Alexa fluor 488 or 568

Manufactured by Thermo Fisher Scientific

Sourced in United States, France, Japan

Alexa Fluor 488 and Alexa Fluor 568 are fluorescent dyes used in various biological applications, including cell imaging, flow cytometry, and immunoassays. These dyes are excited by specific wavelengths of light and emit light at different wavelengths, allowing them to be detected and used as labels for target molecules or cells. The core function of these dyes is to enable the detection and visualization of labeled targets through fluorescence-based techniques.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Merck Group (7 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (4 mentions) Lsm 710 confocal microscope, by Zeiss (4 mentions) Lsm 700, by Zeiss (4 mentions) Vectashield, by Vector Laboratories (4 mentions) Axiovert 200m, by Zeiss (4 mentions) Bx51 microscope, by Olympus (3 mentions) Bovine serum albumin (bsa), by Merck Group (3 mentions) Prolong gold antifade reagent, by Thermo Fisher Scientific (3 mentions) Lsm 780 confocal microscope, by Zeiss (3 mentions) Mouse anti oct3 4, by Santa Cruz Biotechnology (3 mentions) Zen software, by Zeiss (3 mentions) Mounting media, by Agilent Technologies (2 mentions) Poly l lysine (pll), by Merck Group (2 mentions) Prolong gold, by Thermo Fisher Scientific (2 mentions) Normal donkey serum (nds), by Thermo Fisher Scientific (2 mentions) Eclipse e800, by Nikon (2 mentions) Apotome, by Zeiss (2 mentions) Triton x 100, by Thermo Fisher Scientific (2 mentions) Ve cadherin, by Santa Cruz Biotechnology (2 mentions)

Close spelling variants of this product

Alexa Fluor 488 or Alexa Fluor 568 Alexa Fluor 488 or 568 secondary antibodies Anti-mouse IgG Alexa Fluor 488 or 568 Alexa Fluor 488 or 568-labeled secondary antibodies Alexa Fluor 488 or 568 phalloidin Alexa Fluor™ 488 or 568 Tyramide Reagent Alexa Fluor 488- or 568-labeled IgG Alexa Fluor 488- or 568-conjugated donkey anti-species Alexa Fluor 488 or Alexa Fluor 568 phalloidin IgG Alexa Fluor 488 or 568-conjugated goat anti-mouse or anti-rabbit antibodies

105 protocols using alexa fluor 488 or 568

Molecular Profiling of Inner Ear Development

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The inner ear was fixed in 4% para-formaldehyde (PFA) in phosphate-buffered saline (PBS) overnight at 4°C, then the cochlea and vestibule were dissected in DEPC-treated PBS. In-situ hybridisation was performed using Dig-labelled ATOH1 riboprobe (gift from M. A. Basson, Craniofacial Development and Stem Cell Biology, King's College London, London, UK). Antibodies used were: rabbit MYO7A (1:1000, Proteus), rabbit acetylated α-tubulin (1:500, Abnova), rabbit p75^NTR (1:500, Abnova), detected with anti-rabbit Alexa Fluor 488 or 568 (1:1000, Invitrogen), mouse NF-M (1:100, Invitrogen), detected with anti-mouse Alexa Fluor 488 or 568 (1:1000, Invitrogen) and Alexa Fluor 488 or 568 Phalloidin (1:1000, Invitrogen).

Ahmed M., Ura K, & Streit A. (2015). Auditory hair cell defects as potential cause for sensorineural deafness in Wolf-Hirschhorn syndrome. Disease Models & Mechanisms, 8(9), 1027-1035.

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Immunofluorescence Staining of Cells

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Poly-L-lysine cover glasses were used for seeding the cells as previously described (Bhadra et al., 2015 (link)). Blocking was performed using 1% BSA for 1 h at room temperature. The cover slips were incubated with the primary antibody (see above) overnight at 4°C (in a covered chamber surrounded with damp Kim wipes). Cells were incubated with the secondary antibody for 1 h at room temperature (protected from light). The secondary antibodies used were as follows: anti-rabbit Alexa Fluor 488 or 568 (1:200, Invitrogen), anti-mouse Alexa Fluor 488 or 568 (1:200, Invitrogen). DAPI (1:1000, MP Biomedicals, LLC, Santa Ana, California, USA) labels the nucleus. Cells were mounted with Vectashield (Vector Laboratories) and examined with an Eclipse TE2000-U (Nikon, Melville, NY, USA) at 40× or 60×.

