The Rat TNF-alpha ELISA Kit (SKU: GXEL00510) is a quantitative sandwich ELISA for the measurement of Rat TNF-alpha in cell culture supernates, serum, and plasma. The assay has a detection range of 15.6 - 1,000 pg/mL and a sensitivity of 5.21 pg/mL.
This kit is based on a quantitative sandwich enzyme-linked immunosorbent assay (ELISA). Standards and samples are added to the pre-coated microplate wells, where Rat TNF-alpha participates in a format-specific immunoreaction. After the required incubation and washing steps, enzyme-conjugated reagent and substrate solution are added sequentially to generate a measurable colorimetric signal. The optical density (OD) value is positively correlated with the concentration of Rat TNF-alpha in the sample. The concentration of Rat TNF-alpha is calculated by interpolation from the standard curve.
Product Name:
Rat TNF-alpha ELISA Kit
SKU:
GXEL00510
Size:
96 Assays
Reactivity:
Rat
Assay Type:
Sandwich
Detection Method:
Colorimetric (450 nm)
Sensitivity:
5.21 pg/mL
Range:
15.6 - 1,000 pg/mL
Sample Type:
Cell culture supernates, serum, and plasma
UniProt:
P16599
Storage:
2-8°C for 12 months.
Stability:
When the kit was stored at the recommended temperature for 12 months, the signal intensity decreased by less than 30%. For unopened kits, if you want to prolong the storage time, please store the Standard, Detection A, Detection B and Microplate at - 20 °C, the rest reagents should be store at 4°C.
Note:
For Research Use Only. Not for diagnostic procedures.
Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
Step
Procedure
1
Preparation: Bring all reagents and samples to room temperature and prepare reagents, working standards and samples as directed. Assay all standards and samples in duplicate.
2
Standard & Sample Incubation: Add 100 µL standards or samples to each well; incubate for 1 hour at 37°C. Wash 3 times with Wash Buffer (300 µL).
3
Detection: Add 100 µL of Detection A working solution to each well; incubate for 1 hour at 37°C. Add 100 µL of Detection B working solution to each well; incubate for 30 minutes at 37°C. Wash 5 times with Wash Buffer (300 µL).
4
Color Development: Add 100 µL of Color Reagent to each well; incubate for 15 minutes at 37°C; protect from light.
5
Stop & Read: Add 50 µL of Stop Solution to each well, then read the optical density at 450 nm within 10 minutes (correction 630 nm or 620 nm).
Sample Type
Protocol
Serum
Use a serum separator tube (SST) and allow samples to clot for 30 minutes at room temperature before centrifugation for 15 minutes at 1000 x g. Remove serum and assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles.
Plasma
Collect plasma using EDTA, heparin, or citrate as an anticoagulant. Centrifuge for 15 minutes at 1000 x g within 30 minutes of collection. Assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles.
Other Sample Types
For more information about how to process other sample types, (e.g., body fluids, breast milk & more), please contact our Tech Support Team at techsupport@assaygenie.com.
