Elisa

Manufactured by PerkinElmer

Sourced in United States

ELISA (Enzyme-Linked Immunosorbent Assay) is a widely used analytical biochemistry assay technique that detects and quantifies specific proteins, hormones, antibodies, and other biomolecules. It utilizes the high specificity of antibody-antigen interactions and employs enzyme-linked detection methods to generate a measurable signal proportional to the concentration of the target analyte in a sample.

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Lab products found in correlation

Penicillin, by Thermo Fisher Scientific (7 mentions) Streptomycin, by Thermo Fisher Scientific (6 mentions) Millex hv, by Merck Group (3 mentions) Calphos, by Takara Bio (3 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Sas 9, by SAS Institute (3 mentions) Dnase rq1 rnase free dnase, by Promega (2 mentions) Cd4 t cell isolation kit 2, by Miltenyi Biotec (2 mentions) Anti cd25, by Miltenyi Biotec (2 mentions) Anti cd69 microbead kit 2, by Miltenyi Biotec (2 mentions) Anti hla dr microbeads, by Miltenyi Biotec (2 mentions) Anti biotin microbead, by Miltenyi Biotec (2 mentions) Fetal bovine serum (fbs), by American Type Culture Collection (2 mentions) Rpmi 1640 media, by Thermo Fisher Scientific (2 mentions) 0.2 m filter, by Avantor (2 mentions) Sctδ19, by Thermo Fisher Scientific (2 mentions) X tremegene hp transfection reagent, by Merck Group (2 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions) Pen strep, by Thermo Fisher Scientific (2 mentions) Miscript system, by Qiagen (1 mentions)

47 protocols using elisa

Molecular Regulation of Insulin Secretion

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Cytokine treatment of cells was performed with 5 ng/ml IL1β from R&D systems. RNA isolation and reverse transcription followed by quantitative real-time PCR (qRT-PCR) were performed (Qiagen miScript system) [6 (link)]. miRNA or mRNA expression was quantified relative to U6 or β-actin, respectively, using the comparative Ct method (Primer sequences in Supplementary Table 1) [6 (link)]. Pulldown of mRNAs bound to 50 nM biotinylated miR-21-3p, miR-21-5p, or control Caenorhabditis elegans miR-67 was performed as described [27 ]. Luciferase assays were performed using a Gaussia luciferase/secreted alkaline phosphatase dual reporter system (GeneCopoeia) and wild-type rat Tgfb2 3′ untranslated region (UTR) and Smad2 3′UTR or mutated 3′UTRs for Tgfb2 (positions 1281–1289) and Smad2 (positions 8900–8908) [6 (link)]. Immunoblotting was performed as described, visualized using an Odyssey imaging system, and quantified by LI-COR software (LI-COR Biotech) (antibodies in Supplementary Table 2) [6 (link)].
Static GSIS and perifusion were performed as described with supernatants assayed for insulin using ELISA (Cisbio) and normalized to total DNA content (PICO Green Assay; Invitrogen) [28 (link), 29 (link)].

Ibrahim S., Johnson M., Stephens C.H., Xu J., Moore R., Mariani A., Contreras C., Syed F., Mirmira R.G., Anderson R.M, & Sims E.K. (2021). β-Cell pre-mir-21 induces dysfunction and loss of cellular identity by targeting transforming growth factor beta 2 (Tgfb2) and Smad family member 2 (Smad2) mRNAs. Molecular Metabolism, 53, 101289.

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Serum Sclerostin Levels in Postmenopausal Breast Cancer

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Blood samples were collected in the morning after an overnight fast. Routine serum chemistry determinations including calcium and phosphate were performed by standard automated techniques. Bone turnover markers were measured using the following methods: osteocalcin (OCN; by enzyme-linked immunosorbent assay [ELISA], CIS Bio International, Gif-sur-Yvette, France; intra-assay coefficient of variation [CV], <2.0%; interassay CV, <5.0%), C-telopeptide of type I collagen (CTx; Osteomark, Ostex International, Seattle, WA, USA; intra-assay CV, <5.8%; interassay CV, <5.9%), intact PTH (by immunoradiometric assay, Biosource, Nivelles Belgium; intra-assay CV, 2.7%; interassay CV, <3.5%), and 25-hydroxyvitamin D (25[OH]D by D3-radioimmunoassay-coated tube, Biosource; intra-assay CV, 11.0%; interassay CV, 12.5%).
Serum sclerostin concentrations were measured using a human sclerostin ELISA kit (Biomedica Co., Wien, Austria) according to the manufacturer's instructions. Intraassay and interassay CVs were 4% to 6% and 5% to 7%, respectively. Sclerostin measurements are reported in pmol/L, and the lower limit of detection was <8.9 pmol/L. Measurements were taken twice at intervals of 24 weeks in postmenopausal women with endocrine-responsive breast cancer. Sclerostin was measured only at baseline in the premenopausal control group.

Kim W., Chung Y., Kim S.H., Park S., Bae J.H., Kim G., Lee S.J., Kim J.E., Park B.W., Lim S.K, & Rhee Y. (2015). Increased Sclerostin Levels after Further Ablation of Remnant Estrogen by Aromatase Inhibitors. Endocrinology and Metabolism, 30(1), 58-64.

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HIV-1 Infection of Human Tissues

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HIV-1 stocks were generated in human PBMCs. Tissues were infected with 10⁴ cell-free R5-tropic HIV-1_BaL, a 50% Tissue Culture Infectious Dose (TCID₅₀)/ml. After overnight incubation at 37°C, tissues were washed to remove residual input virus (day 0), and cultured for up to 21 days in Leibowitz (L)15 medium (GIBCO, Grand Island, NY) as described [7 (link), 17 (link), 18 (link)]. Day’s 11 and 21 supernatants were evaluated for HIV-1 p24 antigen by ELISA (Perkin Elmer, Boston, MA).

Macura S.L., Lathrop M.J., Gui J., Doncel G.F., Asin S.N, & Rollenhagen C. (2016). Blocking CXCL9 Decreases HIV-1 Replication and Enhances the Activity of Prophylactic Antiretrovirals in Human Cervical Tissues. Journal of Acquired Immune Deficiency Syndromes (1999), 71(5), 474-482.

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Cytotoxicity Evaluation of Ara-C and PA-Ara on Leukemia Cell Lines

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On account of the application in the treatment of leukemia, the cytotoxicity of Ara-C and PA-Ara prodrug were evaluated on HL60 and K562 cell lines with an MTT assay and were cultured in IMDM and RPMI-1640 medium, respectively both containing 10% fetal bovine serum (GIBCO, Vitrogen Corporation) and antibiotics (100 units mL⁻¹ penicillin, and 100 μg mL⁻¹ streptomycin) at 37 °C under a humidified atmosphere containing 5% CO₂.^{32 (link)} Exponentially growing HL60 and K562 in 100 μL of culture medium containing different concentrations of samples were seeded into 96-well plates at a density of 3 × 10⁴ and 1.5 × 10⁴ cells per well, respectively. After 24 h or 48 h of incubation at 37 °C, 10 μL of 5% MTT regent was added to each well and incubated at 37 °C for another 4 h. Then the solution containing unreacted MTT was removed carefully and 150 μL of DMSO was added to dissolve the newly generated blue formazan crystals. The optical density of each well was measured using a microplate reader (ELISA, PerkinElmer) at a wavelength of 490 nm, and the cell inhibition ratio of each sample was calculated according to the following equation:where A_{positive control} and A_black refer to the absorbance of the culture without drug treatment and the culture without cells or drug, respectively, whereas the other treatments were the same as for the samples.

Zhang J., Zhang D., Hu X., Liu R., Li Z, & Luan Y. (2018). Rational design of a new cytarabine-based prodrug for highly efficient oral delivery of cytarabine. RSC Advances, 8(24), 13103-13111.

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Cytarabine Nanoparticle Development and Evaluation

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Cytarabine (Ara-C) was purchased from the Aladdin Industrial Corporation. Palmitic acid (PA), sodium hydroxide, potassium dihydrogen phosphate, sodium dihydrogen phosphate and disodium hydrogen phosphate were bought from Sinopharm Chemical Reagent Co., Ltd. Ethyl chloroformate (EtOCOCl) was obtained from Chengdu Beisite Reagent Co., Ltd. Pepsin (1 : 3000) and trypsin (1 : 250) were purchased from Amresco and Sangon Biotech (Shanghai) Co., Ltd, respectively. Acyclovir (HPLC > 98%) was purchased from Dalian Meilun Biotech Co., Ltd. HL60 and K562 cells were kindly provided by the Immunopharmacology Institute of Shandong University and the Shandong Analysis and Test Center. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) was purchased from Beijing Solarbio Technology Co., Ltd.
The spectroscopy equipment was: an NMR spectrometer (¹H-NMR, Bruker Avance 400), an electrospray tandem mass spectrometer (MS, AB SCIEX API 4000), an FTIR spectrometer (Nicolet 6700), a transmission electron microscope (TEM, JEM-200CX), a high performance liquid chromatograph (HPLC, Agilent Technologies 1200 Series), an HPLC-MS (Agilent 1260 triple quadrupole mass spectrometer equipped with an ESI source), and a microplate reader (ELISA, PerkinElmer).

Zhang J., Zhang D., Hu X., Liu R., Li Z, & Luan Y. (2018). Rational design of a new cytarabine-based prodrug for highly efficient oral delivery of cytarabine. RSC Advances, 8(24), 13103-13111.

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HIV-1 Inhibition via CRISPR-Cas9 and TNFα

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Cas9-Vp160-encoding plasmid pAC154-160Dual (Jaenisch lab, purchased from Addgene, #48240) with or without HIV-1-specific gRNAs T5 and T6 (sequences from^{[1 (link)]}, or a GFP-containing DNA vector were electroporated into U1 cells (AIDS Reagent) using a Bio-Rad Gene Pulser II. HIV-1 p24 production was measured by ELISA (PerkinElmer). Results were normalized by viable cell count (Muse, Merck Millipore), although introduction of plasmid did not have a significant effect on viability compared with mock electroporation. Human recombinant tumor necrosis factor-alpha (TNFalpha, Sigma Aldrich) was introduced at 10 ng/mL.
Statistical analysis was done using Prism version 7 for Mac OS X (GraphPad Software, Inc.). One-way ANOVA was performed followed by Dunnett’s multiple comparisons test for individual comparisons (Supplementary data).

Kim V., Mears B.M., Powell B.H, & Witwer K.W. (2017). Mutant Cas9-transcriptional activator activates HIV-1 in U1 cells in the presence and absence of LTR-specific guide RNAs. Matters, 2017, 10.19185/matters.201611000027.

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Quantification of HIV-1 Vif-Mediated A3G Restriction

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HEK293T cells were seeded in 10 cm tissue culture plates and co-transfected with 10 μg pNL4.3/ΔVif and 0.25 to 10 μg pCMV4_A3G_HA wild type or mutant expression vectors using polyethylenimine (PEI, Molecular Biosciences). Total DNA levels were kept constant with empty pCMV4-HA vector. At 48 h, supernatants were harvested, DNAse (RQ1 RNase free DNAse (Promega)) treated for 1 h at 37°C and viruses then purified through a 20% (w/v) sucrose cushion at 28,000 x g for 75 min at 4°C. Viruses were resuspended in PBS, and quantified according to p24^Gag content using an enzyme-linked immunosorbent assay (ELISA; Perkin-Elmer).

Pollpeter D., Parsons M., Sobala A.E., Coxhead S., Lang R.D., Bruns A.M., Papaioannou S., McDonnell J.M., Apolonia L., Chowdhury J.A., Horvath C.M, & Malim M.H. (2017). Deep sequencing of HIV-1 reverse transcripts reveals the multifaceted anti-viral functions of APOBEC3G. Nature microbiology, 3(2), 220-233.

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Quantitative Viral Outgrowth Assay

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QVOA was performed as previously described [5 ,6 ]. Briefly, frozen PBMC were thawed and CD4+ cells were isolated using negative selecting bead purification (CD4+ T-cell Isolation Kit II, Miltenyi). The number of viable CD4+ T cells were counted and if the sample contained sufficient numbers rCD4 cells were isolated using negative selecting bead purification (anti-CD25, anti-biotin MicroBeads, anti-CD69 MicroBead Kit II, and anti–HLA-DR MicroBeads; Miltenyi). The resulting rCD4 cells were stimulated with Phytohaemagglutinin (PHA) and γ-irradiated PBMC and co-cultured with MOLT-4 cells transfected with CCR5 and naturally expressing CD4 and CXCR4 (MOLT4/CCR5; National Institutes of Health AIDS reagent program). The co-culture supernatants were tested for presence of HIV p24 by ELISA (PerkinElmer) after 14 or 21 days, to indicate outgrowth of replication-competent provirus [20 ]. For 2015 samples, all rCD4 cells were plated in a limiting dilution as previously described [20 ]. A portion of 2015 QVOA were tested for outgrowth viruses at 14 days, which was included and considered in both of the models. All time points collected after 2015, were measured at 21 days, with some also being tested at 14 days. In addition, QVOA for post-2015 samples were plated with a standard limited diluting plating strategy of approximately 14.5×10⁶ total rCD4 cells.

Ferreira R.C., Reynolds S.J., Capoferri A.A., Baker O., Brown E.E., Klock E., Miller J., Lai J., Saraf S., Kirby C., Lynch B., Hackman J., Gowanlock S.N., Tomusange S., Jamiru S., Anok A., Kityamuweesi T., Buule P., Bruno D., Martens C., Rose R., Lamers S.L., Galiwango R.M., Poon A.F., Quinn T.C., Prodger J.L, & Redd A.D. (2023). Temporary increase in circulating replication-competent latent HIV-infected resting CD4+ T cells after switch to an integrase inhibitor based antiretroviral regimen. medRxiv.

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Quantifying Viral Vector Titers

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Infective titers of lentiviral vectors (transducing units [TU]/mL) were determined using a quantitative polymerase chain reaction (qPCR)-based method.^{4 (link)} Lentiviral vector particle (vp) titer was analyzed by converting pg/mL results of p24 enzyme-linked immunosorbent assay (ELISA; PerkinElmer, Waltham, MA) to vp/mL by assuming 12,500 lentiviral particles per 1 pg of p24.^{12 ,13} Adenoviral vector particle titer was analyzed with high-performance liquid chromatography (HPLC).^{2 (link),14 (link)}

Leinonen H.M., Lepola S., Lipponen E.M., Heikura T., Koponen T., Parker N., Ylä-Herttuala S, & Lesch H.P. (2020). Benchmarking of Scale-X Bioreactor System in Lentiviral and Adenoviral Vector Production. Human Gene Therapy, 31(5-6), 376-384.

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HIV-1 Infection Assay in Activated PBMCs

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Peripheral blood mononuclear cells (PBMCs) were isolated from healthy donors’ buffy coats (Red Cross of Luxembourg, Luxembourg, Luxembourg) using Ficoll-Hypaque gradient as indicated previously (Sigma-Aldrich, Liège, Belgium). PBMCs were stimulated using 10 μg/mL phytohemagglutinin (PHA-P, Sigma Aldrich) for 48 h and recombinant IL-2 (10 U/mL, Roche, Sigma-Aldrich, Liège, Belgium) for another 24 h. Stimulated PBMCs were infected by the HIV-1 reference strains IIIB/ADA-M or primary clinical isolates expanded in culture from anonymized left-over samples (Centre Hospitalier de Luxembourg, Luxembourg, Luxembourg) in the presence or absence of drugs replaced every other day during 7 days. P24 production was measured in supernatants by ELISA (Perkin Elmer, Brussels, Belgium). Efavirenz (EFV) and azidothymidine (AZT) were obtained from Sigma_Aldrich. Enfuvirtide (T20) was purchased from Eurogentec (Seraing, Belgium).

Zheng Y., Yang X.W., Schols D., Mori M., Botta B., Chevigné A., Mulinge M., Steinmetz A., Schmit J.C, & Seguin-Devaux C. (2021). Active Components from Cassia abbreviata Prevent HIV-1 Entry by Distinct Mechanisms of Action. International Journal of Molecular Sciences, 22(9), 5052.

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