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

Citations(111)Uniprot : P01375

Cat.No.:E-EL-H0109
Reactivity: Human

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	7.81-500 pg/mL
Sensitivity	4.69 pg/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Human TNF-α in samples. No significant cross-reactivity or interference between Human TNF-α and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 10%.
Application	This ELISA kit applies to the in vitro quantitative determination of Human TNF-α concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human TNF-α. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human TNF-α and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human TNF-α, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human TNF-α. You can calculate the concentration of Human TNF-α in the samples by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	-20℃, 6 months
Reference Standard	96T: 2 vials 48T: 1 vial
Concentrated Biotinylated Detection Ab (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 6 months
Biotinylated Detection Ab Diluent	1 vial, 14 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Substrate Reagent	1 vial, 10 mL	2-8℃(Protect from light)
Stop Solution	1 vial, 10 mL	2-8°C
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human TNF-α were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human TNF-α were tested on 3 different plates, 20 replicates in each plate.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (pg/mL)	24.10	79.50	183.00	22.70	86.50	177.40
Standard deviation	1.50	3.80	9.70	1.20	4.00	7.60
CV (%)	6.22	4.78	5.30	5.29	4.62	4.28

Recovery

The recovery of Human TNF-α spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type	Range (%)	Average Recovery (%)
Serum(n=8)	86-100	92
EDTA plasma (n=8)	94-106	99
Cell culture media (n=8)	93-107	99

Linearity

Samples were spiked with high concentrations of Human TNF-α and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database Links	SwissProt: P01375
Synonyms	DIF, TNF-alpha, TNFA, TNFSF2
Research Area	Cancer,Immunology,Signaling transduction,Stem cells

Assay Procedures

	1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
	2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
	3. Aspirate and wash the plate for 3 times
	4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	6. Add 50μL Stop Solution
	7. Read the plate at 450nm immediately. Calculation of the results

Citations

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DOI: 10.2147/IJN.S134897
PMID: 28652743	Sample: Cell culture supernatant,Serum

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DOI: 10.1155/2017/8519169
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DOI: 10.1038/srep15132
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DOI: 10.1021/acs.bioconjchem.6b00600
PMID: 28004910	Sample: Cell culture supernatant

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DOI: 10.3389/fcimb.2016.00109
PMID: 27709104	Sample: Cell culture supernatant

Scientific reports (2020) IF: 3.998

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DOI: 10.1038/s41598-020-68543-9
PMID: 32669704	Sample: Cell culture supernatant

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DOI: 10.1016/j.fsi.2020.06.024
PMID: 32585358	Sample: Cell culture supernatant

European Journal of Pharmacology (2020) IF: 3.263

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DOI: 10.1016/j.yexcr.2020.112086
PMID: 32416091	Sample: Serum,Cell sample

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DOI: 10.1016/j.jep.2020.113039
PMID: 32497675	Sample: Serum,Cell culture supernatant

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DOI: 10.26355/eurrev_202006_21521
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DOI: 10.1016/j.jep.2019.112433
PMID: 31783135	Sample: Cell culture supernatant

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DOI: 10.26355/eurrev_201910_19167
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DOI: 10.1007/s10495-018-1474-7
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DOI: 10.1016/j.lfs.2018.12.020
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DOI: 10.1016/j.molimm.2018.12.027
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DOI: 10.1016/j.biopha.2019.108706
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DOI: 10.1016/j.biopha.2018.06.044
PMID: 29966972	Sample: Serum

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DOI: 10.1155/2017/3605350
PMID: 29249869	Sample: Serum

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DOI: 10.3390/ijms18081614
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DOI: 10.1007/s10495-015-1162-9
PMID: 26283170	Sample: Cell culture supernatant

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DOI: 10.3748/wjg.v22.i17.4354
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DOI: 10.1080/13813455.2020.1789880
PMID: 32669002	Sample: Serum

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DOI: 10.1631/jzus.B2000174
PMID: 32425003	Sample: Serum,Plasma

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DOI: 10.1007/s12011-019-01831-2
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DOI: 10.1016/j.bbrc.2019.10.135
PMID: 31668807	Sample: Cell culture supernatant

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DOI: 10.1111/sji.12868
PMID: 32052490	Sample: Serum

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DOI: 10.1093/gastro/goz058
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DOI: 10.1016/j.psj.2019.10.008
PMID: 32036960	Sample: Plasma

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DOI: 10.1007/s12640-020-00193-1
PMID: 32335809	Sample: Serum

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DOI: 10.1016/j.micpath.2019.103719
PMID: 31493501	Sample: Cell culture supernatant

