Immunofluorescence Staining of Histone Marks

Primary antibodies and their dilutions were as follows: 1:30,000 of 2.45 mg/mL mouse anti-H3K27me3 (Kimura mAb 1E7 clone CMA323, Wako cat#309-95259), 1:3000 original stock solution of rabbit anti-H3K27me3 (C36B11 Cell Signaling MAb#9733 lot#C36B11), 1:4000 of 0.4 mg/mL mouse anti-GFP (Roche cat#11 814 460 001 lot#14158300), 1:4000 rabbit anti-UNC-64 serum²², 1:50,000 rabbit anti-PGL-1 serum²³, 1:500 guinea pig anti-HTP-3 serum^{24 (link)}. Secondary antibodies conjugated to Alexa Fluor: 488 goat anti-mouse (Life cat#A11001), 488 goat anti-rabbit (Molecular Probes cat#A-11008), 594 goat anti-rabbit (Life cat#A11012), and 594 goat anti-guinea pig (Molecular Probes cat#A11076) were used at 1:300 with 0.05 μg/mL 4′,6-diamidino-2-phenylindole (DAPI).

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Kaneshiro K.R., Rechtsteiner A, & Strome S. (2019). Sperm-inherited H3K27me3 impacts offspring transcription and development in C. elegans. Nature Communications, 10, 1271.

Publication 2019

488 goat anti-rabbit

Antibodies Clone Dapi Dilutions Goat Guinea pig Molecular probes Mouse Rabbit

Corresponding Organization :

Other organizations : University of California, Santa Cruz

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Variable analysis

independent variables

Concentrations of primary antibodies

dependent variables

Not explicitly mentioned

control variables

Concentrations of secondary antibodies (1:300)
Concentration of DAPI (0.05 μg/mL)

controls

Positive controls: Not explicitly mentioned
Negative controls: Not explicitly mentioned

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