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Alexa fluor 647 anti mouse

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Alexa Fluor 647 anti-mouse is a fluorescently labeled secondary antibody produced by Jackson ImmunoResearch. It is designed to detect and bind to mouse primary antibodies, allowing for visualization and detection in various immunoassays and imaging techniques.

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Lab products found in correlation

Anti rabbit cy3, by Jackson ImmunoResearch (3 mentions) Alexa fluor 488 anti rabbit, by Jackson ImmunoResearch (3 mentions) Alexa fluor 647 anti rabbit, by Jackson ImmunoResearch (3 mentions) Goat anti sycp3, by Santa Cruz Biotechnology (2 mentions) Mouse monoclonal anti mlh1 4c9c7, by Cell Signaling Technology (2 mentions) Amca anti goat, by Jackson ImmunoResearch (2 mentions) Facsaria, by BD (2 mentions) Alexa fluor 488 anti mouse, by Jackson ImmunoResearch (2 mentions) Anti α actinin, by Abcam (2 mentions) Anti tuj1, by Abcam (2 mentions) Anti ctnt, by Thermo Fisher Scientific (2 mentions) Anti alb, by Abcam (2 mentions) Ab13524, by Abcam (1 mentions) Ab24170, by Abcam (1 mentions) G3bp1, by BD (1 mentions) Alexa fluor 647 anti rabbit igg, by Thermo Fisher Scientific (1 mentions) α tubulin, by Santa Cruz Biotechnology (1 mentions) Stat3, by Cell Signaling Technology (1 mentions) Advanced rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin solution, by Thermo Fisher Scientific (1 mentions)

12 protocols using alexa fluor 647 anti mouse

1

Immunostaining of Mouse Spermatocyte Chromosomes

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Surface-spread chromosomes of mouse spermatocytes were prepared from 18 days postpartum male mice using the drying-down technique as previously described33 (link). Immuno-staining was performed as previously described33 (link) using the following primary and secondary antibodies: goat anti-SYCP3 (Santa Cruz Biotechnology, sc-20845; 1:200), rabbit anti-MSH4 (a gift from Dr. Paula Cohen; 1:200), mouse monoclonal anti-MLH1 (4C9C7) (Cell Signaling Technology, 3515S; 1:30), AMCA anti-goat (Jackson ImmunoResearch, 705–155-147; 1:50), Cy3 anti-rabbit (Jackson ImmunoResearch, 711–165-152; 1:100), and AlexaFluor647 anti-mouse (Jackson ImmunoResearch, 715–605-151; 1:100).
Kulkarni D.S., Owens S.N., Honda M., Ito M., Yang Y., Corrigan M.W., Chen L., Quan A.L, & Hunter N. (2020). PCNA activates the MutLγ endonuclease to promote meiotic crossing over. Nature, 586(7830), 623-627.
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2

Immunofluorescence Labeling of Cellular Markers

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Primary antibodies against the following antigens were purchased from the sources specified and used at the dilutions indicated: TFEB (Sigma-Aldrich, SAB4503154; diluted 1:500), LAMP1 (Abcam, ab24170; 1:1000). LAMP2 (Abcam, ab13524; 1:500), Nrf2 (Abcam, ab31163; 1:200), G3BP1 (BD Biosciences; 1:2000), HuR (Santa Cruz Biotechnology, sc-5261; 1:2000), importin-α1 (Santa Cruz Biotechnology, sc-6917; 1:500), α-tubulin (Santa Cruz Biotechnology, sc-5286; 1:500), STAT3 (Cell Signaling Technology, #9139; 1:1250). Secondary antibodies: AlexaFluor®647 anti-rabbit IgG (Life Technologies, A21244; 1:500) and AlexaFluor®647 anti-rat IgG (Life Technologies, A21247; 1:500), AlexaFluor®647 anti-mouse (Jackson ImmunoResearch, 715-605-150; 1:400), Cy3™-anti-rabbit (Jackson ImmunoResearch, 711-165-152; 1:250).
Maysinger D., Gran E.R., Bertorelle F., Fakhouri H., Antoine R., Kaul E.S., Samhadaneh D.M, & Stochaj U. (2020). Gold nanoclusters elicit homeostatic perturbations in glioblastoma cells and adaptive changes of lysosomes. Theranostics, 10(4), 1633-1648.
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3

