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> Chemicals & Drugs > Amino Acid > Streptavidin

Streptavidin

Streptavidin is a tetrameric protein derived from the bacterium Streptomyces avidinii.
It exhibits a high affinity for the vitamin biotin, forming a strong non-covalent bond.
This property makes streptavidin a versatile tool in biotechnology and biomedical research, enabling the development of sensitive and specific assays, purification techniques, and labeling methods.
Streptavidin is commonly used to detect and localize biotinylated molecules, such as proteins, nucleic acids, and lipids, in a wide range of applications, including immunoassays, affinity chromatography, and floresscence microscopy.
The robust and stable nature of the streptavidin-biotin interaction, as well as the commercial availability of a variety of streptavidin-conjugated reagents, have made streptavidin an indispensable component of many experimental protocols.

Most cited protocols related to «Streptavidin»

1
Immunostaining for viral antigen detection
The streptavidin alkaline phosphatase method was adapted to detect the viral antigen using a polyclonal anti-ZIKV antibody produced at the Evandro Chagas Institute2 (link). The biotin-streptavidin peroxidase method was used for immunostaining of tissues with antibodies specific for each marker studied. First, the tissue samples were deparaffinized in xylene and hydrated in a decreasing ethanol series (90%, 80%, and 70%). Endogenous peroxidase was blocked by incubating the sections in 3% hydrogen peroxide for 45 min. Antigen retrieval was performed by incubation in citrate buffer, pH 6.0, or EDTA, pH 9.0, for 20 min at 90 °C. Nonspecific proteins were blocked by incubating the sections in 10% skim milk for 30 min. The histological sections were then incubated overnight with the primary antibodies diluted in 1% bovine serum albumin (Supplementary Table S1). After this period, the slides were immersed in 1 × PBS and incubated with the secondary biotinylated antibody (LSAB, DakoCytomation) in an oven for 30 min at 37 °C. The slides were again immersed in 1X PBS and incubated with streptavidin peroxidase (LSAB, DakoCytomation) for 30 min at 37 °C. The reactions were developed with 0.03% diaminobenzidine and 3% hydrogen peroxide as the chromogen solution. After this step, the slides were washed in distilled water and counterstained with Harris hematoxylin for 1 min. Finally, the sections were dehydrated in an increasing ethanol series and cleared in xylene.
Azevedo R.S., de Sousa J.R., Araujo M.T., Martins Filho A.J., de Alcantara B.N., Araujo F.M., Queiroz M.G., Cruz A.C., Vasconcelos B.H., Chiang J.O., Martins L.C., Casseb L.M., da Silva E.V., Carvalho V.L., Vasconcelos B.C., Rodrigues S.G., Oliveira C.S., Quaresma J.A, & Vasconcelos P.F. (2018). In situ immune response and mechanisms of cell damage in central nervous system of fatal cases microcephaly by Zika virus. Scientific Reports, 8, 1.
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Publication 2018
Alkaline Phosphatase Antibodies Antibodies, Anti-Idiotypic Antigens Antigens, Viral azo rubin S Biotin Buffers Citrates Edetic Acid Ethanol Hematoxylin Immunoglobulins Milk, Cow's Peroxidase Peroxide, Hydrogen Peroxides Proteins Serum Albumin, Bovine Streptavidin Tissues Tritium Xylene Zika Virus
2
High-Throughput Chromatin Conformation Capture

