Deidentified, formalin-fixed, paraffin-embedded tissue was procured from four cases of prostate cancer and 10 cases of PDAC through the Cooperative Human Tissue Network with approval from the Vanderbilt Institutional Review Board. 5-μm sections were processed as described (Shi et al., 2014 (link)) and labeled with antibodies to Fn for colorimetric analysis. Detection was performed using Vectastain Elite ABC kit (Vector Laboratories) after a reaction with 3,3′-diaminobenzidine (Vector Laboratories). For fluorescent analysis, antibodies to smooth muscle α-actin (αSMA) were detected using the Vectastain Elite ABC kit, followed by tyramide signal amplification with the TSA Plus Cyanine 3 kit (PerkinElmer), followed by heat inactivation. Binding of anti–Fn antibodies was detected with Cy2–anti-rabbit antibodies (Jackson ImmunoResearch Laboratories, Inc.). Slides were counterstained with hematoxylin (Sigma Aldrich) or Toto3 (Thermo Fisher Scientific) and DAPI (Sigma-Aldrich). Colorimetric images were obtained on an Axioskop 40 microscope (ZEISS), and fluorescent images were captured on the Quorum WaveFX spinning disk confocal system with Nikon Eclipse Ti microscope as described in Microscopy.
Vectastain elite abc kit
Manufactured by PerkinElmer
The Vectastain Elite ABC kit is a reagent system used in immunohistochemistry and immunocytochemistry applications. It facilitates the detection and visualization of target antigens in biological samples. The kit contains the necessary components to perform the avidin-biotin complex (ABC) method of signal amplification.
Automatically generated - may contain errors
Lab products found in correlation
Tsa plus cyanine3 kit, by PerkinElmer (1 mentions)
Axioskop 40 microscope, by Zeiss (1 mentions)
Hematoxylin, by Merck Group (1 mentions)
Toto 3, by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Vectastain elite abc kit, by Vector Laboratories (1 mentions)
3 3 diaminobenzidine (dab), by Vector Laboratories (1 mentions)
Nel753001kt, by PerkinElmer (1 mentions)
Biotin conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)
2 protocols using vectastain elite abc kit
1
Immunohistochemical Analysis of Prostate and Pancreatic Tumors
2
In situ mRNA detection in Drosophila embryos
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Dechorionated Drosophila embryos were fixed in paraformaldehyde-heptane solution for 30 min, followed by devitellinization using methanol. Freshly fixed embryos were then labeled with digoxigenin (DIG)-labeled antisense probes following the standard procedure described in Knirr et al., 1999 (link). After rehydration and post-fixation in 10% formaldehyde, embryos were pre-hybridized for 1 hr at 55 °C. mRNA probes were added to the embryos which were then incubated for 16 hr at 55 °C. DIG present on the synthesized mRNA probes was subsequently labeled with anti-DIG primary antibody (Enzo; ENZ-ABS302-0001; 1:1,000) and biotin-conjugated secondary antibody (Invitrogen; 1:500). To detect fluorescent signals, embryos were incubated with AB solution (PK-6100, Vectastain Elite ABC kit) followed by the tyramide signal amplification reaction (Cy3 fluorescent dye in amplification diluent; 1:50) (Perkin Elmer Life Sciences, NEL753001kt). To mark the SG and the apical membrane, the embryos were immunolabeled with antibodies for CrebA (SG nuclei) and Crb (apical membrane).
Primer sequences used for mRNA probe synthesis in this study are listed below.
Primer sequences used for mRNA probe synthesis in this study are listed below.
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