Blastocyst cell counting46 (link) was performed by fixing blastocysts in 4% (w/v) paraformaldehyde and incubating them at 4 °C overnight in rabbit anti-Oct4 antibody (1:100; Santa Cruz) or for 1 h at 37 °C in mouse anti-Cdx2 antibody (1:100; BioGenex, USA), followed by 1 h at 37 °C in Alexa 488-conjugated anti-rabbit IgG (Invitrogen) or Alexa 594-conjugated anti-mouse IgG (Invitrogen) respectively. Cells stained with Alexa 488 were scored as Oct4-positive (pluriblasts) and those with Alexa 594, as Cdx2-positive (trophoblasts).
Rabbit anti oct4 antibody
Manufactured by Santa Cruz Biotechnology
Sourced in United States
The Rabbit anti-Oct4 antibody is an immunoaffinity-purified antibody that recognizes the transcription factor Oct4, which is a key regulator of pluripotency in embryonic stem cells. This antibody can be used for the detection of Oct4 in various applications, such as Western blotting, immunohistochemistry, and flow cytometry.
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Lab products found in correlation
Alexa 488 conjugated anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Alexa594 conjugated anti mouse igg, by Thermo Fisher Scientific (1 mentions)
Mouse anti cdx2 antibody, by BioGenex (1 mentions)
Goat anti pdx1, by R&D Systems (1 mentions)
Mouse anti glucagon, by Merck Group (1 mentions)
Goat anti sox17 antibody, by R&D Systems (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Dojindo Laboratories (1 mentions)
Rat anti c peptide, by Developmental Studies Hybridoma Bank (1 mentions)
Prolong gold antifade reagent, by Thermo Fisher Scientific (1 mentions)
Inverted fluorescent microscope, by Keyence (1 mentions)
Mouse anti map 2 antibody, by Santa Cruz Biotechnology (1 mentions)
3 protocols using rabbit anti oct4 antibody
1
Blastocyst Cell Counting by Immunofluorescence
2
Immunofluorescence Staining of Pluripotent and Endocrine Markers
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Cells were fixed and immunostained with a standard protocol (Ninomiya et al. 2014 (link)). Antibodies used were listed as follows: rabbit anti-OCT4 antibody (1/500; Santa Cruz, Dallas, TX; sc-9081), goat anti-SOX17 antibody (1/300; R&D, Minneapolis, MN; AF1924), goat anti-PDX1 (1/300; R&D; AF2419), mouse anti-NGN3 (1/300; Developmental Studies Hybridoma Bank, Iowa City, IA; F25A1B3), mouse anti-GLUCAGON (1/600; Sigma-Aldrich, St. Louis, MO; G2654), rat anti-C-peptide (1/600; Developmental Studies Hybridoma Bank; GN-ID4), anti-rabbit IgG, Alexa Fluor 488 conjugated (1/600; Invitrogen; A21206), anti-goat IgG, Alexa Fluor 594 conjugated (1/600; Invitrogen; A11058), anti-mouse IgG, Alexa Fluor 488 conjugated (1/600; Invitrogen; A21202), anti-rat IgG, Alexa Fluor 594 conjugated (1/600; Invitrogen; A-11007). Nuclei were counterstained with DAPI (1/200; Dojindo, Kumamoto, Japan; 340–07971) before specimens were mounted in Prolong Gold Antifade Reagent (Invitrogen). Specimens were observed with an inverted fluorescent microscope (Keyence, Osaka, Japan).
3
Immunohistochemistry of Stem Cell Markers
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AKP was stained using a specific kit (Cat. No. ZLI9041; Beijing Zhongshan Jinqiao Co., Beijing, China) following the manufacturer protocol. OCT-4 and MAP-2 were immunostained in germline stem cells and colonies using monospecific antibodies.
In brief, cells and colonies were fixed in 4% paraformaldehyde for 15 min, washed twice with 0.1% bovine serum albumin (BSA) in PBS for 5 min, and blocked with 5% BSA in PBS at room temperature for 40 min. Thereafter, specimens were incubated overnight at 4°C with a rabbit anti-OCT-4 antibody (1:500 dilution; Santa Cruz Biotechnology, Dallas, TX, USA) or a mouse anti-MAP-2 antibody (1:500 dilution; Santa Cruz Biotechnology).
After washing, specimens were incubated with fluorescein isothiocyanate-labeled goat anti-rabbit or anti-mouse immunoglobulin G antibodies (Santa Cruz Biotechnology) at room temperature in the dark for 1 h. Following further washing, the specimens were mounted and examined under a fluorescent microscope. Primary and secondary antibodies were prepared in 1% BSA in PBS. PBS served as the negative control.
In brief, cells and colonies were fixed in 4% paraformaldehyde for 15 min, washed twice with 0.1% bovine serum albumin (BSA) in PBS for 5 min, and blocked with 5% BSA in PBS at room temperature for 40 min. Thereafter, specimens were incubated overnight at 4°C with a rabbit anti-OCT-4 antibody (1:500 dilution; Santa Cruz Biotechnology, Dallas, TX, USA) or a mouse anti-MAP-2 antibody (1:500 dilution; Santa Cruz Biotechnology).
After washing, specimens were incubated with fluorescein isothiocyanate-labeled goat anti-rabbit or anti-mouse immunoglobulin G antibodies (Santa Cruz Biotechnology) at room temperature in the dark for 1 h. Following further washing, the specimens were mounted and examined under a fluorescent microscope. Primary and secondary antibodies were prepared in 1% BSA in PBS. PBS served as the negative control.
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