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Publication 2014
Cell Lines Cell Nucleus Cells Formaldehyde Ligation Microtubule-Associated Proteins Nucleotides Streptavidin Technique, Dilution
3
Single-Strand DNA Library for Ancient Genomics
DNA libraries for sequencing are normally prepared from double-stranded DNA (Fig. 1). However, for ancient DNA the use of single-stranded DNA may be advantageous as it will double its representation in the library. Furthermore, in a single-stranded DNA library, double-stranded molecules that carry modifications on one strand that prevent their incorporation into double-stranded DNA libraries could still be represented by the unmodified strand. We therefore devised a single-stranded library preparation method wherein the ancient DNA is dephosphorylated, heat denatured, and ligated to a biotinylated adaptor oligonucleotide, which allows its immobilization on streptavidin-coated beads (Fig. 1). A primer hybridized to the adaptor is then used to copy the original strand with a DNA polymerase. Finally, a second adaptor is joined to the copied strand by blunt-end ligation and the library molecules are released from the beads. The entire protocol is devoid of DNA purification steps, which inevitably cause loss of material.
We applied this method to aliquots of the two DNA extracts (as well as side fractions) that were previously generated from the 40 mg of bone that comprised the entire inner part of the phalanx (2 (link), 8 ). Comparisons of these newly generated libraries to the two libraries generated in the previous study (2 (link)) show at least a 6-fold and 22-fold increase in the recovery of library molecules (8 ), which is particularly pronounced for longer molecules (Fig. S4).
In addition to improved sequence yield, the single-strand library protocol reveals new aspects of DNA fragmentation and modification patterns (8 ). Since the ends of both DNA strands are left intact, it reveals that strand breakage occurs preferentially before and after guanine residues (Fig. S6), suggesting that guanine nucleotides are frequently lost from ancient DNA, possibly as the result of depurination. It also reveals that deamination of cytosine residues occurs with almost equal frequencies at both ends of the ancient DNA molecules. Since deamination is hypothesized to be frequent in single-stranded DNA overhangs (9 (link), 10 (link)), this suggests that 5′- and 3′-overhangs occur at similar lengths and frequencies in ancient DNA.
Meyer M., Kircher M., Gansauge M.T., Li H., Racimo F., Mallick S., Schraiber J.G., Jay F., Prüfer K., de Filippo C., Sudmant P.H., Alkan C., Fu Q., Do R., Rohland N., Tandon A., Siebauer M., Green R.E., Bryc K., Briggs A.W., Stenzel U., Dabney J., Shendure J., Kitzman J., Hammer M.F., Shunkov M.V., Derevianko A.P., Patterson N., Andrés A.M., Eichler E.E., Slatkin M., Reich D., Kelso J, & Pääbo S. (2012). A HIGH COVERAGE GENOME SEQUENCE FROM AN ARCHAIC DENISOVAN INDIVIDUAL. Science (New York, N.Y.), 338(6104), 222-226.
Publication 2012
Bones Bones of Fingers Cytosine Deamination DNA DNA, Ancient DNA, Double-Stranded DNA, Single-Stranded DNA-Directed DNA Polymerase DNA Fragmentation DNA Library Guanine Guanine Nucleotides Immobilization Ligation Oligonucleotide Primers Oligonucleotides Streptavidin
4
Panda Genome Assembly and Annotation
We constructed the paired-end DNA libraries with insert sizes larger than 2 kb by self-ligation of the DNA fragments and merging the two ends of the DNA fragment. We randomly fragmented the circularized DNA and enriched the fragments crossing the merged boundaries using magnetic beads with biotin and streptavidin. The sequencing process followed the manufacturer’s instructions (Illumina), and the fluorescent images were processed to sequences using the Illumina data processing pipeline (v1.1).
The genome sequence was assembled with short reads using SOAPdenovo software6 (http://soap.genomics.org.cn), which adopts the de Bruijn graph data structure to construct contigs7 (link). The reads were then realigned to the contig sequence, and the paired-end relationship between the reads was transferred to linkage between contigs. We constructed scaffolds starting with short paired-ends and then iterated the scaffolding process, step by step, using longer insert size paired-ends. To fill the intra-scaffold gaps, we used the paired-end information to retrieve read pairs that had one read well-aligned on the contigs and another read located in the gap region, then did a local assembly for the collected reads.
Known transposable elements were identified using RepeatMasker (version 3.2.6)14 against the Repbase31 (link) transposable element library (version 2008-08-01), and highly diverged transposable elements were identified with RepeatProteinMask14 by aligning the genome sequence to the curated transposable-element-related proteins. A de novo panda repeat library was constructed using RepeatModeller14 . Using evidence-based gene prediction, the human and dog genes (Ensembl release 52) were projected onto the panda genome, and the gene loci were defined by using both sequence similarity and whole-genome synteny information. De novo gene prediction was performed using Genscan16 (link) and Augustus17 (link). A reference gene set was created by merging all of the gene sets. The sequencing reads were mapped on the panda genome sequence using SOAPaligner8 (link), and heterozygous SNPs were identified by SOAPsnp9 (link).
Li R., Fan W., Tian G., Zhu H., He L., Cai J., Huang Q., Cai Q., Li B., Bai Y., Zhang Z., Zhang Y., Wang W., Li J., Wei F., Li H., Jian M., Li J., Zhang Z., Nielsen R., Li D., Gu W., Yang Z., Xuan Z., Ryder O.A., Leung F.C., Zhou Y., Cao J., Sun X., Fu Y., Fang X., Guo X., Wang B., Hou R., Shen F., Mu B., Ni P., Lin R., Qian W., Wang G., Yu C., Nie W., Wang J., Wu Z., Liang H., Min J., Wu Q., Cheng S., Ruan J., Wang M., Shi Z., Wen M., Liu B., Ren X., Zheng H., Dong D., Cook K., Shan G., Zhang H., Kosiol C., Xie X., Lu Z., Zheng H., Li Y., Steiner C.C., Lam T.T., Lin S., Zhang Q., Li G., Tian J., Gong T., Liu H., Zhang D., Fang L., Ye C., Zhang J., Hu W., Xu A., Ren Y., Zhang G., Bruford M.W., Li Q., Ma L., Guo Y., An N., Hu Y., Zheng Y., Shi Y., Li Z., Liu Q., Chen Y., Zhao J., Qu N., Zhao S., Tian F., Wang X., Wang H., Xu L., Liu X., Vinar T., Wang Y., Lam T.W., Yiu S.M., Liu S., Zhang H., Li D., Huang Y., Wang X., Yang G., Jiang Z., Wang J., Qin N., Li L., Li J., Bolund L., Kristiansen K., Wong G.K., Olson M., Zhang X., Li S., Yang H., Wang J, & Wang J. (2009). The sequence and de novo assembly of the giant panda genome. Nature, 463(7279), 311-317.
Publication 2009
Amino Acid Sequence Biotin DNA Library DNA Transposable Elements Genes Genetic Loci Genome Heterozygote Homo sapiens Ligation Selfish DNA Single Nucleotide Polymorphism Streptavidin Synteny
5
Retinal Vasculature Staining Protocol
Postnatal P6 eyes were briefly fixed with 2% PFA in PBS at 4°C, then retinas were dissected and stored in methanol at -20°C. Immediately prior to staining retinas were re-fixed in 4% PFA for 20 minutes. PBS-washed retinas were stained with biotinylated isolectin B4 (Vectorlabs) followed by streptavidin-alexafluor 568 (Molecular Probes) and flat mounted for epifluorescence analysis on a CellR microscope (Olympus).
Zudaire E., Gambardella L., Kurcz C, & Vermeren S. (2011). A Computational Tool for Quantitative Analysis of Vascular Networks. PLoS ONE, 6(11), e27385.
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Publication 2011
Eye Isolectins Methanol Microscopy Molecular Probes Retina Streptavidin

