About 1 × 106 mouse ESCs were injected subcutaneously into SCID mice (Taconic) as previously described (12 (link)). Mice were euthanized after 6 weeks, tumors were harvested and fixed in formalin for 2 days, embedded in paraffin blocks and sectioned. Sections were stained with hematoxylin and eosin for histological analysis following standard procedures. Immunohistochemical staining for Plf (anti-Proliferin 1:100, R&D AF1623) was also performed using standard protocols.
Af1623
Manufactured by R&D Systems
Sourced in United States
AF1623 is a recombinant human protein that functions as a growth factor. It belongs to the Fibroblast Growth Factor family.
Automatically generated - may contain errors
Lab products found in correlation
Scid mice, by Taconic Biosciences (1 mentions)
Vectashield with dapi, by Vector Laboratories (1 mentions)
Af305 na, by R&D Systems (1 mentions)
Af4666, by R&D Systems (1 mentions)
Goat anti endomucin, by R&D Systems (1 mentions)
Alexa fluor 488 donkey anti rabbit igg h l, by Jackson ImmunoResearch (1 mentions)
3 protocols using af1623
1
Subcutaneous Teratoma Formation Assay
2
Placental Proliferation Analysis
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Placentas were harvested at 18.5 dpc, and IF assay was performed. Goat anti-proliferin (R&D Systems, AF1623, 1:200) was used as the primary antibody. DAPI was used for nuclear staining.
The number of pixels in the whole cell and nucleus were counted using ImageJ. For statistical analysis, Student’s t-test was performed using R ver. 4.2.0.
The number of pixels in the whole cell and nucleus were counted using ImageJ. For statistical analysis, Student’s t-test was performed using R ver. 4.2.0.
3
Immunofluorescence Staining of Murine Placental Tissue
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Placentas at 18.5 dpc were collected and fixed overnight in 4% PFA at 4°C. The tissues were then processed for paraffin embedding. Sections of size 5 μm thickness were sliced,
deparaffinized, and rehydrated. Antigen retrieval was performed by boiling the sections for 20 min in a citrate-based antigen-unmasking solution. Non-specific binding was blocked using 4%
normal horse serum for 30 min.
The following primary antibodies were used: Rabbit anti-Nrk (Merck, HPA017238, 1:100), goat anti-endomucin (R&D Systems, MN, USA, AF4666, 1:200), goat anti-proliferin (R&D Systems,
AF1623, 1:200), and goat anti-IGF1r (R&D Systems, AF305-NA, 1:200), which were incubated overnight at 4°C. The secondary antibodies used were Alexa Fluor® 488 donkey anti-rabbit IgG
H&L (Jackson ImmunoResearch, 711-545-152, 1:500) and Alexa Fluor® 594 donkey anti-goat IgG H&L (Abcam, Cambridge, UK, ab150132, 1:500), which were incubated for 1 h at 20–25°C.
Counterstaining and mounting were performed using VECTASHIELD® with DAPI (Vector Laboratories, H-1200).
deparaffinized, and rehydrated. Antigen retrieval was performed by boiling the sections for 20 min in a citrate-based antigen-unmasking solution. Non-specific binding was blocked using 4%
normal horse serum for 30 min.
The following primary antibodies were used: Rabbit anti-Nrk (Merck, HPA017238, 1:100), goat anti-endomucin (R&D Systems, MN, USA, AF4666, 1:200), goat anti-proliferin (R&D Systems,
AF1623, 1:200), and goat anti-IGF1r (R&D Systems, AF305-NA, 1:200), which were incubated overnight at 4°C. The secondary antibodies used were Alexa Fluor® 488 donkey anti-rabbit IgG
H&L (Jackson ImmunoResearch, 711-545-152, 1:500) and Alexa Fluor® 594 donkey anti-goat IgG H&L (Abcam, Cambridge, UK, ab150132, 1:500), which were incubated for 1 h at 20–25°C.
Counterstaining and mounting were performed using VECTASHIELD® with DAPI (Vector Laboratories, H-1200).
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