Banerji R., Skibbens R.V, & Iovine M.K. (2017). Cohesin mediates Esco2-dependent transcriptional regulation in a zebrafish regenerating fin model of Roberts Syndrome. Biology Open, 6(12), 1802-1813.

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Immunofluorescence Imaging of Transfected Cells

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HeLa cells and Raw264.7 stable cells were seeded onto glass coverslips. After 24 h, cells were transfected using Lipofectamine 2000 (Invitrogen). Cells were washed 18 h after transfection, fixed in 4% paraformaldehyde, permeabilized with 0.5% Triton X-100 and blocked with 5% BSA for 1 h. Cells were stained with the designated primary antibodies, followed by labelling with anti-mouse Alexa Fluor 568 or 488 and anti-rabbit Alexa Fluor 568 or 488 (Molecular Probes). After antibody labelling, cells were counterstained with Hoechst (33342, Molecular Probes). Cells were imaged with a confocal microscope (Nikon) and were processed with NIS-elements confocal (Nikon).

Choi Y., Park S.J., Sun Y., Yoo J.S., Pudupakam R.S., Foo S.S., Shin W.J., Chen S.B., Tsichlis P.N., Lee W.J., Lee J.S., Li W., Brennan B., Choi Y.K, & Jung J.U. (2019). Severe fever with thrombocytopenia syndrome phlebovirus non-structural protein activates TPL2 signalling pathway for viral immunopathogenesis. Nature microbiology, 4(3), 429-437.

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Immunofluorescence Microscopy of Cells

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Cells were cultured and transfected on Lab-Tek II chamber slides. Cells were fixed for 15 min in 4% formaldehyde, washed with phosphate buffered saline (PBS) 3 times, and incubated in blocking buffer (PBS, 5% goat serum, 0.1% saponin) for 1h at room temperature. Primary antibody was diluted in antibody solution (PBS, 1% BSA, 0.1% saponin) and incubated at 4C overnight. After washed with PBS 3 times, cells were incubated with secondary antibody conjugated with Alexa Fluor 488 or 568 (Invitrogen) for 1h at room temperature. After PBS wash, the slides were mounted with Prolong Gold Antifade reagent. The immunofluorescent pictures were taken by the Zeiss LSM510 Meta Confocal Laser Scanning Microscope or Nikon Eclipse E800, and analyzed by the LSM image browser or Nikon NIS Elements BR 3.0 software.

Chen L.X., Morales-Alcala C.C, & Riobo-Del Galdo N.A. (2018). Autophagic Flux is Regulated by Interaction Between the C-terminal Domain of Patched1 and ATG101. Molecular cancer research : MCR, 16(5), 909-919.

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Immunofluorescence Visualization of STAT1/STAT3

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Immunofluorescence was performed according to the manual. Cells were grown on cover slips coated with poly-l-lysine (Sigma Aldrich) in a six-well plate and fixed with 3% para-formaldehyde in PBS (pH 7.4). Cells were rinsed three times with PBS, permeabilized with Triton X-100, washed again with PBS, and incubated with 200 µL of anti-STAT1 and anti-STAT3 antibodies (1:50, Sigma Aldrich) overnight at room temperature in a humidified chamber. Cover slips were rinsed three times in PBS and incubated with secondary antibody conjugated with Alexa Fluor 488 or 568 (Invitrogen) at a 1:250 dilution for 1 hour at room temperature. After three rinses in PBS, coverslips were mounted on a slide using mounting media (Dako, Carpinteria, CA, USA). Cells were visualized with a Zeiss LSM 710 confocal microscope (Carl-Zeiss Company, Jena, Germany).

Liu Z., Zhang Y., Chen Y., Lin Y., Lin Z, & Wang H. (2018). STAT1 inhibits STAT3 activation in esophageal squamous cell carcinoma. Cancer Management and Research, 10, 6517-6523.