Most recents protocols related to «Streptavidin»

1
Translocation of Surface Lipoproteins in E. coli
Not available on PMC !

Example 6

TbpB and NMB0313 genes were amplified from the genome of Neisseria meningitidis serotype B strain B16B6. The LbpB gene was amplified from Neisseria meningitidis serotype B strain MC58. Full length TbpB was inserted into Multiple Cloning Site 2 of pETDuet using restriction free cloning ((F van den Ent, J. Löwe, Journal of Biochemical and Biophysical Methods (Jan. 1, 2006)).). NMB0313 was inserted into pET26, where the native signal peptide was replaced by that of pelB. Mutations and truncations were performed on these vectors using site directed mutagenesis and restriction free cloning, respectively. Pairs of vectors were transformed into E. coli C43 and were grown overnight in LB agar plates supplemented with kanamycin (50 μg/mL) and ampicillin (100 μg/mL).

tbpB genes were amplified from the genomes of M. catarrhalis strain 035E and H. influenzae strain 86-028NP and cloned into the pET52b plasmid by restriction free cloning as above. The corresponding SLAMs (M. catarrhalis SLAM 1, H. influenzae SLAM1) were inserted into pET26b also using restriction free cloning. A 6His-tag was inserted between the pelB and the mature SLAM sequences as above. Vectors were transformed into E. coli C43 as above.

Cells were harvested by centrifugation at 4000 g and were twice washed with 1 mL PBS to remove any remaining growth media. Cells were then incubated with either 0.05-0.1 mg/mL biotinylated human transferrin (Sigma-aldrich T3915-5 MG), α-TbpB (1:200 dilution from rabbit serum for M. catarrhalis and H. influenzae; 1:10000 dilution from rabbit serum for N. meningitidis), or α-LbpB (1:10000 dilution from rabbit serum-obtained a gift from J. Lemieux) or α-fHbp (1:5000 dilution from mouse, a gift from D. Granoff) for 1.5 hours at 4° C., followed by two washes with 1 mL of PBS. The cells were then incubated with R-Phycoerythrin-conjugated Streptavidin (0.5 mg/ml Cedarlane) or R-phycoerythrin conjugated Anti-rabbit IgG (Stock 0.5 mg/ml Rockland) at 25 ug/mL for 1.5 hours at 4° C. The cells were then washed with 1 mL PBS and resuspended in 200 uL fixing solution (PBS+2% formaldehyde) and left for 20 minutes. Finally, cells were washed with 2×1 mL PBS and transferred to 5 mL polystyrene FACS tubes. The PE fluorescence of each sample was measured for PE fluorescence using a Becton Dickinson FACSCalibur. The results were analyzed using FLOWJO software and were presented as mean fluorescence intensity (MFI) for each sample. For N. meningtidis experiments, all samples were compared to wildtype strains by normalizing wildtype fluorescent signals to 100%. Errors bars represent the standard error of the mean (SEM) across three experiments. Results were plotted statistically analysed using GraphPad Prism 5 software. The results shown in FIG. 6 for the SLPs, TbpB (FIG. 6A), LbpB. (FIG. 6B) and fHbp (FIG. 6C) demonstrate that SLAM effects translocation of all three SLP polypeptides in E. coli. The results shown in FIG. 10 demonstrate that translocation of TbpB from M. catarrhalis (FIG. 10C) and in H. influenzae (FIG. 10D) in E. coli require the co-expression of the required SLAM protein (Slam is an outer membrane protein that is required for the surface display of lipidated virulence factors in Neisseria. Hooda Y, Lai C C, Judd A, Buckwalter C M, Shin H E, Gray-Owen S D, Moraes T F. Nat Microbiol. 2016 Feb. 29; 1:16009).

US11872275B2. SLAM polynucleotides and polypeptides and uses thereof (2024-01-16). ENGINEERED ANTIGENS INC. [CA]. Inventors: Trevor F. Moraes [CA], Christine Chieh-Lin Lai [CA], Yogesh Hooda [CA], Andrew Judd [CA], Anthony B. Schryvers [CA], Scott D. Gray-Owen [CA].
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Patent 2024
ADRB2 protein, human Agar Ampicillin anti-IgG Cells Centrifugation Cloning Vectors Culture Media Escherichia coli Fluorescence Formaldehyde Genes Genome Haemophilus influenzae Homo sapiens Kanamycin Lipoproteins Membrane Proteins Moraxella catarrhalis Mus Mutagenesis, Site-Directed Mutation Neisseria Neisseria meningitidis Phycoerythrin Plasmids Polypeptides Polystyrenes prisma Rabbits Serum Signaling Lymphocytic Activation Molecule Family Member 1 Signal Peptides Strains Streptavidin Technique, Dilution Transferrin Translocation, Chromosomal Virulence Factors
2
Simultaneous CD40 and CEACAM5 Targeting
Not available on PMC !

Example 4

Aim

The aim of the study was to evaluate the ability of selected CD40 and CEACAM5 targeting RUBY™ bsAbs to bind both their targets simultaneously as well as their potential cross-reactivity with additional members of the CEA protein family was evaluated by ELISA.

Materials and Methods

96-well plates were coated with 0.5 μg/mL antigen, hCEACAM-1 (2244-CM-050, R&D Systems), hCEACAM-5 (4128-CM-050, R&D Systems), hCEACAM-6 (3934-CM-050, R&D Systems) or CEACAM-8 (9639-CM-050, R&D Systems) in PBS over night at 4° C. After washing in PBS/0.05% Tween 20 (PBST), the plates were blocked with PBST, 2% BSA for at least 30 minutes at room temperature before a second round of washing. RUBY bsAbs, diluted in PBST, 0.5% BSA, were then added and allowed to bind for at least 1 hour at room temperature. After washing, plates were incubated with either 50 μl detection antibody (0.5 μg/ml HRP conjugated goat anti human-kappa light chain, #STAR127P, AbD Serotec) for analysis of binding to CEACAM protein family proteins or 0.5 μg/ml biotinylated hCD40-muIg (504-030, Ancell) followed by HRP conjugated streptavidin (21126, Pierce) for confirmation of dual antigen binding. Finally, a final round of washing was performed and bound complexes detected using SuperSignal Pico Luminescent as substrate and luminescence signals were measured using Fluostar Optima.

Results and Conclusions

All evaluated RUBY™ bsAbs was indeed able to bind to both CD40 and human CEACAM5 simultaneously (FIG. 2), although with varying potency. In general, bsAbs carrying 1132 as CD40 binding antibody (Multi46-Multi49) displayed lower potency in the dual target ELISA, as compared to bsAbs carrying G12_mut. Also, Multi38 displayed reduced dual target binding compared to other G12_mut based bsAbs, likely due to lower CEACAM5 binding of Fab6 than other evaluated CEACAM5 binding antibodies.

As can be seen in FIG. 3, a majority of the evaluated CD40 and CEACAM5 targeting RUBY™ bsAbs did not cross react with any of the other CEA family members evaluated. However, a limited number of the assayed bsAb did show significant cross-reactivity with CEACAM1 (Multi38, Multi39, Multi45 and Multi 49) or CEACAM6 (Multi40).

All in all, it can be concluded that all evaluated RUBY™ bsAbs have the ability to bind CD40 and CEACAM5 simultaneously and a majority of the set was specific for CEACAM5, with no or little detectable binding to other evaluated members of the CEA protein family.

US11873348B2. Peptides (2024-01-16). ALLIGATOR BIOSCIENCE AB [SE]. Inventors: Peter Ellmark [SE], Karin Hagerbrand [SE], Laura Varas [SE], Mattias Levin [SE], Anna Säll [SE], Laura von Schantz [SE].
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Patent 2024
Antibodies Antigens biliary glycoprotein I Binding Proteins Carcinoembryonic Antigen carcinoembryonic antigen-related cell adhesion molecule 6, human Cross Reactions Enzyme-Linked Immunosorbent Assay Family Member Gene Products, Protein Goat Homo sapiens Immunoglobulin kappa-Chains Immunoglobulins Luminescence Streptavidin Tween 20 Vision
3
PHESELECTOR Assay for Thiol Reactivity Evaluation
Not available on PMC !

Example 2

Bovine serum albumin (BSA), erbB2 extracellular domain (HER2) and streptavidin (100 μl of 2 μg/ml) were separately coated on Maxisorp 96 well plates. After blocking with 0.5% Tween-20 (in PBS), biotinylated and non-biotinylated hu4D5Fabv8-ThioFab-Phage (2×1010 phage particles) were incubated for 1 hour at room temperature followed by incubation with horseradish peroxidase (HRP) labeled secondary antibody (anti-M13 phage coat protein, pVIII protein antibody). FIG. 8 illustrates the PHESELECTOR Assay by a schematic representation depicting the binding of Fab or ThioFab to HER2 (top) and biotinylated ThioFab to streptavidin (bottom).

Standard HRP reaction was carried out and the absorbance was measured at 450 nm. Thiol reactivity was measured by calculating the ratio between OD450 for streptavidin/OD450 for HER2. A thiol reactivity value of 1 indicates complete biotinylation of the cysteine thiol. In the case of Fab protein binding measurements, hu4D5Fabv8 (2-20 ng) was used followed by incubation with HRP labeled goat polyclonal anti-Fab antibodies.

US11873330B2. Cysteine engineered antibodies and conjugates (2024-01-16). Genentech, Inc. [None]. Inventors: Sunil Bhakta [US], Jagath R. Junutula [US].
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Patent 2024
Anti-Antibodies Bacteriophage M13 Bacteriophages Biological Assay Biotinylation Cardiac Arrest Cysteine ERBB2 protein, human Goat herstatin protein, human Horseradish Peroxidase Immunoglobulins Proteins Serum Albumin, Bovine Streptavidin Sulfhydryl Compounds Tween 20
4
Peptide Synthesis Reagents and Cell Culture Protocols
Not available on PMC !

Example 1

Reagents for peptide synthesis were purchased from Chem-Impex (Wood Dale, IL), NovaBiochem (La Jolla, CA), or Anaspec (San Jose, CA). Rink amide resin LS (100-200 mesh, 0.2 mmol/g) was purchased from Advanced ChemTech. Cell culture media, fetal bovine serum, penicillin-streptomycin, 0.25% trypsin-EDTA, and DPBS were purchased from Invitrogen (Carlsbad, CA). Methyl 3,5-dimethylbenzoiate, N-bromosuccinimide, diethyl phosphite, 2,2′-dipyridyl disulfide, and other organic reagents/solvents were purchased from Sigma-Aldrich (St. Louis, MO). Anti-GST-Tb and streptavidin-d2 were purchased from Cisbio (Bedford, MA). The NF-κB reporter (Luc)-HEK293 cell line and One-Step™ luciferase assay system were purchased from BPS Bioscience (San Diego, CA).

[Figure (not displayed)]

