Tissue sections were blocked for 1.5 h in 5% milk or 3% BSA dissolved in 0.1% Triton-X/PBS (PBSTx). Wild-type sections were used to examine protein expression of TFAP2A (3B5-supernatant, Santa Cruz Biotechnology; 1:25 dilution) and TFAP2B (2509S, Cell Signaling Technology; 1:25 dilution). E11.5 control and Tfap2NCKO midface sections were stained with phosphorylated Histone H3 (9701S, Cell Signaling Technology; 1:400 dilution) to assess cell proliferation. After washing with PBSTx, samples were incubated with Alexa Fluor 488/568 secondary antibodies (A-11017, A-11070 and A-21069, Invitrogen; 1:500 dilution) for 1.5 h. We used DAPI to counterstain nuclei. Stains were imaged via Zen software on a Zeiss 700 LSM confocal microscope. Images were exported as Z-stacked, maximum intensity projection TIFF images.
3b5 supernatant
Manufactured by Santa Cruz Biotechnology
3B5-supernatant is a cell culture supernatant product from Santa Cruz Biotechnology. It is derived from the 3B5 hybridoma cell line. The product is intended for research use only.
Automatically generated - may contain errors
2 protocols using 3b5 supernatant
1
Immunofluorescence Staining of Midface Tissue
2
Immunofluorescent Staining of TFAP2A and TFAP2B
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Sectioned tissue was blocked for 1.5 hours in 5% milk or 3% BSA dissolved in 0.1% Triton-X/PBS (PBSTx). Wild-type sections were used to examine protein expression of TFAP2A (3B5-supernatant, Santa Cruz Biotechnology; 1:25 dilution) and TFAP2B (2509S, Cell Signaling Technology; 1:25 dilution). After washing with PBSTx, samples were incubated with Alexa Fluor 488/568 secondary antibodies (Invitrogen; 1:500 dilution) for 1.5 hours. We used DAPI to counterstain nuclei. Stains were imaged via Zen software on a Zeiss 700 LSM confocal microscope. Images were exported as Z-stacked, maximum intensity projection TIFF images.
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