Human anti-TNF-α Antibody(Anti Tumor Necrosis Factor Alpha Antibody) ELISA Kit

Category:

This kit is based on indirect ELISA detection method and takes 3h assay time. The microplate provided in this kit has been precoated with antigen. HRP-antibody is used as detection antibody. Add standard and properly diluted sample into relevant well respectively. After incubation, wash unbound components. Add HRP-detection antibody, then it binds with anti-TNF-α Antibody bound to precoated antibody. Wash unbound components and add TMB substrate solution. Then, TMB was catalyzed by HRP to produce a blue color product that turned yellow after adding a stop solution. Read the O.D. absorbance at 450nm in a microplate reader. Calculate the concentration of anti-TNF-α Antibody in the sample by plotting standard curve. The concentration of the target substance is proportional to the OD450 value.

Catalogue No.: 
EH4080
Alias: 
anti-TNF-α Antibody ELISA Kit, Anti Tumor Necrosis Factor Alpha Antibody ELISA Kit
Species: 
Human
Range: 
1.563-100ng/ml
Sensitivity: 
0.938ng/ml
  • SPECIFICATIONS
Product Name
Human anti-TNF-α Antibody(Anti Tumor Necrosis Factor Alpha Antibody) ELISA Kit
Alias
anti-TNF-α Antibody ELISA Kit, Anti Tumor Necrosis Factor Alpha Antibody ELISA Kit
Catalogue No.
EH4080
Size
48T/96T
Species
Human
UniProt No.
P01375
Sample Type
Serum, Plasma, Cell Culture Supernatant, cell or tissue lysate, Other liquid samples
Detection Method
HRP Indirect ELISA Sandwich
Detection Wavelength
OD450
Reaction Duration
3 hours
Range
1.563-100ng/ml
Sensitivity
0.938ng/ml
Storage
2-8°C(Sealed), Don't cryopreserve.
Specificity
Specifically binds with anti-TNF-α Antibody , no obvious cross reaction with other analogues.
ELISA Kit Components
Kit Components Item Size(48T) Size(96T) Storage Condition for Opened Kit
E001 ELISA Microplate(Dismountable) 8×6 8×12 Put the rest strips into a sealed foil bag with the desiccant. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C
E002 Lyophilized Standard 1vial 2vial Put the rest standards into a desiccant bag. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C
E003 HRP Antibody(Concentrated, 100X) 60ul 120ul 2-8°C (Avoid Direct Light)
E024 TMB Substrate 5ml 10ml
E039 Sample Dilution Buffer 10ml 20ml 2-8°C
E040 Antibody Dilution Buffer 5ml 10ml
E026 Stop Solution 5ml 10ml
E038 Wash Buffer(Concentrated, 25X) 15ml 30ml
E006 Plate Sealer 3 pieces 5 pieces  
E007 Product Description 1 copy 1 copy
Required Instruments and Reagents
  1. Microplate reader (wavelength: 450nm)
  2. 37°C incubator (CO2 incubator for cell culture is not recommenced.)
  3. Automated plate washer or multi-channel pipette/5ml pipettor (for manual washing purpose)
  4. Precision single (0.5-10μL, 5-50μL, 20-200μL, 200-1000μL) and multi-channel pipette with disposable tips(Calibration is required before use.)
  5. Sterile tubes and Eppendorf tubes with disposable tips
  6. Absorbent paper and loading slot
  7. Deionized or distilled water
Assay Procedure Summary
Elisa实验原理图
Standard Curve

This product is detected by QC department and meets performance required in the manual. (Laboratory Humidity: 20%-60%; Temperature: 18°C -25°C; Equilibrate TMB substrate to 37°C before staining. After adding into the ELISA wells, incubate for 15min at 37°C in dark.)

Due to different assay environments and operations, assay data below and standard curve are provided for reference. Experimenters should establish standard curve according to their own assay.

STD.(ng/ml) OD-1 OD-2 Average
0 0.043 0.045 0.044
1.562 0.094 0.098 0.096
3.125 0.164 0.17 0.167
6.25 0.217 0.225 0.221
12.5 0.357 0.371 0.364
25 0.624 0.65 0.637
50 1.229 1.279 1.254
100 2.113 2.199 2.156
EH4080 Standard Curve Image
Recovery

Add a certain amount of anti-TNF-α Antibody into the sample. Calculate the recovery by comparing the measured value with the expected amount of anti-TNF-α Antibody in the sample.

Sample Type Recovery Range(%) Average(%)
serum(n=10) 91-104 97
EDTA plasma(n=10) 90-104 100
Heparin plasma(n=10) 86-99 92
Linearity

Dilute the sample with a certain amount of anti-TNF-α Antibody at 1:2, 1:4 and 1:8 to get the recovery range.

Sample Type 1:2 1:4 1:8
serum(n=10) 89-104% 85-93% 80-91%
EDTA plasma(n=10) 88-105% 85-99% 91-97%
Heparin plasma(n=10) 88-101% 87-99% 80-97%
Precision(%)

Intra-assay Precision: samples with low, medium and high concentration are tested 20 times on the same plate.

Inter-assay Precision: samples with low, medium and high concentration are tested 20 times on three different plates.

Item Intra-assay Precision Inter-assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (ng/ml) 3.23 12.37 48.35 3.12 12.79 49.66
Standard deviation 0.15 0.69 3.06 0.18 0.75 2.86
CV(%) 4.69 5.54 6.32 5.86 5.85 5.75
Stability

Perform the stability test for the sealed kit at 37°C and 2-8°C and get relevant data.

ELISA kit(n=5) 37°C for 1 month 2-8°C for 6 months
Average(%) 80 95-100