US11878046B2. Di-sulfide containing cell penetrating peptides and methods of making and using thereof (2024-01-23). Ohio State Innovation Foundation [US]. Inventors: Dehua Pei [US], Ziqing Qian [US].
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Patent 2024
Anabolism Biological Assay Bromosuccinimide Cell Culture Techniques Cells Culture Media Disulfides Edetic Acid Fetal Bovine Serum HEK293 Cells Luciferases Penicillins Peptide Biosynthesis Phosphite RELA protein, human Rink amide resin Solvents Streptavidin Streptomycin Trypsin
5
Immunofluorescence Analysis of Muscle Tissue
The muscles were cut on a cryostat at − 23 °C (7 μm), air-dried, and stored at − 20 °C. Slides were air-dried, rehydrated, and fixed in 4% paraformaldehyde (PFA) for 20 min at the time of staining. For CD63/DAPI/laminin staining, sections were incubated with mouse anti-CD63 IgG1 antibody (1:100 dilution, ab108950, Abcam, Cambridge, UK) and rabbit anti-laminin IgG antibody (1:100 dilution, L9393, Sigma-Aldrich, St. Louis, MO) overnight at 4 °C. Slides were washed in PBS, then incubated with Alexa Fluor 488 goat anti-mouse IgG1 (1:250 dilution, A11001, Invitrogen, Waltham, MA) and Alexa Fluor 594 goat anti-rabbit IgG (1:250 dilution, A11012, Invitrogen) secondary antibodies for 1 h at room temperature. Slides were washed in PBS and mounted with VectaShield fluorescent mounting media with DAPI (H-1200-10, Vector Laboratories, Newark, CA). For CD9/DAPI/dystrophin staining, sections were incubated with rabbit anti-CD9 IgG (1:100 dilution, SA35-08, Invitrogen) and mouse anti-dystrophin IgG2b (1:250 dilution, 08168, Sigma-Aldrich) overnight, followed by incubation with Alexa Fluor 594 goat anti-rabbit IgG (1:250 dilution, A11012, Invitrogen) and Alexa Fluor 647 goat anti-mouse IgG2b (1:250 dilution, A32728, Invitrogen) for 1 h at room temperature. For CD81/DAPI/dystrophin staining, sections were incubated with rabbit anti-CD81(1:100 dilution, SN206-01, Novus Biologicals, Centennial, CO) and mouse anti-dystrophin IgG2b (1:250 dilution, 08168, Sigma-Aldrich) overnight, followed by incubation with Alexa Fluor 594 goat anti-rabbit IgG (1:250 dilution, A11012, Invitrogen) and Alexa Fluor 647 goat anti-mouse IgG2b (1:250 dilution, A32728, Invitrogen) for 1 h at room temperature. For Pax7/CD9/DAPI/WGA staining, sections were subjected to epitope retrieval using sodium citrate (10 mM, pH 6.5) at 92 °C, followed by blocking of endogenous peroxidase activity with 3% hydrogen peroxide in PBS. Sections were incubated overnight in mouse anti-Pax7 IgG1 (1:100 dilution, Developmental Studies Hybridoma Bank, Iowa City, IA) and rabbit anti-CD9 IgG (1:100 dilution, SA35-08, Invitrogen), followed by incubation in goat anti-mouse biotin-conjugated secondary antibody (dilution 1:1,000, 115-065-205; Jackson ImmunoResearch, West Grove, PA) and Alexa Fluor 647 goat anti-rabbit IgG (1:250 dilution, A32733, Invitrogen) for 1 h at room temperature. Next, sections were incubated with streptavidin-HRP (1:500 dilution, S-911, Invitrogen) and Texas Red-conjugated Wheat Germ Agglutinin (WGA) (1:50 dilution, W21405, Invitrogen) at room temperature for 1 h, before incubation in Tyramide Signal Amplification (TSA) Alexa Fluor 488 (1:500 dilution, B40953, Invitrogen). Sections were mounted with VectaShield fluorescent mounting media with DAPI (H-1200-10, Vector Laboratories).
Images were captured with a Zeiss upright microscope (AxioImager M1, Oberkochen, Germany). To quantify the percentage of nuclei (DAPI+) expressing CD63, MyoVision software was used for automated analysis of nuclear density in cross-sections [39 (link)], and nuclei-expressing CD63 (identified as DAPI+/CD63+ events) were counted manually in a blinded manner by the same assessor for all sections using the Zen Blue software.
Ismaeel A., Van Pelt D.W., Hettinger Z.R., Fu X., Richards C.I., Butterfield T.A., Petrocelli J.J., Vechetti I.J., Confides A.L., Drummond M.J, & Dupont-Versteegden E.E. (2023). Extracellular vesicle distribution and localization in skeletal muscle at rest and following disuse atrophy. Skeletal Muscle, 13, 6.
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Publication 2023
Alexa594 alexa fluor 488 Alexa Fluor 647 anti-IgG Antibodies Antibodies, Anti-Idiotypic Biological Factors Biotin Cardiac Arrest Cell Nucleus Cloning Vectors DAPI DMD protein, human Epitopes Goat Hybridomas IgG1 IgG2B Immunoglobulins Laminin Microscopy Mus Muscle Tissue Novus paraform PAX7 protein, human Peroxidase Peroxides Rabbits Sodium Citrate Streptavidin Technique, Dilution Tritium Wheat Germ Agglutinins

Top products related to «Streptavidin»