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Immunofluorescence Microscopy of Mastigonemes

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For immunofluorescence microscopy of mastigonemes, cells were fixed for 30 min at RT with freshly prepared 3.7% formaldehyde, allowed to settle onto poly-l-lysine multiwell slides for 5 min, washed three times with PBS for 10 min each, and air dried. The specimens were treated with 0.5% Triton X-100 in PBS for 5 min, blocked in PBS with 1% BSA and 0.05% Tween20, and incubated in anti-MST1 (1:500; Nakamura et al., 1996 (link)) overnight at 4°C in blocking buffer. After incubation in the secondary antibodies (anti-mouse IgG conjugated to Alexa Fluor 488 or 568; 1:1,000; Invitrogen; 90–120 min; RT) specimens were mounted with ProlongGold (Invitrogen). An Axio Imager.Z1 equipped with a 100×/1.3-NA EC Plan-Neofluar objective, an ApoTome, an axiocam MRm camera, and Axiovision (Zeiss) were used for imaging and image acquisition. To detect PKD2-GFP, we used anti-GFP 1:1,000 (mouse; Sigma-Aldrich; G 6539). Image brightness and contrast were adjusted using Photoshop (CC2018; Adobe), and figures were assembled using Illustrator (CC2018; Adobe). Acquisition conditions and adjustments of brightness and contrast were similar for images mounted together.

Liu P., Lou X., Wingfield J.L., Lin J., Nicastro D, & Lechtreck K. (2020). Chlamydomonas PKD2 organizes mastigonemes, hair-like glycoprotein polymers on cilia. The Journal of Cell Biology, 219(6), e202001122.

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Histological Analysis of Bone Remodeling

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Fixed samples were decalcified in 20% EDTA (pH 7.5), embedded in paraffin and cut into 5 μm sections using an HM360 microtome (Microm). Hematoxylin (VWR) and eosin (Sigma–Aldrich) staining was performed to assess histomorphology. Tartrate-resistant acid phosphatase (TRAP) (Sigma) was used to detect osteoclasts according to the manufacturers’ protocols. For immunofluorescence staining, slides were subjected to sodium citrate buffer at 95 °C for 20 min for antigen retrieval, permeabilized with 0.5% Triton X-100 (Beyotime) for 10 min, and blocked with 5% BSA for 1 h. Slides were incubated with Anti-Runx2 (1:200, Abcam, ab23981), Anti-GFP (1:50, Santa Cruz, sc-9996), Anti-RFP (1:50, Santa Cruz, sc-390909), Anti-Klotho (1:100, R&D, AF1819), Anti-Klotho (1:100, Santa Cruz, sc-515939), or Anti-Rankl (1:100, R&D, AF462) antibody overnight at 4 °C, and a fluorescence-conjugated secondary antibody, Alexa Fluor 488 or 568 (Invitrogen, 1:1000) for 1 h at room temperature. Nuclei were counterstained with DAPI (Vector). Images were captured on an Olympus confocal microscope FV3000 (Olympus).

Fan Y., Cui C., Rosen C.J., Sato T., Xu R., Li P., Wei X., Bi R., Yuan Q, & Zhou C. (2022). Klotho in Osx+-mesenchymal progenitors exerts pro-osteogenic and anti-inflammatory effects during mandibular alveolar bone formation and repair. Signal Transduction and Targeted Therapy, 7, 155.

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Immunofluorescence Analysis of GluA4 and Calbindin in WT and GluA4-KO Mouse Brains

