adipokine and a cytokine. TNF is a member of the TNF superfamily, which consists of various transmembrane proteins with a homologous TNF domain. As an adipokine, TNF promotes insulin resistance, and is associated with obesity-induced type 2 diabetes. As a cytokine, TNF is used by the immune system for cell signaling. If macrophages (certain white blood cells) detect an infection, they release TNF to alert other immune system cells as part of an inflammatory response.