Fixed pESCs were incubated with anti-vimentin (1:200, ab92547) and anti-CK7 (1:200, ab181598) (both from Abcam, Cambridge, MA, USA), and fixed ESCs from different treatments were incubated with anti-PRL (1:150, ab64377; Abcam) primary antibody overnight at 4°C separately, washed with PBS and incubated with FITC-conjugated secondary antibody (1:200, ab6717; Abcam). After washing with PBS, cells were mounted with anti-fading mounting medium with DAPI and observed with a fluorescence microscope (magnification, ×40). Images were captured with a charge coupled device camera.
Ab64377
Manufactured by Abcam
Sourced in United States
Ab64377 is a lab equipment product offered by Abcam. It serves as a core function in various laboratory applications.
Automatically generated - may contain errors
3 protocols using ab64377
1
Immunofluorescence Staining of Stem Cells
2
Immunohistochemical Analysis of Pituitary Prolactin
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Serial 4-μm sections were prepared from formalin-fixed, paraffin-embedded tissue specimens obtained from the pituitary and choroid plexus. Rabbit polyclonal antibody to PRL (ab64377; Abcam) was used at empirically determined dilutions [24 (link)]. Immunoreactivity was achieved by the polymer method using Dako Envison+ Dual Link System-HRP (K4063; Dako, California, USA) and Dako Liquid DAB+ Substrate Chromogen System (K3468; Dako) according to the manufacturer’s instructions. The percent positivity in the pituitary was estimated as the percentage of positive PRL cells, which was calculated as follows: number of positive PRL cells / total number of anterior pituitary cells × 100 [25 (link)].
3
Pituitary Prolactin Immunohistochemistry
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Serial 4-µm sections were prepared from formalin-fixed, paraffin-embedded tissue specimens obtained from the pituitary. Rabbit polyclonal antibody to PRL (ab64377; Abcam) was used at empirically determined dilutions. Immunoreactivity was achieved by the polymer method using Dako EnVision+ Dual Link System-HRP (K4063; Dako, CA, USA) and Dako Liquid DAB+ Substrate Chromogen System (K3468; Dako) according to the instructions from the manufacturer.
Percent positivity in the pituitary was estimated as the percentage of positive PRL cells, which was calculated as follows: number of positive PRL cells / total number of anterior pituitary cells × 100.
Percent positivity in the pituitary was estimated as the percentage of positive PRL cells, which was calculated as follows: number of positive PRL cells / total number of anterior pituitary cells × 100.
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