1
Dynabeads m 280 streptavidin by Thermo Fisher Scientific
Sourced in United States, Norway, China, France, Germany, Japan
Dynabeads M-280 Streptavidin are uniform superparamagnetic beads coated with streptavidin, a protein that binds strongly to biotin. They are designed for efficient isolation and purification of biotinylated molecules, such as nucleic acids, proteins, and cells.
2
Bovine serum albumin (bsa) by Merck Group
Sourced in United States, Germany, United Kingdom, China, Italy, Japan, France, Sao Tome and Principe, Canada, Macao, Spain, Switzerland, Australia, India, Israel, Belgium, Poland, Sweden, Denmark, Ireland, Hungary, Netherlands, Czechia, Brazil, Austria, Singapore, Portugal, Panama, Chile, Senegal, Morocco, Slovenia, New Zealand, Finland, Thailand, Uruguay, Argentina, Saudi Arabia, Romania, Greece, Mexico
Bovine serum albumin (BSA) is a common laboratory reagent derived from bovine blood plasma. It is a protein that serves as a stabilizer and blocking agent in various biochemical and immunological applications. BSA is widely used to maintain the activity and solubility of enzymes, proteins, and other biomolecules in experimental settings.
3
Streptavidin magnetic beads by Thermo Fisher Scientific
Sourced in United States, China, Germany
Streptavidin magnetic beads are a type of laboratory equipment used for various applications in biotechnology and molecular biology. They consist of magnetic particles coated with the protein streptavidin, which has a high affinity for the small molecule biotin. These beads can be used to capture and separate biotinylated molecules, such as proteins, nucleic acids, or cells, from complex samples.
4
Streptavidin hrp by Thermo Fisher Scientific
Sourced in United States, United Kingdom, China, Germany, France
Streptavidin-HRP is a conjugate of streptavidin, a protein that binds strongly to biotin, and horseradish peroxidase (HRP), an enzyme used as a reporter in various immunoassay and detection techniques. This product can be utilized in applications that require the specific interaction between streptavidin and biotin, such as ELISA, Western blotting, and immunohistochemistry.
5
Dynabead by Thermo Fisher Scientific
Sourced in United States, Norway, Germany, United Kingdom, France, China, Japan, Switzerland, Belgium, Canada, Italy, Australia
Dynabeads are magnetic beads used in various laboratory applications. They are designed to efficiently capture and isolate target molecules, such as proteins, nucleic acids, or cells, from complex samples. The magnetic properties of Dynabeads allow for easy separation and manipulation of the captured targets.
6
Dynabeads myone streptavidin c1 by Thermo Fisher Scientific
Sourced in United States
Dynabeads MyOne Streptavidin C1 are uniform, superparamagnetic polystyrene beads coated with streptavidin. They are designed for the efficient isolation and purification of biotinylated molecules, including proteins, nucleic acids, and cells.
7
Rneasy mini kit by Qiagen
Sourced in Germany, United States, United Kingdom, Netherlands, Spain, Japan, Canada, France, China, Australia, Italy, Switzerland, Sweden, Belgium, Denmark, India, Jamaica, Singapore, Poland, Lithuania, Brazil, New Zealand, Austria, Hong Kong, Portugal, Romania, Cameroon, Norway
The RNeasy Mini Kit is a laboratory equipment designed for the purification of total RNA from a variety of sample types, including animal cells, tissues, and other biological materials. The kit utilizes a silica-based membrane technology to selectively bind and isolate RNA molecules, allowing for efficient extraction and recovery of high-quality RNA.
8
Streptavidin beads by Thermo Fisher Scientific
Sourced in United States, China
Streptavidin beads are a type of solid-phase affinity matrix used for the separation and purification of biotinylated molecules. They consist of streptavidin, a protein derived from the bacterium Streptomyces avidinii, which binds to biotin with high affinity. These beads provide a convenient platform for the capture, separation, and immobilization of biotin-labeled proteins, nucleic acids, and other biomolecules.

More about "Streptavidin"

Streptavidin is a powerful biotechnology tool that has revolutionized research across various fields.
This tetrameric protein, derived from the bacterium Streptomyces avidinii, exhibits an exceptionally high affinity for the vitamin biotin, forming a robust non-covalent bond.
This unique property has made streptavidin an indispensable component in numerous experimental protocols, enabling the development of sensitive and specific assays, purification techniques, and labeling methods.
Streptavidin's versatility extends beyond its core functionality.
Streptavidin-conjugated reagents, such as Dynabeads M-280 Streptavidin and Dynabeads MyOne Streptavidin C1, provide researchers with valuable tools for the detection and localization of biotinylated molecules, including proteins, nucleic acids, and lipids.
These streptavidin-coated magnetic beads offer efficient capture and purification of target analytes, often in conjunction with techniques like affinity chromatography and immunoassays.
The stability and strength of the streptavidin-biotin interaction have also made it a popular choice for various applications, including floresscence microscopy and the RNeasy Mini Kit for RNA purification.
Additionally, the availability of streptavidin-HRP (horseradish peroxidase) conjugates enables sensitive detection and quantification of biotinylated molecules in diverse assays.
Bovine serum albumin (BSA) is often used in conjunction with streptavidin to block non-specific binding, enhancing the specificity and reliability of streptavidin-based experiments.
The combination of streptavidin's unparalleled affinity for biotin and the versatility of streptavidin-conjugated products has solidified its position as an indispensable tool in the world of biotechnology and biomedical research.