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Human Total P53(Tumor Protein p53) ELISA Kit

This kit is based on Double antibody-Sandwich ELISA detection method and takes 4h assay time. The microplate provided in this kit has been precoated with anti TP53 antibody. Add standard and properly diluted sample into relevant well respectively. After incubation, wash unbound components. Add biotinylated detection antibody. Then, it binds with TP53 bound to precoated antibody. Wash unbound components and add HRP-Streptavidin Conjugate (SABC). Wash unbound components again 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 TP53 in the sample by plotting standard curve. The concentration of the target substance is proportional to the OD450 value.

Alias:TP53 ELISA Kit, BCC7 ELISA Kit, LFS1 ELISA Kit, TRP53 ELISA Kit, Antigen NY-CO-13 ELISA Kit, FLJ92943 ELISA Kit, LFS1TRP53 ELISA Kit, p53 ELISA Kit, p53 tumor suppressor ELISA Kit, P53cellular tumor antigen p53 ELISA Kit, Phosphoprotein p53 ELISA Kit, transformation-related protein 53 ELISA Kit, tumor protein p53 ELISA Kit, Tumor suppressor p53 ELISA Kit
Catalogue No.:EH3898Species:Human
Range:78.125-5000pg/mlSensitivity:46.875pg/ml
  • SPECIFICATIONS
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Product Name
Human Total P53(Tumor Protein p53) ELISA Kit
Alias
TP53 ELISA Kit, BCC7 ELISA Kit, LFS1 ELISA Kit, TRP53 ELISA Kit, Antigen NY-CO-13 ELISA Kit, FLJ92943 ELISA Kit, LFS1TRP53 ELISA Kit, p53 ELISA Kit, p53 tumor suppressor ELISA Kit, P53cellular tumor antigen p53 ELISA Kit, Phosphoprotein p53 ELISA Kit, transformation-related protein 53 ELISA Kit, tumor protein p53 ELISA Kit, Tumor suppressor p53 ELISA Kit
Catalogue No.
EH3898
Size
48T/96T
Species
Human
UniProt No.
P04637
Sample Type
Serum, Plasma, Cell Culture Supernatant, cell or tissue lysate, Other liquid samples
Detection Method
Sandwich ELISA, Double Antibody
Detection Wavelength
OD450
Reaction Duration
4 hours
Range
78.125-5000pg/ml
Sensitivity
46.875pg/ml
Storage
2-8°C(Sealed), Don't cryopreserve.
Specificity
Specifically binds with TP53 , 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 Biotin-labeled Antibody(Concentrated, 100X) 60ul 120ul 2-8°C (Avoid Direct Light)
E034 HRP-Streptavidin Conjugate(SABC, 100X) 60ul 120ul
E024 TMB Substrate 5ml 10ml
E039 Sample Dilution Buffer 10ml 20ml 2-8°C
E040 Antibody Dilution Buffer 5ml 10ml
E049 SABC 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
  • Step 1: Add 100ul standard or sample into each well, seal the plate and statically incubate for 90 minutes at 37°C.
  • Washing: Wash the plate twice without immersing.
  • Step 2: Add 100ul biotin-antibody working solution, seal the plate and statically incubate for 60 minutes at 37°C.
  • Washing: Wash the plate three times and immerse for 1min each time.
  • Step 3: Add 100ul HRP-Streptavidin Conjugate (SABC) working solution, seal the plate and statically incubate for 30 minutes at 37°C.
  • Washing: Wash the plate five times and immerse for 1min each time.
  • Step 4: Add 90ul TMB substrate solution, seal the plate and statically incubate for 10-20 minutes at 37°C. (Accurate TMB visualization control is required.)
  • Step 5: Add 50ul stop solution. Read at 450nm immediately and calculate.
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.(pg/ml) OD-1 OD-2 Average
0 0.081 0.085 0.083
78.125 0.158 0.166 0.161
156.25 0.252 0.265 0.257
312.5 0.458 0.481 0.467
625 0.726 0.763 0.741
1250 1.149 1.207 1.172
2500 1.661 1.746 1.695
5000 2.072 2.177 2.114
Recovery

Add a certain amount of TP53 into the sample. Calculate the recovery by comparing the measured value with the expected amount of TP53 in the sample.

Sample Type Recovery Range(%) Average(%)
serum(n=10) 98-103 93
EDTA plasma(n=10) 91-101 96
Heparin plasma(n=10) 87-103 93
Linearity

Dilute the sample with a certain amount of TP53 at 1:2, 1:4 and 1:8 to get the recovery range.

Sample Type 1:2 1:4 1:8
serum(n=10) 85-88% 92-98% 85-104%
EDTA plasma(n=10) 89-100% 89-99% 82-101%
Heparin plasma(n=10) 83-97% 81-99% 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 (pg/ml) 155.5 610.2 2535 159.3 632.2 2526
Standard deviation 8.09 30.45 128.27 7.87 30.85 130.34
CV(%) 5.2 4.99 5.06 4.94 4.88 5.16
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
Journal:
Biomedical and Pharmacology Journal
Author:
Centre for Toxicology and Health Risk Studies (CORE), Faculty of Health Sciences, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, Kuala Lumpur, Malaysia.
Cited Date:
2023-10-13
Product: