Alexa fluor 594 conjugated streptavidin

Manufactured by Thermo Fisher Scientific

Sourced in United States, Japan

Alexa Fluor 594-conjugated streptavidin is a fluorescent labeling reagent. It consists of the protein streptavidin coupled to the Alexa Fluor 594 dye. Streptavidin has a high affinity for biotin, enabling its use in various bioanalytical techniques.

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Lab products found in correlation

B 1355, by Vector Laboratories (5 mentions) Vectashield medium h 1400, by Vector Laboratories (3 mentions) Triton x 100, by Merck Group (2 mentions) Gelfoam, by Pfizer (2 mentions) Vectashield, by Vector Laboratories (2 mentions) Hoechst 33258, by Thermo Fisher Scientific (2 mentions) Chicken polyclonal primary antibody to beta galactosidase, by Abcam (2 mentions) Biotinylated goat anti chicken secondary antibody, by Jackson ImmunoResearch (2 mentions) Paraformaldehyde (pfa), by Merck Group (2 mentions) Las x software, by Leica (2 mentions) Prolong gold antifade mounting media, by Thermo Fisher Scientific (2 mentions) Tcs spe confocal microscope, by Leica (2 mentions) Cryostar nx70 cryostat, by Thermo Fisher Scientific (2 mentions) Gl7 alexa fluor 488, by Thermo Fisher Scientific (2 mentions) Ki 67 horizon v450, by BD (2 mentions) Cryomatrix embedding resin, by Thermo Fisher Scientific (2 mentions) Anti b220 biotin, by BD (2 mentions) Texas red conjugated goat anti rabbit, by Vector Laboratories (2 mentions) Alexa fluor 647 conjugated streptavidin, by Thermo Fisher Scientific (2 mentions) Biotin xx tsa kit 21, by Thermo Fisher Scientific (2 mentions)

Spelling variants of this product

Alexa Fluor 594 (2045 citations) Alexa 594 (538 citations) Alexa Fluors (21 citations) Streptavidin Alexa 594 (17 citations) Alexa 594-conjugated streptavidin (8 citations) Alexa Fluor-conjugated streptavidin (3 citations) Alexa Fluor 594-conjugated streptavidin (S11227; (2 citations)

42 protocols using alexa fluor 594 conjugated streptavidin

Immunostaining of Wisteria floribunda Lectin

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The sections were treated with 0.1% Triton X-100 and PBS at room temperature for 15 min. After three washes with PBS, sections were incubated with 10% normal goat serum (ImmunoBioScience Corp., Mukilteo, WA, USA) in PBS at room temperature for 1 h. They were then washed thrice with PBS and incubated overnight at 4 °C in PBS containing biotinylated Wisteria floribunda agglutinin (WFA) (B-1355, Vector Laboratories; 1:200) and/or antibodies described in the subsequent text. After washing with PBS, the sections were incubated with Alexa Fluor 594-conjugated streptavidin (S11227; Molecular Probes, Eugene, OR) and/or the corresponding secondary antibodies (described in the subsection Antibodies and lectins) at room temperature for 2 h. The labeled sections were then rinsed with PBS and mounted on glass slides using Vectashield medium (H-1400; Vector Laboratories, Funakoshi Co., Tokyo, Japan). The prepared slides were stored at 4 °C until microscopic analysis was performed.

Ueno H., Takahashi Y., Murakami S., Wani K., Matsumoto Y., Okamoto M, & Ishihara T. (2022). Fingolimod increases parvalbumin-positive neurons in adult mice. IBRO Neuroscience Reports, 13, 96-106.

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Immunocytochemistry of Actin and Nuclei

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Coverslips with fixed MEF cells were prepared, washed in PBS, and blocked with 1% BSA/PBS for 5 min. Following a PBS wash, the nuclei were stained using Hoechst (Life Technologies Molecular Probes, Grand Island, NY) 1:100 in H₂O for 30 min at 37°C. Cells were then blocked using a casein block solution (Scytek) for 5 min, AVIDIN/Biotin blocks (VECTOR) for 20 min each at room temperature (RT) followed by a 5 min PBS wash. Biotinylated p5+14 or CGGY-p5G (control) at 1.6 μg/mL in PBS was added and incubated overnight at 4°C. Following a PBS wash Alexa Fluor 594-conjugated streptavidin (Molecular Probes) was added at a 1:200 dilution in PBS for 1h at RT. Cells were then permeabilized with 0.2% Triton X-100 (Sigma) in PBS for 10 min at RT and washed with a solution of 1% BSA in PBS for 30 min. The cells were then stained with Alexa Fluor 488-conjugated phalloidin (Molecular Probes) at a 1:100 dilution of stock in 1% BSA/PBS, for 45 min at RT to visualize actin filaments. Slides were cover-slipped using a fluorescent mounting medium (Dako) to minimize photobleaching.

