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2 protocols using ab70383

1

Immunofluorescence Localization of Cell Cycle Proteins

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Primary antibodies: rabbit‐anti‐MAD2 (1:200; ab70385, abcam; or 1:500 ab70383, abcam); sheep‐anti‐tryptophan hydroxylase/TPH (1:150; ab32821, abcam); rabbit‐anti‐BubR1 (1:100; A300‐386A, Bethyl Laboratories Inc.); rabbit‐anti‐P31comet (murine) (1:100; provided by Dr. Hongtao Yu; UT Southwestern, TX, USA); mouse‐anti‐SERT (H‐45) (1:20; raised and provided by Dr. Egon Ogris, Max Perutz Labs, Vienna, Austria (Montgomery et al, 2014 (link))); rabbit‐anti‐SEC24C (1:100; provided by Dr. Randy Schekman, UC Berkeley, CA, USA); rabbit‐anti‐GM130 (1:200; ab52649, abcam); rabbit‐anti‐β‐COP (1:100; PA1‐061, Invitrogen); rabbit‐anti‐TGN38 (1:100; OST00228G‐500UG, Osenses) rabbit‐anti‐Rab7 (1:100; 9367S, Cell Signaling); rabbit‐anti‐Rab5 (1:100; 3547S; Cell Signaling); rabbit‐anti‐Rab11A (1:100; 71‐5300; Invitrogen); rabbit‐anti‐Rab4 (1:100; PA3912, Invitrogen); and rabbit‐anti‐LAMP1 (1:100, L1418; Sigma‐Aldrich). Secondary antibodies: Alexa Fluor 488 donkey‐anti‐rabbit IgG (1:500; A21206, Invitrogen); Alexa Fluor 555 donkey‐anti‐sheep IgG (1:500; A21436, Invitrogen); Alexa Fluor 555 donkey‐anti‐mouse (1:500; A31570, Invitrogen); and Alexa Fluor 647 goat‐anti‐rabbit (1:500; A32733, Invitrogen).
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2

Immunofluorescence Staining of Mitotic Markers

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The following primary antibodies were used: anti‐γ‐H2AX (Abcam, ab22551, 1:250), anti‐MDC1 (Abcam, ab241048, 1:100), anti‐acetylated‐α‐tubulin (Sigma‐Aldrich, T7451, 1:500; Abcam, ab179484, 1:500), anti‐Mad2 (Abcam, ab70383, 1:100), anti‐BubR1 (Abcam, ab28193, 1:100), anti‐centromere (ACA; Antibodies Incorporated, 15–234, 1:100), anti‐SMC3 (Abcam, ab128919, 1:100), anti‐SMC4 (Novus Biologicals, NBP1‐86635, 1:100) and anti‐CENP‐A (Cell Signaling, #2048, 1:250). Alexa Fluor 488‐conjugated anti‐mouse (Jackson ImmunoResearch, 115‐545‐144, 1:500), Alexa Fluor 488‐conjugated anti‐rabbit (Jackson ImmunoResearch, 115‐545‐144 1:500), Alexa Fluor 594‐conjugated anti‐rabbit (Jackson ImmunoResearch, 111‐585‐144, 1:500) and Rhodamine (TRITC) ‐conjugated anti‐human (Jackson ImmunoResearch, 109‐025‐088, 1:100) were used as secondary antibodies.
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