Cloned embryos were washed in PBS containing 0.1% polyvinyl alcohol (PVA) and then fixed in 4% (w/v) paraformaldehyde at room temperature for 30 min. The oocytes were then washed in PBS/PVA and incubated overnight in PBS containing 1% BSA and 0.1% Triton X-100 at 4°C. Following this, the oocytes were washed three times in PBS-0.1% BSA and incubated with a 1:200 dilution of H3K9me3 antibody (07–442, Millipore) at the 1- and 2-cell stages and purified mouse anti-OCT-3/4 (611203, BD Bioscience) at the blastocyst stage for 2 h at room temperature. The cloned embryos were then washed three times in PBS-0.1% BSA and incubated with a 1:200 dilution of goat anti-mouse antibody (ab150113, Abcam) for 1 h at room temperature. Following this, they were washed in PBS-0.1% BSA three additional times. The DNA was visualized by staining the oocytes with 4′,6-diamidino-2-phenylindole (DAPI; D5942, Sigma-Aldrich). The embryos were mounted on glass slides with a drop of fluorescent mounting medium and then observed using fluorescence confocal microscopy (Zeiss LSM880; Zeiss, Jena, Germany).
Purified mouse anti oct 3 4
Manufactured by BD
Sourced in United States
Purified mouse anti-OCT-3/4 is a laboratory reagent used for the detection and identification of the OCT-3/4 protein, which is a transcription factor involved in the regulation of embryonic stem cell pluripotency. This product provides a specific and reliable tool for researchers studying stem cell biology and development.
Automatically generated - may contain errors
Lab products found in correlation
Ab150113, by Abcam (1 mentions)
Lsm 880, by Zeiss (1 mentions)
H3k9me3 antibody, by Merck Group (1 mentions)
D5942, by Merck Group (1 mentions)
Alexa fluor 594 goat anti mouse igg h l, by Thermo Fisher Scientific (1 mentions)
Normal goat serum (ngs), by Vector Laboratories (1 mentions)
Mounting medium with dapi, by ZSGB-BIO (1 mentions)
Alexa fluor 488 goat anti mouse igg h l, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 594 goat anti rabbit igg h l, by Thermo Fisher Scientific (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Dm3000 microscope, by Leica (1 mentions)
2 protocols using purified mouse anti oct 3 4
1
Immunofluorescence Analysis of Cloned Embryos
2
Immunofluorescence Analysis of Neural Crest Markers
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Cells cultured on Matrigel-precoated glass coverslips were fixed, permeabilized followed by blocking with PBS containing 5% normal goat serum (Vector Laboratories, Burlingame, CA) and 2% BSA (Sigma) at room temperature. Incubation was performed overnight at 4°C with primary antibodies, including p75NTR (D4B3) Rabbit mAb (CST, 8238, Danvers, MA), SOX9 Rabbit mAb (CST, 82630), SOX10 Rabbit mAb (Abcam, ab155279, Cambridge, UK), AP2α+β Mouse mAb (Abcam, A6/2/2 ), FOXD3 polyAb (Biolegend, 631702, San Diego, CA), CD57 Mouse mAb (Santa Cruz, sc-81633, Dallas, Texas), Pax-6 (AD2.35) (Santa Cruz, sc-53108), Purified Mouse Anti-Oct-3/4 (BD Bioscience, 611203), Sox2 (D9B8N) Rabbit mAb (CST, 23064S), and Peripherin Rabbit polyAb (Abcam, ab4666). The primary antibody incubations were followed with appropriate secondary antibodies: Alexa Fluor 594 goat anti-rabbit IgG (H+L) (Invitrogen, A-11037), Goat Alexa Fluor 488 goat anti-mouse IgG (H+L) (Invitrogen, A-11029), Alexa Fluor 594 goat anti-mouse IgG (H+L) (Invitrogen, A-11005), Alexa Fluor 488 goat anti-mouse IgG (H+L) (Invitrogen, A-11001). The slides were mounted with mounting medium with DAPI (ZSGBBio, Beijing, China) to localize the nucleus and then imaged with a Leica DM3000 microscope (Leica Microsystems Gmbh, Wetzlar, Germany).
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