Hamster Tumor Necrosis Factor (TNF) ELISA Kit, Lot 21AO-159 [Cancer Immune Checkpoint Assay Kit] (CAT#: IOK-05-P070)

Pre-coated, ready to use 96-well strip plate
Plate sealer for 96 wells
Standard
Sample/Standard Dilution Buffer
Biotin-labeled Antibody (Concentrated)
Antibody Dilution Buffer
HRP-Streptavidin Conjugate (SABC)
SABC Dilution Buffer
TMB Substrate
Stop Solution
Wash Buffer (25 x concentrate)
Instruction manual

1.Microplate reader (wavelength:450nm)
2.37 °C incubator
3.Automated plate washer
4.Precision single and multi-channel pipette and disposable tips
5.Clean tubes
6.Deionized or distilled water

1.875 pg/mL

100 μL

Pre-coated

Bring all reagents and samples to room temperature for 20 minutes before use.
Wash Buffer
Standards
Preparation of Biotin-labeled Antibody Working Solution
Preparation of HRP-Streptavidin Conjugate (SABC) Working Solution

Set standard, test samples (diluted at least 1/2 with Sample Dilution Buffer), control (blank) wells on the pre-coated plate respectively, and then, records their positions. Wash plate 2 times before adding standard, sample and control (blank) wells.
Prepare Standards
Add Samples
Incubate
Wash
Biotin-labeled Antibody
Wash
HRP-Streptavidin Conjugate (SABC)
Wash
TMB Substrate
Stop
OD Measurement

Intra-Assay: CV<8%
Inter-Assay: CV<10%

4 °C,-20 °C

1.For unopened kit: All the reagents should be kept according to the labels on vials. The Reference Standard, Biotin-labeled Antibody, HRP Conjugate and the 96-well stripe plate should be stored at -20 °C upon receipt while the other reagents should be stored at 4 °C.
2.For used kit: When the kit is used, the remaining reagents need to be stored according to the above storage condition. Besides, please return the unused wells to the foil pouch containing the desiccant pack, and zip-seal the foil pouch.

6 months

1.For unopened kit: All the reagents should be kept according to the labels on vials. The Reference Standard, Biotin-labeled Antibody, HRP Conjugate and the 96-well stripe plate should be stored at -20 °C upon receipt while the other reagents should be stored at 4 °C.
2.For used kit: When the kit is used, the remaining reagents need to be stored according to the above storage condition. Besides, please return the unused wells to the foil pouch containing the desiccant pack, and zip-seal the foil pouch.

For Research Use only

Tumor Necrosis Factor Alpha (TNFa)

DIF; TNF-alpha; TNFA; TNFSF2; RATTNF; Tnfa; tnf; TNF-a; TNFalpha; Tnfsf1a; TNFa; cTNF; Tnf-alpha; tnfa-like; TNF-ALPHA; dif; tnfa; xtnf; tnfsf2; tnf-alpha; Cachectin; tumor necrosis factor; tumor necrosis factor b (TNF superfamily; member 2); tumor necrosis factor alpha; tumor necrosis factor a (TNF superfamily; member 2); TNF; Tnf; tnf; tnfb; tnf-alpha; LOC103694380; tnfa

TNF-α, DIF, TNF-alpha, TNFA, TNFSF2

A0A1U7RE37

NF-kappaB Signaling, Apoptosis, Caspase Cascade in Apoptosis, TLR Signaling, Cellular Response to Molecule of Bacterial Origin, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response, Positive Regulation of Endopeptidase Activity, Hepatitis C, Protein targeting to Nucleus, Inflammasome

1.Wash plate 2 times before adding Standard, Sample and Control wells!
2.Add 100 µL standard or sample to each well and incubate for 90 minutes at 37 °C.
3.Aspirate and wash plates 2 times.
4.Add 100 µL Biotin-labeled antibody working solution to each well and incubate for 60 minutes at 37 °C.
5.Aspirate and wash plates 3 times.
6.Add 100 µL SABC Working Solution into each well and incubate for 30 minutes at 37 °C.
7.Aspirate and wash plates 5 times.
8.Add 90 µL TMB Substrate Solution. Incubate 10-20 minutes at 37 °C.
9.Add 50 µL Stop Solution. Read at 450nm immediately and calculation.