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Brain tissues of three WT and two GluA4-KO mice were processed for immunofluorescence. Mice were deeply anesthetized with ketamine and xylazine (100 mg/kg each) and transcardially perfused with 4% paraformaldehyde in 0.1 M phosphate buffer (PB) pH 7.2. Brains were post-fixed overnight in the same fixative and cryoprotected with 30% sucrose overnight, and then cryostat sectioned at a thickness of 30 μm. Sections were rinsed with PB and then incubated in blocking buffer, containing 10% normal goat serum, 1% BSA and 0.3% triton X-100 in PB, for 1 hour at room temperature. The primary antibodies, rabbit anti-GluA4 (2 μg/mL; Chemicon AB1508) or guinea pig anti-Calbindin (1:500; Synaptic Systems 214 004), were added to the sections and incubated overnight at 4°C in blocking buffer. After rinsing well with blocking buffer, the sections were incubated with anti-species specific secondary antibodies, conjugated with Alexa Fluor® 488 or 568 (Invitrogen, Carlsbad, CA), in blocking buffer for 1 hour at room temperature. Nuclei were visualized using Dapi. Images of immunostained sections were captured on an Olympus BX51 microscope equipped with epi-fluorescence illumination (100W mercury burner) and CellSensDimension software for image acquisition (Olympus Corporation, Tokyo, Japan).

García-Hernández S, & Rubio M.E. (2021). Role of GluA4 in the acoustic and tactile startle responses. Hearing research, 414, 108410.

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Subcellular Localization of NHE7 and Rab Proteins

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Cells were seeded onto a coverslip, and fixed with 4% paraformaldehyde. After rinsing, the cells were permeabilized with 0.1% Triton X‐100 (#22686, USB Corporation, Cleveland, OH, USA) before incubating with blocking buffer (5% BSA in PBS). Primary antibodies against NHE7 (1:100; #PA5‐61843, Invitrogen), early endosome antigen 1 (1:100; #ab70521, EEA1, Abcam), Rab7 (1:100; #95746, Cell Signaling Technology, Danvers, MA, USA), Rab11a (1:100; #STJ140068, St John's Laboratory, London, UK), Rab21 (1:100; #sc‐81917, Santa Cruz Biotechnology), or Golgin‐97 (1:100; #A21270, Invitrogen) was used to treat the cells. After rinsing in PBS, cells were maintained in secondary antibodies tagged with Alexa Fluor™ 488 or 568 (Invitrogen). Cell nuclei were stained with DAPI, and coverslips were mounted onto slides (Vector Laboratories, Burlingame, CA, USA). An LSM900 confocal microscope was used to capture images.

Yao Y., Xu Y., Yu L., Xue T., Xiao Z., Tin P., Fung H., Ma H., Yun J, & Yam J.W. (2023). NHE7 upregulation potentiates the uptake of small extracellular vesicles by enhancing maturation of macropinosomes in hepatocellular carcinoma. Cancer Communications, 44(2), 251-272.

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Immunofluorescence Staining of Kidney Tissue

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Immunofluorescence staining was performed by following the method described previously [53 (link)]. Briefly, kidneys were embedded in optimal cutting temperature compound (Sakura Finetek, Tokyo, Japan), and frozen in liquid nitrogen. Sections of 5-μm thickness were cut by a cryostat (CM3050; Leica, Wetzlar, Germany); then incubated with primary antibodies against type I collagen (Acris Antibodies, Germany), type IV collagen, type V collagen, synaptopodin, VE-cadherin (Santa Cruz Biotechnology, Santa Cruz, CA, USA), nephrin, and podocalyxin (R&D Systems, Minneapolis, MN, USA), respectively, at 4 °C overnight, and followed by secondary antibodies conjugated to Alexa Fluor^® 488 or 568 (Invitrogen, Carlsbad, CA, USA); double-stained with rhodamine-conjugated phalloidin (Life Technologies, Gaithersburg, MD, USA) for F-actin; and finally submerged in fluoroshield mounting medium containing 4′,6-diamidino-2-phenylindole (DAPI) (Abcam, Cambridge, UK). Confocal imaging was performed according to the method described previously [26 (link)] with a confocal microscope (LSM700; Carl Zeiss, Jena, Germany). Alexa Fluor^® 488, and 568 signals were detected at laser excitation wavelengths of 488 nm and 543 nm, respectively.

Chang C.J., Wang P.C., Huang T.C, & Taniguchi A. (2019). Change in Renal Glomerular Collagens and Glomerular Filtration Barrier-Related Proteins in a Dextran Sulfate Sodium-Induced Colitis Mouse Model. International Journal of Molecular Sciences, 20(6), 1458.

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