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DOI: 10.1111/apm.12925
PMID: 30746790	Sample: Cell culture supernatant

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DOI: 10.1155/2019/5961742
PMID: 30941196	Sample: Cell culture supernatant

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DOI: 10.1016/j.biopha.2017.11.151
PMID: 29793333	Sample: Cell culture supernatant,Serum

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DOI: 10.1371/journal.pone.0181440
PMID: 28837559	Sample: Serum

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DOI: 10.1016/j.carpath.2017.03.009
PMID: 28433861	Sample: Cell culture supernatant

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DOI: 10.3390/molecules201219764
PMID: 26633327	Sample: Cell lysate

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DOI: 10.1016/j.tiv.2016.05.017
PMID: 27247145	Sample: Cell culture supernatant

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DOI: 10.1016/j.ijid.2015.12.009
PMID: 26701820	Sample: Cell culture supernatant

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DOI: 10.1371/journal.pone.0068963
PMID: 23894384	Sample: Plasma

Evidence-Based Complementary and Alternative Medicine (2020) IF: 1.813

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DOI: 10.1155/2020/5834502
PMID: 32714409	Sample: Cell culture supernatant

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DOI: 10.1016/j.archoralbio.2020.104858
PMID: 32805637	Sample: Cell culture supernatant

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DOI: 10.3892/etm.2020.8903
PMID: 32782497	Sample: Cell culture supernatant

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Correlation Between Single Nucleotide Polymorphisms at the 3'-UTR of the NFKB1 Gene and Acute Kidney Injury in Sepsis

DOI: 10.1089/gtmb.2019.0222
PMID: 32315555	Sample: Cell sample

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Avicularin Reduces the Expression of Mediators of Inflammation and Oxidative Stress in Bradykinin-Treated MG-63 Human Osteoblastic Osteosarcoma Cells

DOI: 10.12659/MSM.921957
PMID: 32463805	Sample: Cell culture supernatant

Experimental and Therapeutic Medicine (2020) IF: 1.785

Systematically transplanted human gingiva-derived mesenchymal stem cells regulate lipid metabolism and inflammation in hyperlipidemic mice with periodontitis

DOI: 10.3892/etm.2019.8256
PMID: 31885706	Sample: Serum

ANZ Journal of Surgery (2020) IF: 1.355

Effect of laparoscopic sleeve gastrectomy on renal function in obese patients

DOI: 10.1111/ans.15767
PMID: 32115876	Sample: Serum,Urine

International Journal of Environmental Analytical Chemistry (2020) IF: 1.431

A label-free and sensitive impedimetric immunosensor for TNF-α biomarker detection based on epoxysilane-modified disposable ITO-PET electrode

DOI: 10.1080/03067319.2019.1679807
PMID: 99999999	Sample: Immunosensor Construction

medicine (2019) IF: 1.552

A case-control study on the relationship between miR-As single nucleotide polymorphisms and sepsis risk

DOI: 10.1097/MD.0000000000016744
PMID: 31415369	Sample: Serum

Genetic testing and molecular biomarkers (2018) IF: 1.181

Association of High Mobility Group Box Protein B1 Gene Polymorphisms with Pneumonia Susceptibility and Severity

DOI: 10.1089/gtmb.2018.0174
PMID: 30562142	Sample: Plasma

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Mitochondrial DNA-Induced Inflammatory Responses and Lung Injury in Thermal Injury Murine Model: Protective Effect of Cyclosporine-A

DOI: 10.1093/jbcr/irz029
PMID: 30926991	Sample: Serum

Experimental and therapeutic medicine (2018) IF: 1.41

Anti-inflammatory Effect of Resolvin D1 on LPS-treated MG-63 Cells

DOI: 10.3892/etm.2018.6721
PMID: 30402165	Sample: Serum

Experimental and Therapeutic Medicine (2019) IF: 1.41

Effects of propofol combined with remifentanil anesthesia on the NO, endothelin and inflammatory cytokines in the plasma of patients with liver cirrhosis during the perioperative period

DOI: 10.3892/etm.2019.7367
PMID: 30988754	Sample: Plasma

Molecular Medicine Reports (2017) IF: 1.692

Tetrahydroxy Stilbene Glucoside Ameliorates H2O2-Induced Human Brain Microvascular Endothelial Cell Dysfunction In Vitro By Inhibiting Oxidative Stress And Inflammatory Responses.

DOI: 10.3892/mmr.2017.7225
PMID: 28849141	Sample: Cell culture supernatant

Molecular Medicine Reports (2018) IF: 1.692

The Shortening Telomere Length Of T Lymphocytes Maybe Associated With HyperFunction In Servere Aplastic Anemia.

