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Fluorescent Antibody Technique, Direct

The Fluorescent Antibody Technique, Direct is a powerful method used in biomedical research and diagnostics.
It involves the direct labeling of antibodies with fluorescent dyes, allowing for the visualization and localization of target antigens in cells or tissues.
This technique enables researchers to study the distribution and dynamics of specific proteins or molecules within complex biological samples.
By leveraging the sensitivity and specificity of fluorescent antibodies, scientists can gain valuable insights into cellular processes, disease mechanisms, and molecular interactions.
The Fluorescent Antibody Technique, Direct is a versitile tool that contributes to advancements in fields such as immunology, cell biology, and clinical diagnostics.

Most cited protocols related to «Fluorescent Antibody Technique, Direct»

1
Respiratory Virus Detection Using FilmArray
Cited 22 times
Viruses and bacteria used in this study are indicated in Table S1. Growth, quantification and verification of viral and bacterial cultures were performed by Zeptometrix (Buffalo, NY). Bocavirus (BoV) and Coronavirus (CoV) HKU1 could not be grown in culture. Instead, well-characterized clinical specimens were utilized and quantified in copies per ml by real-time PCR against a standard curve of synthetic template.
Residual clinical NPA specimens (stored frozen at −80°C) came from children younger than 18 years who had NPA collected for respiratory viral testing by direct fluorescent antibody (DFA) and culture at Primary Children's Medical Center (PCMC), Salt Lake City, UT between 2006 and 2008. Approximately half of the NPA specimens chosen for analysis were negative by DFA and viral culture. FilmArray testing was performed at both PCMC and ITI. PCR results were not used to inform clinical management or reported to microbiology technicians performing DFA and viral culture.
FilmArray data used for tuning the melt calling algorithm were acquired at sites performing beta testing of the instrument. The data used to validate the algorithm were acquired during clinical trials of the FilmArray system and RP pouch at the Medical University of South Carolina (Frederick S. Nolte, PhD), Detroit Medical Center (Hossein Salimnia, PhD), and Children's Medical Center of Dallas (Beverly Rogers, M.D.).
The institutional review boards of the University of Utah and PCMC approved this study and granted a waiver of informed consent because the patient samples were de-identified. All external clinical studies were performed with appropriate IRB approval. Data from these sites were de-identified before being sent to Idaho Technology.
Poritz M.A., Blaschke A.J., Byington C.L., Meyers L., Nilsson K., Jones D.E., Thatcher S.A., Robbins T., Lingenfelter B., Amiott E., Herbener A., Daly J., Dobrowolski S.F., Teng D.H, & Ririe K.M. (2011). FilmArray, an Automated Nested Multiplex PCR System for Multi-Pathogen Detection: Development and Application to Respiratory Tract Infection. PLoS ONE, 6(10), e26047.
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Publication 2011
Bacteria Bocavirus Buffaloes Child Coronavirus Infections Fluorescent Antibody Technique, Direct Freezing Patients Real-Time Polymerase Chain Reaction Respiratory Rate Sodium Chloride Virus Youth
2
Parainfluenza Virus Respiratory Infections
Cited 14 times
PIV detection was performed by conventional culture, direct fluorescent antibody tests, and/or reverse transcription polymerase chain reaction (RT-PCR) assay in respiratory samples. PIV upper respiratory tract infection (URTI) was defined as PIV detection in a nasopharyngeal or sputum sample, with URTI symptoms but no new pulmonary infiltrates. LRTD was divided into 3 groups: possible, probable, and proven. Possible LRTD was defined as PIV detection in a nasopharyngeal or sputum sample with new pulmonary infiltrates (but without confirmation of PIV in the lower respiratory tract) with or without LRTD signs or symptoms (eg, cough, wheezing, rales, tachypnea, shortness of breath, dyspnea, or hypoxia). Probable LRTD was defined as PIV detection in a bronchoalveolar lavage (BAL) or lung biopsy sample with LRTD symptoms, with or without pulmonary function decline, and without new pulmonary infiltrates. The definition of proven LRTD was PIV detection in a BAL or biopsy sample with new pulmonary infiltrates with or without LRTD symptoms.
Viral load was determined by quantitative RT-PCR using stored frozen repository samples [13 (link)]. Peak steroid dose was recorded from the period within 2 weeks before PIV infection in patients with URTI. In PIV LRTD cases, peak steroid doses were recorded from within 2 weeks before and after LRTD diagnosis, respectively, and exact steroid dose at 1 month after diagnosis was also collected. Death caused by respiratory failure was defined as any death caused exclusively or predominantly by respiratory failure [14 (link)].
Seo S., Xie H., Campbell A.P., Kuypers J.M., Leisenring W.M., Englund J.A, & Boeckh M. (2014). Parainfluenza Virus Lower Respiratory Tract Disease After Hematopoietic Cell Transplant: Viral Detection in the Lung Predicts Outcome. Clinical Infectious Diseases: An Official Publication of the Infectious Diseases Society of America, 58(10), 1357-1368.
Publication 2014
Biological Assay Biopsy Bronchoalveolar Lavage Cough Diagnosis Dyspnea Fluorescent Antibody Technique, Direct Freezing Hypoxia Infection Lung Nasopharynx Patients Respiratory Failure Respiratory Rate Respiratory System Reverse Transcriptase Polymerase Chain Reaction Sputum Steroids Upper Respiratory Infections
3
Autoantibody and Cytokine Profiling in Murine Lupus
Cited 10 times
HEp-2 immunofluorescence assays (Antibodies, Davis, CA) were performed as previously described (14 (link)) with serum diluted at 1/200 and were scored for relative fluorescence intensity of nuclear and cytoplasmic staining on a scale of 0–3 and for the presence or absence of mitotic chromatin by an observer blinded to the genotype of the mice.
For anti-nucleosome ELISA, polystyrene plates were coated with poly-L-lysine (Sigma-Aldrich, St. Louis, MO). Plates were then incubated with phenol-extracted and S1 nuclease-treated dsDNA from calf thymus (Sigma-Aldrich), followed by calf thymus histones type II-AS (Sigma-Aldrich). After blocking with 1% BSA in PBS, serial dilutions of serum from 1/200 to 1/5400 were added. Specific Abs were detected with alkaline phosphatase-conjugated goat anti-mouse IgG (Southern Bio-technology Associates, Birmingham, AL), and absorbance at 405/630 nm was compared with a purified PL2-3 monoclonal anti-nucleosome standard (27 (link)) for quantitation.
Anti-Sm ELISA was performed as previously described (14 (link)) with serial dilutions of serum from 1/200 to 1/5400. ELISAs for anti-ribosomal P Ag autoantibodies and anti-RNA Abs were described previously (28 (link)).
Anti-IgG2a RF titers were determined by ELISA essentially as described previously (29 (link)). Polystyrene plates were coated with purified mAb 23.3 (IgG2a,λ anti–4-hydroxy-3-nitro-phenylacetyl) overnight. After blocking with 1% BSA in PBS, serial dilutions of serum from 1/200 to 1/5400 were added. Specific Abs were detected with biotinylated anti-κ L chain (clone 187.1; BD Pharmingen, San Diego, CA), followed by alkaline-phosphatase–conjugated streptavidin (Invitrogen, Carlsbad, CA), and absorbance at 405/630 nm was compared with a 400tμ23 (IgM-RF) standard for quantitation.
Total serum IgG was determined by ELISA as described previously (14 (link)). Titers of individual IgG isotypes were measured by cytometric bead assay (Millipore, Bedford, MA) per the manufacturer’s protocols.
Serum IL-6, TNF-α, IL-12p40, IL-12p70, and IL-10 were measured by multiplex cytometric bead assay (Bio-Rad, Hercules, CA) per the manufacturer’s instructions. Serum IL-23(p19/p40) was measured by ELISA (eBioscience, San Diego, CA). Serum IFN-α was measured as previously described (14 (link)) with serum diluted 1/20.
Nickerson K.M., Christensen S.R., Shupe J., Kashgarian M., Kim D., Elkon K, & Shlomchik M.J. (2010). TLR9 Regulates TLR7- and MyD88-Dependent Autoantibody Production and Disease in a Murine Model of Lupus. Journal of immunology (Baltimore, Md. : 1950), 184(4), 1840-1848.
Publication 2010
Alkaline Phosphatase anti-IgG Anti-ribosomal P protein autoantibodies Antibodies Biological Assay Chromatin Clone Cells Cytoplasm DNA, Double-Stranded Enzyme-Linked Immunosorbent Assay Fluorescence Fluorescent Antibody Technique, Direct Genotype Goat Histones IgG2A IL10 protein, human Immunoglobulin Isotypes Interferon-alpha Interleukin-12 Interleukin-12 Subunit p40 Lysine Mus Nucleosomes Phenols Poly A Polystyrenes Ribosomes Serum Streptavidin Technique, Dilution Thymus Plant Tumor Necrosis Factor-alpha
4
Immunofluorescence Assay for Cleaved Caspase-3
Cited 9 times
For immunofluorescence assays, cells were cytospun onto positively charged slides (ThermoFisher Scientific, Waltham, MA), washed once with PBS, and fixed with 4% paraformaldehyde for 15 minutes. After washing three times with PBST, cells were permeabilized with 0.1% Triton/PBS, blocked for 15 minutes with 5% normal goat serum/PBST, and then incubated with the cleaved caspase 3 primary antibody (Cell Signaling Technology, Danvers, MA). Cells were then washed three times with PBST and incubated for 45 minutes in the dark with the Cy-3 conjugated secondary antibody (Jackson Immunoresearch, West Grove, PA). Following this incubation, cells were counterstained with DAPI and mounted with anti-fade mounting media (Vectastain, Burlingame, CA).
Weingart M.F., Roth J.J., Hutt-Cabezas M., Busse T.M., Kaur H., Price A., Maynard R., Rubens J., Taylor I., Mao X.G., Xu J., Kuwahara Y., Allen S.J., Erdreich-Epstein A., Weissman B.E., Orr B.A., Eberhart C.G., Biegel J.A, & Raabe E.H. (2014). Disrupting LIN28 in atypical teratoid rhabdoid tumors reveals the importance of the mitogen activated protein kinase pathway as a therapeutic target. Oncotarget, 6(5), 3165-3177.
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Publication 2014
Caspase 3 Cells DAPI Fluorescent Antibody Technique, Direct Goat Immunoglobulins paraform Serum
5
Genetic Manipulation and Analysis of C. elegans
Cited 8 times

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Publication 2014
Alleles Animals Biological Assay Caenorhabditis elegans Cloning Vectors Co-Immunoprecipitation Deletion Mutation Escherichia coli Fluorescent Antibody Technique, Direct Genes Gene Therapy, Somatic Genome Germ Line RNA, Double-Stranded RNA Interference Saccharomyces cerevisiae Strains Transgenes Western Blot Yeast Two-Hybrid System Techniques

Most recents protocols related to «Fluorescent Antibody Technique, Direct»

1
Quantifying Notch1 Expression in Cancer Spheres
Immunofluorescence assays were performed to determine the Notch1 expression of the vehicle and ASR490-treated first, second, and third-generation spheres as per the protocol described earlier (Suman et al., 2014 (link)). The spheres were imaged using a KEYENCE fluorescence microscope (BZ-X800/BZ-X810).
Saran U., Chandrasekaran B., Tyagi A., Shukla V., Singh A., Sharma A.K, & Damodaran C. (2023). A small molecule inhibitor of Notch1 modulates stemness and suppresses breast cancer cell growth. Frontiers in Pharmacology, 14, 1150774.
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Publication 2023
Fluorescent Antibody Technique, Direct Microscopy, Fluorescence
2
Immunofluorescence Imaging of ER-Localized Proteins
Immunofluorescence assays were performed as described in the previous study.42 (link)Cy3-labeled DICAR, FITC-labeled TnnT were used as described by the manufacturer’s instructions.42 (link) FL glibenclamide was used to mark the ER. The live cells were stained by FL Glibenclamide and fixed by the DICAR FISH probe A. The FV10i Confocal Microscope (Olympus, Japan) was used to capture the images.
Yuan Q., Sun Y., Yang F., Yan D., Shen M., Jin Z., Zhan L., Liu G., Yang L., Zhou Q., Yu Z., Zhou X., Yu Y., Xu Y., Wu Q., Luo J., Hu X, & Zhang C. (2023). CircRNA DICAR as a novel endogenous regulator for diabetic cardiomyopathy and diabetic pyroptosis of cardiomyocytes. Signal Transduction and Targeted Therapy, 8, 99.
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Publication 2023
Cells Fishes Fluorescein-5-isothiocyanate Fluorescent Antibody Technique, Direct Glyburide Microscopy, Confocal Troponin T
3
Immunofluorescence analysis of cell signaling
In this study, we used 24-well sterile slides to culture MRC-5 cells for 24 h. After allowing the cells to fuse to 60–70%, IL-27 (100 ng/mL) and/or TGF-β1 (40 ng/mL) were used to treat the cells for 48 h. Immunofluorescence assays were performed according to previous studies [29 (link)]. When incubation was complete, cells were washed three times with prechilled PBS before fixation with immunostaining fixative (Beyotime) for 30 min. Subsequently, cells were incubated with Triton X-100 (Beyotime) permeabilization buffer for 15 min and then blocked with QuickBlock™ (Beyotime) for 30 min at 37 °C. Then, primary antibodies against α-SMA, FN, COL1, LC3B and Beclin1 were incubated with the cells at 4 °C for 12 h. Subsequently, the appropriate fluorescein-conjugated secondary antibody was added to the cells and incubated. The cell nuclei were stained with DAPI. The samples were observed with a confocal microscope and photographed for analysis.
Ting L., Feng Y., Zhou Y., Tong Z, & Dong Z. (2023). IL-27 induces autophagy through regulation of the DNMT1/lncRNA MEG3/ERK/p38 axis to reduce pulmonary fibrosis. Respiratory Research, 24, 67.
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Publication 2023
Antibodies BECN1 protein, human Buffers Cell Culture Techniques Cell Nucleus Cells DAPI Fixatives Fluorescein Fluorescent Antibody Technique, Direct Immunoglobulins Interleukin-27 Microscopy, Confocal Sterility, Reproductive TGF-beta1 Triton X-100
4
Monitoring Transplant Patients' Health
Laboratory tests, including complete blood count (CBC) and liver function test, were periodically performed after LT, initially 4 times a day. The sampling interval was gradually extended to once-daily during admission. After discharge, patients were checked monthly at routine outpatient department visits.
For infection surveillance, quantitative CMV antigenemia targeting pp65 was tested using direct immunofluorescence method (CINA Kit system, Argene Biosoft, Varilhes, France) 3 times during the first week after transplantation and then weekly during hospitalization. After discharge, patients were monitored monthly during the first year after LT. CMV qPCR testing (Real-Q assay; BioSewoom, Seoul, South Korea) was performed when the patient had leukopenia, according to the clinician’s judgement.
Ha C., Kim S.J., Kim J.M., Joh J.W., Jang K.T., Choi G.S, & Kang E.S. (2023). Detecting Donor-Derived DNA by Real-Time PCR in Recipients Suspected of Graft-Versus-Host-Diseases After Liver Transplantation: A Case Series and Literature Review. Annals of Transplantation, 28, e938287-1-e938287-11.
Publication 2023
Biological Assay Fluorescent Antibody Technique, Direct Hospitalization Infection Leukopenia Liver Function Tests Outpatients Patient Discharge Patients Transplantation UL83 protein, Human herpesvirus 5
5
SV40 DNA Transfection and Lytic Induction
iD98/HR1 cells were plated on coverslips in 6-well plates and treated with 100 μg/mL of PAA, 24 h prior to being mock transfected or transfected with isolated SV40 DNA. SV40 DNA isolation was carried out as previously described (64 (link)). The transfection mix was incubated with the cells for 4 h in the 37°C cell incubator (humidified, with 5% CO2), and then the cells were washed with 2 mL of DMEM with 10% FBS (D10F) before another 2 mL of D10F was added as growth medium. The cells were grown for 1 h, and then 100 μg/mL PAA was added back to the medium. Cells were grown for another 2 h and then were induced to enter EBV’s lytic phase by the addition of 200 nM 4-OHT. At 48 h after transfection, the cells were incubated with 10 μM EdU for 1 h and then fixed for click chemistry and immunofluorescence assays.
Rosemarie Q., Kirschstein E, & Sugden B. (2023). How Epstein-Barr Virus Induces the Reorganization of Cellular Chromatin. mBio, 14(1), e02686-22.
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Publication 2023
Cells Fluorescent Antibody Technique, Direct isolation Simian virus 40 Transfection

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DAPI is a fluorescent dye that binds strongly to adenine-thymine (A-T) rich regions in DNA. It is commonly used as a nuclear counterstain in fluorescence microscopy to visualize and locate cell nuclei.
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More about "Fluorescent Antibody Technique, Direct"

The Fluorescent Antibody Technique, Direct is a powerful and versatile method used in biomedical research and clinical diagnostics.
Also known as immunofluorescence or direct immunofluorescence, this technique involves the direct labeling of antibodies with fluorescent dyes, enabling the visualization and localization of target antigens within cells or tissues.
By leveraging the specificity and sensitivity of fluorescent antibodies, scientists can gain valuable insights into cellular processes, disease mechanisms, and molecular interactions.
The Fluorescent Antibody Technique, Direct is widely used in various fields, including immunology, cell biology, and clinical diagnostics.
It allows researchers to study the distribution and dynamics of specific proteins or molecules within complex biological samples, such as cell cultures or tissue sections.
This technique is often combined with other fluorescent probes, such as DAPI (4',6-diamidino-2-phenylindole) for nuclear staining, Alexa Fluor 488 for green fluorescence, and various blocking agents like FBS (Fetal Bovine Serum) and Triton X-100 to minimize non-specific binding.
The process typically involves the fixation and permeabilization of cells or tissues, followed by the incubation with fluorescently-labeled primary antibodies that bind to the target antigens.
The samples are then visualized using advanced microscopy techniques, such as confocal laser scanning microscopy (e.g., LSM 700), which enables the acquisition of high-resolution, three-dimensional images.
The fluorescent signals can be further enhanced using mounting media like ProLong Gold antifade reagent.
The Fluorescent Antibody Technique, Direct is a versatile tool that has contributed to advancements in various fields, including the study of protein localization, cell signaling pathways, and disease biomarkers.
It is also widely used in diagnostic applications, such as the detection of infectious agents, tumor markers, and other clinically relevant targets.
Researchers can further optimize their experimental protocols by leveraging resources like PubCompare.ai, which uses AI-powered analysis to help identify the best protocols from literature, preprints, and patents.