Isolation and Characterization of Ductal and Acinar Cells

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Ductal and acinar cells from fresh tissue were isolated as previously described using HPx1 (acinar) and CD133 (ductal) markers (74 (link)).
To obtain ductal and de-differentiated acinar cells from in vitro cultures exocrine preparations were cultured as previously described (59 ,70 (link)) in Advanced RPMI 1640 medium containing 5% heat-inactivated fetal bovine serum and 1% penicillin-streptomycin solution (Life Technologies) under 5% CO2 atmosphere at 37 °C. FITC-conjugated UEA-1 (Ulex Europeaus Agglutinin-1, Sigma, Overijse, Belgium) lectin labeling of acinar cells was performed according to Houbracken et al. and Baldan et al. (59 ,70 (link)). Exocrine cell fraction labeled with UEA-1 was kept in suspension culture. At day 4 of culture, cell clusters were dissociated following the protocol of Baldan et al. (59 ). Cells were stained with carbohydrate antigen 19.9 (mouse monoclonal anti-human CA19.9, Dako, Heverlee, Belgium). Alexa Fluor 647 anti-mouse (Jackson Laboratory, Westgrove, PA, USA) was used as secondary antibody. Analysis and cell sorting were performed on a BD FACSAria (BD Biosciences). Viable, single cells were gated based on forward and side scatter.
Espinet E., Gu Z., Imbusch C.D., Giese N.A., Büscher M., Safavi M., Weisenburger S., Klein C., Vogel V., Falcone M., Insua-Rodríguez J., Reitberger M., Thiel V., Kossi S.O., Muckenhuber A., Sarai K., Lee A.Y., Backx E., Zarei S., Gaida M.M., Rodríguez-Paredes M., Donato E., Yen H.Y., Eils R., Schlesner M., Pfarr N., Hackert T., Plass C., Brors B., Steiger K., Weichenhan D., Arda H.E., Rooman I., Kopp J.L., Strobel O., Weichert W., Sprick M.R, & Trumpp A. (2020). Aggressive PDACs show hypomethylation of repetitive elements and the execution of an intrinsic IFN program linked to a ductal cell-of-origin. Cancer discovery, 11(3), 638-659.
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4

Isolation and Analysis of CA19.9+ Cells

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UEA1-FITC conjugated exocrine cell fraction was kept in suspension culture. At day 4, cell clusters were dissociated using StemPro™ Accutase™ Cell Dissociation Reagent (ThermoFisher Scientific, Waltham, Massachusetts, USA) following the protocol for dissociation of neurospheres, filtered over a 40 µm filter and cells were incubated with mouse monoclonal anti-human carbohydrate antigen 19.9 antibody (anti-CA19.9, Dako, Heverlee, Belgium; 2 µl per million cells in 200 µl) or isotype control (IgG1 kappa, Abcam, Cambridge, MA, USA; 2 µl per million cells in 200 µl) for 15 minutes at 4 °C. Cells were washed with FBS buffer (PBS + 3% FBS) and incubated for 15 minutes at 4 °C with secondary antibody Alexa fluor 647 anti-mouse (Jackson Laboratory, Westgrove, PA, USA; 2 µl per million cells in 200 µl). Analysis and cell sorting was performed on a BD FACSAria (BD Biosciences, Erembodegem, Belgium). Viable, single cells were gated based on forward and side scatter. After sort, cells were immediately collected in RLT buffer (Qiagen, Germantown, MD 20874, USA) and kept on ice. RNA extraction was immediately performed after sorting.
Baldan J., Houbracken I., Rooman I, & Bouwens L. (2019). Adult human pancreatic acinar cells dedifferentiate into an embryonic progenitor-like state in 3D suspension culture. Scientific Reports, 9, 4040.
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5