DOI: 10.3892/mmr.2017.8014
PMID: 29115638

Experimental and Therapeutic Medicine (2018) IF: 1.261

Role Of miR-589-3p In Human Lumbar Disc Degeneration And Its Potential Mechanism.

DOI: 10.3892/etm.2017.5593
PMID: 29434746	Sample: Cell culture supernatant

Experimental and Therapeutic Medicine (2017) IF: 1.261

Inhibition Of miR27a Suppresses The Inflammatory Response Via The p38/MAPK Pathway In Intervertebral Disc Cells.

DOI: 10.3892/etm.2017.5053
PMID: 29067127	Sample: Cell culture supernatant

The Journal of Gene Medicine (2020)

Human cytomegalovirus infection is correlated with atherosclerotic plaque vulnerability in carotid artery

DOI: 10.1002/jgm.3236
PMID: 32468600	Sample: Serum

Int J Curr Microbiol App Sci (2015)

The role of TNF and Resistin Gene+ 299 (G/A) polymorphism in the development of insulin resistance in non-obese Type 2 Diabetes Mellitus Iraqi patients


PMID: 99999999	Sample: Serum

Biomedical and Pharmacology Journal (2016)

Microbial Index and TNF-α, IL-4, CCL17 Level Among Burn Wound Infections in Hilla City, Iraq.

DOI: 10.13005/bpj/1054
PMID: 99999999	Sample: Serum

European Microscopy Congress 2016: Proceedings (2016)

The activation, ROS induction and cytokine secretion in human monocytes exposed to H . pylori lipopolysaccharide depend on the availability of TLR4/2 and TLR2/6 receptor complexes

DOI: 10.1002/9783527808465.EMC2016.6255
PMID: 99999999	Sample: Cell culture supernatant

JMSCR (2018)

Evaluation of Cytokines Response in Plasmodium falciparum Infection among Adolescents in Port Harcourt, Nigeria

DOI: 10.18535/jmscr/v6i9.54
PMID: 99999999	Sample: Plasma

Preprint (2019)

The neuro-immune fingerprint of major neuro-cognitive psychosis or deficit schizophrenia: a supervised machine learning study

DOI: 10.20944/preprints201905.0285.v1
PMID: 99999999	Sample: Serum

Indian Journal of Forensic Medicine & Toxicology (2019)

Influence of TLR4 Polymorphisms on Susceptibility to UTI Caused by Gram Negative Bacteria in Pregnant Women

DOI: 10.5958/0973-9130.2019.00454.7
PMID: 99999999	Sample: Serum,Urine

The Indonesian Biomedical Journal (2019)

TNFR, TRAF2, NF-κB mRNA Levels of Glioblastoma Multiforme Cells Treated by Conditioned Medium of Umbilical Cord-derived Mesenchymal Stem Cells

DOI: 10.18585/inabj.v11i2.722
PMID: 99999999	Sample: Cell lysate

International Journal of Tropical Disease & Health (2019)

Correlation of Small Dense Low Density Lipoprotein, Tumour Necrosis Factor-alpha with Liver Enzymes in Chronic Hepatitis B Patients

DOI: 10.9734/ijtdh/2019/v37i430172
PMID: 99999999	Sample: Serum

Indian Journal of Public Health (2020)

Association of TLR4 Polymorphisms with Susceptibility to Urinary Tract Infection Caused by Gram Positive Bacteria in Pregnant Women


PMID: 99999999	Sample: Serum

The Journal of Research on the Lepidoptera (2020)

Estimation of IL-1βand TNF Concentration in Children Suffering from Febrile Convulsion

DOI: 10.36872/LEPI/V51I2/301090
PMID: 99999999	Sample: Serum

Preprint (2020)

Thymosin β10 promotes tumor-associated macrophages M2 conversion and proliferation via the PI3K/Akt pathway in lung adenocarcinoma

DOI: 10.21203/rs.3.rs-34263/v1
PMID: 99999999	Sample: Cell culture supernatant

The Journal of Immunology (2020)

CALML6 Controls TAK1 Ubiquitination and Confers Protection against Acute Inflammation

DOI: 10.4049/jimmunol.1901042
PMID: 32303555	Sample: Cell culture supernatant

International Journal of Drug Delivery Technology (2020)

Evaluation of Synergistic Effect of Cytosine-PhosphodiesterbondGuanosin-Oligodeoxynucleotide 7909, and Protamine on Transfection Process Mediated by Calcium Phosphate Nanoparticles