Dogra P., Martin E.B., Williams A., Richardson R.L., Foster J.S., Hackenback N., Kennel S.J., Sparer T.E, & Wall J.S. (2015). Novel Heparan Sulfate-Binding Peptides for Blocking Herpesvirus Entry. PLoS ONE, 10(5), e0126239.

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Spinal Cord Tissue Cryosectioning and Axon Labeling

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Sixteen-micrometer cross cryosections of paraformaldehyde-fixed SC tissues were used. Tissue sections were imaged using a Zeiss LSM 800 confocal microscope. For Gfap staining, mouse anti-Gfap (ZIRC, Zrf1, AB_10013806, 1:1000) and Alexa Fluor-488 anti-mouse secondary antibody (Invitrogen, 1:200) were used. For anterograde axon labeling, zebrafish were anesthetized using MS-222 and fine scissors were used to transect the spinal cord 4 mm rostral to the lesion site. Biocytin-soaked Gelfoam Gelatin Sponge was applied at the new injury site (Gelfoam, Pfizer, cat# 09-0315-08; Biocytin, saturated solution, Sigma, cat# B4261). Fish were euthanized 4 hours post-treatment and Biocytin was histologically detected using Alexa Fluor 594-conjugated Streptavidin (Molecular Probes, cat# S-11227).

Klatt Shaw D, & Mokalled M.H. (2021). Efficient CRISPR/Cas9 mutagenesis for neurobehavioral screening in adult zebrafish. G3: Genes|Genomes|Genetics, 11(8), jkab089.

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Immunostaining of Extracellular Matrix Components

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The sections were then treated with 0.1% Triton X-100 and PBS at room temperature for 15 min. After three washes with PBS, sections were incubated with 10% normal goat serum (ImmunoBioScience Corp., Mukilteo, WA, USA) in PBS at room temperature for 1 h. They were then washed thrice with PBS and incubated overnight at 4 °C in PBS containing biotinylated WFA (B-1355, Vector Laboratories; 1:200) and the antibodies described in the subsequent text. Subsequently, the sections were incubated with Alexa Fluor 594-conjugated streptavidin (S11227; Molecular Probes, Eugene, OR, USA) and the corresponding secondary antibodies (described in the Lectins and antibodies subsection) at room temperature for 2 h. The labeled sections were then rinsed with PBS and mounted on glass slides using a Vectashield medium (H-1400; Vector Laboratories, Funakoshi Co., Tokyo, Japan). The prepared slides were stored at 4 °C until further microscopic analyses.

Ueno H., Takahashi Y., Murakami S., Wani K., Miyazaki T., Matsumoto Y., Okamoto M, & Ishihara T. (2023). Component-specific reduction in perineuronal nets in senescence-accelerated mouse strains. IBRO Neuroscience Reports, 14, 111-121.

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Anterograde Axon Tracing in Zebrafish

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Anterograde axon tracing was performed on adult fish at 4 wpi. Fish were anaesthetized using MS-222 and fine scissors were used to transect the cord 4 mm rostral to the lesion site. Biocytin-soaked Gelfoam Gelatin Sponge was applied at the new injury site (Gelfoam, Pfizer, cat# 09–0315-08; Biocytin, saturated solution, Sigma, cat# B4261). Fish were euthanized 6 hours post-treatment and Biocytin was histologically detected using Alexa Fluor 594-conjugated Streptavidin (Molecular Probes, cat# S-11227).

Klatt Shaw D., Muraleedharan Saraswathy V., Zhou L., McAdow A.R., Burris B., Butka E., Morris S.A., Dietmann S, & Mokalled M.H. (2021). Localized EMT reprograms glial progenitors to promote spinal cord repair. Developmental cell, 56(5), 613-626.e7.

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Immunolabeling of Cryostat Sections

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We treated the cryostat sections with 0.1% Triton X-100 with PBS at room temperature for 15 min. After three washes with PBS, we incubated the sections with 10% normal goat serum (ImmunoBioScience Corp., Mukilteo, WA) in PBS at room temperature for 1 h, we washed them three times with PBS, and incubated them overnight at 4 °C in PBS containing biotinylated WFA (B-1355, Vector Laboratories; 1:200) and the antibodies described in the subsection Antibodies and Lectins. After washing with PBS, we incubated the sections with the corresponding secondary antibodies (described in the subsection Antibodies and lectins) and Alexa Fluor 594-conjugated streptavidin (S11227; Molecular Probes, Eugene, OR) at room temperature for 2 h. We rinsed the labeled sections again with PBS and we mounted them on glass slides with Vectashield medium (H-1400; Vector Laboratories, Funakoshi Co., Tokyo, Japan). We stored the prepared slides at 4 °C until we used them in the microscopy analysis.

Ueno H., Suemitsu S., Murakami S., Kitamura N., Wani K., Matsumoto Y., Okamoto M, & Ishihara T. (2018). Layer-specific expression of extracellular matrix molecules in the mouse somatosensory and piriform cortices. IBRO Reports, 6, 1-17.

