Slides with 4 μm thick sections of mouse or human pancreatic tissues embedded in paraffin were deparaffinised. The sections were unmasked using unmasking solution (Vector H 3300), saturated with Ab diluent (Dako) for 30 min and then incubated with primary Abs (anti-βig-h3, Sigma; anti-caspase-3, Cell Signaling; and CK19 Troma III, DSHB) that were diluted in Ab diluent overnight at 4°C. The sections were washed and then incubated with goat antirat biotinylated secondary Abs (BD Biosciences; 1:200) for 1 hour at RT. The remaining steps were performed using Vectastain ABC kits (Vector Labs). The slides were counterstained with hematoxylin. Alternatively, immunofluorescence was performed using deparaffinised and unmasked sections, which were incubated in anti-βig-h3, anti-PDGRFα, anti-EPCAM, anti-GrzB and anti-Casp3 primary Abs overnight at 4°C and then with specific anti-Fab′2-Alexa 647 and anti-Fab ′ 2-Alexa 555 (Molecular Probes) secondary Abs. Finally, the sections were mounted in Vectashield mounting medium with DAPI.
Vector h 3300
Manufactured by Vector Laboratories
Sourced in United Kingdom
The Vector H-3300 is a high-performance laboratory equipment designed for accurate and reliable measurements. It is a versatile instrument that can be used for a wide range of applications. The core function of the Vector H-3300 is to provide precise and consistent data collection and analysis.
Automatically generated - may contain errors
Lab products found in correlation
Vector sk 4100, by Vector Laboratories (2 mentions)
Ab diluent, by Agilent Technologies (1 mentions)
Anti fab 2 alexa 647, by Thermo Fisher Scientific (1 mentions)
Vectastain abc kit, by Vector Laboratories (1 mentions)
Anti caspase 3, by Cell Signaling Technology (1 mentions)
Vectashield with dapi, by Vector Laboratories (1 mentions)
Anti erk1, by Thermo Fisher Scientific (1 mentions)
C2 system, by Nikon (1 mentions)
Anti erk2, by Cell Signaling Technology (1 mentions)
Non specific blocking reagent, by Agilent Technologies (1 mentions)
Anti p erk1 2, by Cell Signaling Technology (1 mentions)
Anti ctbp2, by BD (1 mentions)
Vectastain universal elite abc kit, by Vector Laboratories (1 mentions)
Hpa005692, by Merck Group (1 mentions)
Vector ba 1000, by Vector Laboratories (1 mentions)
Vector pk 4001, by Vector Laboratories (1 mentions)
6 protocols using vector h 3300
1
Immunohistochemical Analysis of Mouse and Human Pancreatic Tissues
2
Tissue Microarray of High-grade Serous Ovarian Tumors
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FFPE blocks of high-grade serous ovarian tumors were collected under the approval of the Institutional ethical committee from the archive of the Department of Pathology at the Medical Center. The tumors were removed from patients undergoing either primary cytoreductive or interval debulking surgery at the Medical Center since 2011. To construct a tissue microarray,49 (link) representative cores of tissue (1.5 mm in diameter) were taken from each case with a total of 46 different cases. Tumor fragments were rinsed with ice cold PBS. Specimens were fixed in 10% formalin overnight at room temperature and paraffin embedded. Sections of 5 µm were cut using a microtome, deparaffinized and rehydrated through a graded ethanol series, unmasked (Vector H-3300), blocked 2 h in PBS/0.2% Tween20/0.2% gelatin (Sigma G-1890), and stained with hematoxylin and eosin (H&E) for histology examination.
3
Immunolabeling of Cellular Proteins
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Paraffin-embedded sections were deparaffinized and immersed in unmasking solution (Vector H3300; Vector Laboratories) for antigenic retrieval and heated in an autoclave (121 °C) for 5 min. Sections were then incubated with a non-specific blocking reagent (Dako) for 1 h to block any non-specific reactions, followed by overnight incubation at 4 °C with one of the following primary antibodies: anti-ERK1 (Invitrogen, 13–8600), anti-ERK2 (Cell Signaling, #9108), anti-p-ERK1/2 (Cell Signaling, #9101), anti-myosin VIIa (Proteus Biosciences, 25–6790), or anti-CtBP2 (BD Transductions, 612044) diluted in an antibody diluent (DAKO). Sections were washed thrice in PBS and incubated with a corresponding secondary antibody (Alexa Fluor 488 or 546, IgG, Invitrogen) diluted in an antibody diluent (DAKO). After rinsing in PBS, NSE was mounted onto slides with an antifade mounting medium (VECTASHIELD with DAPI; Vector Laboratories, Burlingame, CA, USA). DAPI labeling was then used to identify condensed HC nuclei. Images of immunolabeled specimens were obtained by confocal fluorescence microscopy using a Nikon C2 system (Nikon, Tokyo, Japan).
4
Immunohistochemical Staining of Paraffin-Embedded Tissues
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Paraffin-embedded tissue array sections were deparaffinized and treated with antigen unmasking solution (Vector H-3300) according to the manufacturer's instructions. Peroxidase block was performed by incubating the slides for 20–30 min in 0.3% H2O2. After a rinse with H2O, slides were first blocked with diluted normal horse serum (R.T.U. Vectastain Elite ABC kit universal) for 20 min and subsequently with the Biotin/Avidin blocking system by Vector. Sections were then stained with the primary antibody (rabbit anti N-term p32 5μg/ml, rabbit anti Myc (1:200) in normal horse serum for 1 h at RT or overnight at 4°C. Antibody binding was detected after a 30-min incubation with biotinylated “universal” secondary antibody and VECTASTAIN RT Elite ABC reagent. Peroxidase substrate solution (DAB substrate kit) was incubated until desired stain intensity developed. Slides were finally counterstained with Hematoxylin (Vector H-3401). The slides were scanned on a Scanscope CM-1 scanner and subsequently processed for staining quantification using ImageScope software (Aperio Technology).
5
Liver Transferrin Immunohistochemistry
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For immunohistochemistry, 3–4-μm-thick liver sections were deparaffinized and rehydrated, and a citrate-based antigen retrieval was performed (Vector H-3300, Vector Laboratories, Petersborough, UK) for 30 min, as recommended by the supplier. The immunodetection was carried out with anti-transferrin as a primary (11,000 dilution; Sigma; HPA005692, Stockholm, Sweden) and a biotin-conjugated anti-rabbit secondary antibody (Vector BA1000). The signal was visualized with peroxidase-labeled streptavidin and the 3,3′-diaminobenzidine as a substrate (Vector SK4100).
6
Immunohistochemical Analysis of HSL
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