Immunofluorescence Staining Protocol for Cell Characterization

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Cells were fixed with 4% PFA in 1X DPBS for 10 minutes, washed twice 1X PBS, permeabilized for 20 minutes in 0.1% Triton X-100 1X PBS, washed twice with 1X PBS, blocked for at least 60 minutes in 5% BSA 1X PBS, stained overnight at 4°C with primary antibody diluted in 5% BSA 1X PBS, washed three times with 1X PBS, stained for 60 minutes with secondary antibody diluted in 5% BSA 1X PBS, and lastly washed three times with 1X PBS before imaging. Primary antibodies and relevant dilutions were as follows: anti-Tuj1, 1:500 (Abcam, # ab7751); anti-Alb, 1:500 (Abcam, ab207327); anti-cTnT, 1:400 (Thermo Fisher, # MA5-12960); anti-αActinin, 1:500 (Abcam, ab68167); anti-Connexin-43, 1:500 (Sigma, C6219). Secondary antibodies and relevant dilutions were as follows: AlexaFluor 488 anti-mouse, 1:500 (Jackson Immuno, # 715-545-150), AlexaFluor 647 anti-mouse, 1:500 (Jackson Immuno, # 715-605-150), AlexaFluor 488 anti-rabbit, 1:500 (Jackson Immuno, # 715-545-152), AlexaFluor 647 anti-rabbit, 1:500 (Jackson Immuno, # 715-605-152). The quantification of the images was performed with CellProfiler(Stirling et al., 2021 (link)).
Wang H., Keepers B., Qian Y., Xie Y., Colon M., Liu J, & Qian L. (2022). Cross-lineage Potential of Ascl1 Uncovered by Comparing Diverse Reprogramming Regulatomes. Cell stem cell, 29(10), 1491-1504.e9.
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6

Immunostaining Protocol for Flow Cytometry

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Cells were fixed, permeabilized, and immuno-stained using the Fixation/Permeabilization Kit from BD Biosciences. Cells were incubated in 1X fixation/permeabilization solution for 20 minutes, washed once 1X permeabilization/wash solution, stained with primary antibody diluted in 1X permeabilization/wash solution for 30 minutes, washed once 1X permeabilization/wash solution, stained with primary antibody diluted in 1X permeabilization/wash solution for 30 minutes, and lastly washed twice with 1X permeabilization/wash solution before flow cytometry. Samples were analyzed with an Attune NxT cytometer (Thermo Fisher). Gating in the forward and side scatter channels was performed to gate out debris and doublet cells. Primary antibodies and relevant dilutions were as follows: anti-Tuj1, 1:500 (Abcam, # ab7751); anti-Alb, 1:500 (Abcam, ab207327); anti-cTnT, 1:200 (Thermo Fisher, # MA5-12960); FITC-conjugated anti-cTnT, 1:100 (Miltenyi, # 130-119-575); anti-αActinin, 1:250 (Abcam, ab68167). Secondary antibodies and relevant dilutions were as follows: AlexaFluor 488 anti-mouse, 1:500 (Jackson Immuno, # 715-545-150), AlexaFluor 647 anti-mouse, 1:500 (Jackson Immuno, # 715-605-150), AlexaFluor 488 anti-rabbit, 1:500 (Jackson Immuno, # 715-545-152), AlexaFluor 647 anti-rabbit, 1:500 (Jackson Immuno, # 715-605-152).
Wang H., Keepers B., Qian Y., Xie Y., Colon M., Liu J, & Qian L. (2022). Cross-lineage Potential of Ascl1 Uncovered by Comparing Diverse Reprogramming Regulatomes. Cell stem cell, 29(10), 1491-1504.e9.
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7

Immunostaining of Mouse Spermatocyte Chromosomes

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Surface-spread chromosomes of mouse spermatocytes were prepared from 18 days postpartum male mice using the drying-down technique as previously described33 (link). Immuno-staining was performed as previously described33 (link) using the following primary and secondary antibodies: goat anti-SYCP3 (Santa Cruz Biotechnology, sc-20845; 1:200), rabbit anti-MSH4 (a gift from Dr. Paula Cohen; 1:200), mouse monoclonal anti-MLH1 (4C9C7) (Cell Signaling Technology, 3515S; 1:30), AMCA anti-goat (Jackson ImmunoResearch, 705–155-147; 1:50), Cy3 anti-rabbit (Jackson ImmunoResearch, 711–165-152; 1:100), and AlexaFluor647 anti-mouse (Jackson ImmunoResearch, 715–605-151; 1:100).
Kulkarni D.S., Owens S.N., Honda M., Ito M., Yang Y., Corrigan M.W., Chen L., Quan A.L, & Hunter N. (2020). PCNA activates the MutLγ endonuclease to promote meiotic crossing over. Nature, 586(7830), 623-627.
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8