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Quantifying Cellular DNA Replication

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Cells were treated with specified doses of hydroxyurea (Sigma) for
24 hr before incubation with 10 mM EdU (CarboSynth) for 20 min. Cells were
fixed with formalin, permeabilized with 1% Triton X-100 (Sigma), before
performing a click reaction to biotinylate incorporated EdU in 10 mM sodium
ascorbate (Sigma), 10 μM Azide-PEG3-biotin (Sigma), and 2 mM
CuSO₄ (Ricca) for 20 min. After washing, cells were blocked
in 1% BSA and then labeled with AlexaFluor 594-conjugated Streptavidin
(Molecular Probes). Images were collected on a Nikon Eclipse TE200
microscope and quantified using MATLAB.

Fang Y., McGrail D.J., Sun C., Labrie M., Chen X., Zhang D., Ju Z., Vellano C.P., Lu Y., Li Y., Jeong K.J., Ding Z., Liang J., Wang S.W., Dai H., Lee S., Sahni N., Mercado-Uribe I., Kim T.B., Chen K., Lin S.Y., Peng G., Westin S.N., Liu J., O’Connor M.J., Yap T.A, & Mills G.B. (2019). Sequential therapy with PARP and WEE1 inhibitors minimizes toxicity while maintaining efficacy. Cancer cell, 35(6), 851-867.e7.

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Quantifying HLA-G Expression on NK Cells

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Expression of the anti-HLA-G fragment on the NK cell surface was determined using protein L staining, as described previously.34 (link) Briefly, HLA-G CAR-transduced primary NK cells were harvested, washed, resuspended in PBS containing 4% FBS and protein L (1 µg), incubated for 45 min, washed three times, resuspended for 45 min in PBS containing Alexa Fluor 594-conjugated streptavidin (5 µg/mL, Thermo Fisher Scientific) and FITC-conjugated anti-CD56 antibody, and washed again three times. Protein L and CD56 dual-positive cells were examined by flow cytometry, with parental NK cells used as background controls.

Jan C.I., Huang S.W., Canoll P., Bruce J.N., Lin Y.C., Pan C.M., Lu H.M., Chiu S.C, & Cho D.Y. (2021). Targeting human leukocyte antigen G with chimeric antigen receptors of natural killer cells convert immunosuppression to ablate solid tumors. Journal for Immunotherapy of Cancer, 9(10), e003050.

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Pneumococcal Capsule Visualization Protocol

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A frozen aliquot of pneumococcus was thawed, washed by centrifugation, and re-suspended with 1 ml of HBC. Then, 250 μl of this suspension was mixed with 5 μl of biotin-conjugated DBA or HPA (50 μg/ml final concentration) and incubated for 30 min on ice. After washing with HBC, the bacteria were incubated with 2 μg/ml Alexa Fluor-594-conjugated streptavidin (Thermo Fisher Scientific, Rockford, IL) for 30 min on ice. To visualize the capsule of D39 and SPEC6B, the bacteria were also incubated with Hyp2M2 and Hyp6BM1, respectively, and visualized with Alexa Fluor-488-conjugated goat anti-mouse IgM [23 (link)]. Bacteria were washed and re-suspended in PBS with 5 μM DRAQ5 (Thermo Fisher Scientific) and incubated in the dark for 15 min at room temperature (RT). After washing twice with PBS, the bacterial pellet was re-suspended in 10% neutral buffered formalin (Thermo Fisher Scientific) and transferred to a cover slip to dry for 15 min at RT. The coverslip was washed three times with PBS, allowed to dry, and then placed on a slide with 20 μl of Fluromount G (Thermo Fisher Scientific). After a 1-hour incubation at RT, bacteria were examined with either confocal or structured illumination microscopy (Nikon A1R), using a 100X oil objective.

Zhou M.L., Frost M.R., Xu Y.C, & Nahm M.H. (2020). Phosphorylcholine esterase is critical for Dolichos biflorus and Helix pomatia agglutinin binding to pneumococcal teichoic acid. Journal of basic microbiology, 60(10), 905-915.

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In Vivo Biotinylation of Skin

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The assay was performed according to a previously described method [10 (link)]. A volume of 50 μL of 10 mg/mL EZ-Link™ Sulfo-NHS-LC-Biotin (Thermo Fisher Scientific) in phosphate-buffered saline containing 1 mM CaCl₂ was injected into the dermis of the back skin. After 30 min, skin samples were collected and frozen in liquid nitrogen. Frozen sections were fixed and soaked in blocking solution as described for immunohistochemistry. Alexa Fluor 594-conjugated streptavidin (1:10,000 dilution; Thermo Fisher Scientific) was incubated with secondary antibodies.

Umehara Y., Trujillo-Paez J.V., Yue H., Peng G., Nguyen H.L., Okumura K., Ogawa H, & Niyonsaba F. (2023). Calcitriol, an Active Form of Vitamin D3, Mitigates Skin Barrier Dysfunction in Atopic Dermatitis NC/Nga Mice. International Journal of Molecular Sciences, 24(11), 9347.

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