PMID: 99999999	Sample: Cell culture supernatant

Preprints (2020)

Lowered serum cesium levels in schizophrenia: association with immune-inflammatory biomarkers and cognitive impairments

DOI: 10.1590/1516-4446-2020-0908
PMID: 32556004	Sample: Serum

Annals of African Medicine (2019)

Hyperhomocysteinemia and Folate Levels in Normal Healthy Nigerians Living in Zaria: Subanalysis of ABU Homocysteine Cross-sectional Survey

DOI: 10.4103/aam.aam_53_18
PMID: 31417015	Sample: Plasma

Revista da Associa??o Médica Brasileira (2019)

TLR7 mediates increased vulnerability to ischemic acute kidney injury in diabetes

DOI: 10.1590/1806-9282.65.8.1067
PMID: 31531603	Sample: Cell culture supernatant

The Annals of thoracic surgery (2019)

Impact of an Early Oral Feeding Protocol on Inflammatory Cytokine Changes After Esophagectomy

DOI: 10.1016/j.athoracsur.2018.09.048
PMID: 30403976	Sample: Serum

Modern Research in Inflammation (2019)

Estimation of Cytokines Involved in Acute-Phase Wound Infection with Reference to Residence Time of Patients in Hospitals

DOI: 10.4236/mri.2019.81001
PMID: 99999999	Sample: Serum

NTERNATIONAL JOURNAL OF CLINICAL AND EXPERIMENTAL MEDICINE (2019) IF: 0.181

Resveratrol protects human renal proximal tubular cells from high glucose and hypoxia/reoxygenation induced injury via inhibiting p38-MAPK and thioredoxin-interacting protein (TXNIP) pathways


PMID: 99999999	Sample: Cell culture supernatant

Journal of Pharmaceutical Sciences and Research (2018)

Effect of General and Spinal Anesthesia on Some Anti-inflammatory cytokines and cell adhesion molecules of Iraqi Women Undergo Cesarean Section: Comparative Study


PMID: 99999999	Sample: Serum

Medical science monitor: international medical journal of experimental and clinical research (2018)

Sinomenine Regulates Inflammatory Response and Oxidative Stress via Nuclear Factor kappa B (NF-kB) and NF-E2-Related Factor 2 (Nrf2) Signaling Pathways in Ankle Fractures in Children

DOI: 10.12659/MSM.910740
PMID: 30237391	Sample: Serum

University of Sulaimani (2018)

EFFECTS OF GINKGO BILOBA EXTRACT (EGb761) AS ADDON WITH METFORMIN IN THE TREATMENT OF PATIENTS


PMID: 99999999	Sample: Serum

Medical Journal of Babylon (2017)

Tumor Necrosis Factor Alpha Synergetic with Insulin Resistance Potentially Contributes to the Development of Polycystic Ovary Syndrome.


	Sample: Serum

ALMUSTANSIRIYAH Journal of PHARMACEUTICAL SCIENCES (AJPS) (2018)

Potential Benefit of Melatonin as an Adjuvant Therapy in a Sample of Iraqi Patients with Crohn’s Disease: A New Study.


	Sample: Serum

Kerbala Jorunal of Medicine (2017)

Serum Tnfα Levels Are Inversely Associated With Insulin Sensitivity in Diabetic Retinopathy Patients.


	Sample: Serum

Medical Science Monitor: International Medical Journal of Experimental And Clinical Research (2018) IF: /

Propofol Promotes Ankle Fracture Healing in Children by Inhibiting Inflammatory Response.

DOI: 10.12659/MSM.908592
PMID: 29940605	Sample: Serum

Pediatric Allergy (2018) IF: 0.533

High-Mobility Group Protein Box 1 is Upregulated in Children with Henoch-Schonlein Purpura.

DOI: 10.1089/ped.2018.0872
	Sample: Serum

Experimental and Therapeutic Medicine (2018)

Leukocyte Depletion Does Not Affect The In Vitro Healing Ability Of Platelet Rich Plasma.

DOI: 10.3892/etm.2018.5887
PMID: 29556269

Iraq Medical Journal (2018)

Adiponectin may be used as a marker in prediction of diabetic retinopathy patients.


	Sample: Serum

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Reviews/Q&A

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Q&A

S*********nSubmitted [ Aug 05 2021 ]

Asked: My kit got expired in April 2020. Shall we perform the experiment with the kit?

adminSubmitted [ Aug 05 2021 ]

Answered: Thanks for your message. The expired kit can be used to perform experiment anymore. By the way, where did you ordered the kit from?

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