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Alexa fluor conjugated series

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The Alexa Fluor-conjugated series are a family of fluorescent dyes developed for use in various biological applications. These dyes are characterized by their bright fluorescence, photostability, and ability to be conjugated to a wide range of biomolecules. The Alexa Fluor dyes are available in multiple emission wavelengths, allowing for flexibility in experimental design and multi-color analysis.

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Lab products found in correlation

H3663, by Santa Cruz Biotechnology (2 mentions) Sc 56, by Abcam (2 mentions) Anti mcherry, by Thermo Fisher Scientific (2 mentions) M11217, by Merck Group (2 mentions) Anti h4k20me2 3, by Abcam (2 mentions) Lsm 700 multiphoton confocal microscope, by Adobe (2 mentions) Anti h3k27me3, by Merck Group (2 mentions) Lsm 510 meta, by Zeiss (2 mentions) Rabbit anti vasa, by Santa Cruz Biotechnology (1 mentions) Lysotracker, by Thermo Fisher Scientific (1 mentions) Alkaline phosphatase detection kit, by Merck Group (1 mentions)

Close spelling variants of this product

Alexa Fluor 488- and 555-conjugated antibody series against mouse, Alexa Fluor series Alexa series Alexa Fluors

4 protocols using alexa fluor conjugated series

1

Immunofluorescence Staining of Cellular Markers

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Immunofluorescence staining was performed using standard procedures7 (link),71 . Primary antibodies used were mouse anti-Fas III (1:200, DSHB, 7G10), anti-HA (1:200; Sigma-Aldrich H3663), anti-PCNA (1:200; Santa Cruz sc-56), anti-GFP (1:1,000; Abcam ab 13970), anti-mKO (1:200; MBL PM051M), anti-mCherry (1:1000; Invitrogen M11217), anti-H3K27me3 (1:200; Millipore 07–449), anti-H4K20me2/3 (1:400; Abcam ab7817), anti-ssDNA (1:100, DSHB) and anti-BrdU (1:200; Abcam ab6326). BrdU analogue was Invitrogen B23151 5-bromo-2´-deoxyuridine (BrdU). Secondary antibodies were the Alexa Fluor-conjugated series (1:1000; Molecular Probes). Confocal images for immunostained fixed sample were taken using Zeiss LSM 700 Multiphoton confocal microscope with 63x or 100x oil immersion objectives and processed using Adobe Photoshop software.
Wooten M., Snedeker J., Nizami Z.F., Yang X., Ranjan R., Urban E., Kim J.M., Gall J., Xiao J, & Chen X. (2019). Asymmetric histone inheritance via strand-specific incorporation and biased replication fork movement. Nature structural & molecular biology, 26(8), 732-743.
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2

Immunofluorescence Staining of Cellular Markers

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Immunofluorescence staining was performed using standard procedures7 (link),71 . Primary antibodies used were mouse anti-Fas III (1:200, DSHB, 7G10), anti-HA (1:200; Sigma-Aldrich H3663), anti-PCNA (1:200; Santa Cruz sc-56), anti-GFP (1:1,000; Abcam ab 13970), anti-mKO (1:200; MBL PM051M), anti-mCherry (1:1000; Invitrogen M11217), anti-H3K27me3 (1:200; Millipore 07–449), anti-H4K20me2/3 (1:400; Abcam ab7817), anti-ssDNA (1:100, DSHB) and anti-BrdU (1:200; Abcam ab6326). BrdU analogue was Invitrogen B23151 5-bromo-2´-deoxyuridine (BrdU). Secondary antibodies were the Alexa Fluor-conjugated series (1:1000; Molecular Probes). Confocal images for immunostained fixed sample were taken using Zeiss LSM 700 Multiphoton confocal microscope with 63x or 100x oil immersion objectives and processed using Adobe Photoshop software.
Wooten M., Snedeker J., Nizami Z.F., Yang X., Ranjan R., Urban E., Kim J.M., Gall J., Xiao J, & Chen X. (2019). Asymmetric histone inheritance via strand-specific incorporation and biased replication fork movement. Nature structural & molecular biology, 26(8), 732-743.
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3

Immunofluorescence Staining and Microscopy Protocol

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Immunofluorescence staining was performed using standard procedures (Hime et al., 1996 (link); Tran et al., 2012 (link)). Primary antibodies were mouse anti-α spectrin (1:50, DSHB 3A9), mouse anti-Fas III (1:50, DSHB, 7G10), mouse anti-Armadillo (1:100; DSHB, N2 7A1 clone), rabbit anti-H3T3P (1:200, Millipore 05-746R), mouse anti-H3S10P (1:200; Millipore, #05-806), chicken anti-CID (1:100; gift from Dr. Sylvia Erhardt, University of Heidelberg, Germany), and rabbit anti-Vasa (1:200; Santa Cruz SC-30210). Secondary antibodies were the Alexa Fluor-conjugated series (1:200; Molecular Probes). Lysotracker (Invitrogen L7528) is applied according to manufacturer recommendation. Images were taken using the Zeiss LSM 510 META or Zeiss LSM 700 Multiphoton confocal microscope with a 40x or 63 x oil immersion objectives and processed using Adobe Photoshop software.
Xie J., Wooten M., Tran V., Chen B.C., Pozmanter C., Simbolon C., Betzig E, & Chen X. (2015). Histone H3 Threonine Phosphorylation Regulates Asymmetric Histone Inheritance in the Drosophila Male Germline. Cell, 163(4), 920-933.
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4

Immunofluorescence Staining of Mouse ESCs

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Immunofluorescence staining of mESCs was performed using standard procedures (Hime et al., 1996 (link)). Primary antibodies were rabbit anti-APC (1:150, Santa Cruz sc-7930), goat anti-Claudin 6 (1:150, Santa Cruz sc-17669), rabbit anti-Sox2 (1:200, Active Motif 39823), rat anti-Nanog (1:200, Active Motif 61627), rabbit anti-Rex1 (1:200, Thermo Fisher# PA5-27567), rabbit anti-Oct4 (1:200, Abcam# ab181557), mouse anti-SSEA-1 (1:50, Thermo Fisher# MC-480). Secondary antibodies were the Alexa Fluor-conjugated series (1:600, Molecular Probes). ESCs and EpiSCs were also stained with an Alkaline Phosphatase Detection Kit (SCR004, Millipore) following the manufacturer’s instructions. Culture the cells for 5 days prior to analyzing ALP activity. Then fix the cells with 4% paraformaldehyde (PFA) in PBS for 1-2 mins. Aspirate the fixative and rinse with 1× Rinse buffer for twice. Prepare the Alkaline Phosphatase staining solution by mixing Fast Red Violet (FRV) with Napthol AS-BI phosphate solution and water in a 2:1:1 ratio. Incubate the cells in dark at room temperature for 30 mins. Aspirate the staining solution and rinse with 1× Rinse buffer for twice. Cover the cells with PBS and then image the colonies with ALP activity. Related to validation Figure A-B and expression Figure E in Mendeley data.
Ma B., Trieu T.J., Cheng J., Zhou S., Tang Q., Xie J., Liu J.L., Zhao K., Habib S.J, & Chen X. (2020). Differential Histone Distribution Patterns in Induced Asymmetrically Dividing Mouse Embryonic Stem Cells. Cell reports, 32(6), 108003.
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