Cryosectioning and Immunofluorescence of Decalcified Cochleae

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Decalcified cochleae were rinsed three times (30 minutes per rinse) in phosphate buffer saline (PBS). The cochleae were then immersed in 30% sucrose overnight for cryoprotection and embedded in OCT compound for cryo-sectioning at −20 °C. Cryosections with a thickness of 10 µm were washed three times before blocking for one hour at room temperature in blocking solution containing 10% goat serum in PBST (0.1% Triton X-100 in PBS). They were then incubated overnight at 4 °C in mouse anti-Tuj1 (Covance) and rabbit anti-peripherin (Millipore), both in an antibody solution of PBS +3% goat serum +0.1% Triton X-100. Sections were washed three times with PBST and then incubated for 2 hours at room temperature in Alexa Fluor 647 anti-mouse, Cy3 goat anti-rabbit, and FITC goat anti-rabbit (Jackson ImmunoResearch Laboratories, INC.). Specimens were washed three times, 5 minutes each, in PBS before mounting with fluorescent mounting medium (Electron Microscopy Sciences).
Lee J.H., Park S., Perez-Flores M.C., Wang W., Kim H.J., Izu L., Gratton M.A., Chiamvimonvat N, & Yamoah E.N. (2019). Early functional alterations in membrane properties and neuronal degeneration are hallmarks of progressive hearing loss in NOD mice. Scientific Reports, 9, 12128.
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9

Immunofluorescence Staining of Neural Proteins

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Tissue sections were permeabilized with PBS/0.3% Tween-20 (PBST) for ~ 30 min then blocked with 5% normal donkey or normal horse serum in 0.01% PBST at room temperature for ~ 1 h. Primary antibodies were diluted in 0.01% PBST and 5% serum then incubated overnight at 4 °C with gentle shaking. Primary antibodies used for this study were; rabbit anti-TARDBP (Proteintech 10,782–2-AP), mouse anti-NEUN (Millipore Sigma MAB377), and DAPI (4′,6-diamidino-2-phenylindole)(Sigma Aldrich 10236276001). After rinsing in PBST 3X, secondary antibodies diluted in 0.01% PBST and 5% serum were added and incubated at room temperature for ~ 1 h with gentle shaking. Secondary antibodies included Alexa Fluor 555 anti-rabbit (Abcam ab150106) and Alexa Fluor 647 anti-mouse (Jackson ImmunoResearch 715-605-151). DAPI was used for nucleus staining. Imaging was performed using a Zeiss LSM 800 laser scanning confocal microscope (LSCM) with Airyscan.
Bjorklund G.R., Wong J., Brafman D., Bowser R, & Stabenfeldt S.E. (2023). Traumatic brain injury induces TDP-43 mislocalization and neurodegenerative effects in tissue distal to the primary injury site in a non-transgenic mouse. Acta Neuropathologica Communications, 11, 137.
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10

Single-Molecule Localization Microscopy Sample Preparation

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For SMLM, coverslips (Precision coverslips, 24 mm, Carl Roth) were cleaned by a 10-min wash in ethanol, rinsed in milli-Q water, and plasma cleaned (PlasmaPrep2, GaLa Instrumente). Clean slides were incubated with 0.01% poly-l-lysine (30,000–70,000 D, Sigma-Aldrich) for 10 min. After incubation, coverslips were rinsed in milli-Q water and dried at room temperature. Poly-l-lysine–coated coverslips were stored at 4°C until use.
Immunofluorescence samples for SMLM imaging were prepared as described above with a few modifications. Worms were immobilized in 0.02% of tetramisole during dissection and fixed with 1% PFA on poly-l-lysine–coated coverslips. Primary antibodies used for SMLM were: anti-HA (1:200, mouse monoclonal 2-2.2.14, Thermo Fisher Scientific), anti-HIM-3 (1:200 or 1:100, rabbit polyclonal 53470002, Novus Biologicals). Secondary antibodies were Alexa Fluor 647 anti-mouse (1:1,000 or 1:500, donkey polyclonal, Jackson Immunoresearch) and CF680 anti-rabbit (1:1,000 or 1:500, goat F(ab)′ fragment, Sigma-Aldrich). The samples were mounted in a custom sample holder and imaged in blinking buffer (50 mM Tris HCl, pH 8, 10 mM NaCl, 10% (wt/vol) d-glucose, 500 µg/ml glucose oxidase, 40 µg/ml catalase, and 35 mM mercaptoethylamine).
Hurlock M.E., Čavka I., Kursel L.E., Haversat J., Wooten M., Nizami Z., Turniansky R., Hoess P., Ries J., Gall J.G., Rog O., Köhler S, & Kim Y. (2020). Identification of novel synaptonemal complex components in C. elegans. The Journal of Cell Biology, 219(5), e201910043.
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