After the blocking procedure with 1.5% donkey serum (#sc-2023, Santa Cruz Biotechnology, Inc.), the slides were incubated overnight at 4˚C with the goat anti-XBP1 antibody (#ab85546, Abcam plc.), diluted at 1:100. Subsequently, the secondary antibody (#sc-2023; Santa Cruz Biotechnology, Inc.), diluted at 1:200 in 1.5% donkey serum, was applied for 30 min at room temperature.
Sc 2023
Manufactured by Santa Cruz Biotechnology
Sourced in United Kingdom
Sc-2023 is a laboratory equipment product from Santa Cruz Biotechnology. It is designed for general laboratory use. The core function of Sc-2023 is to provide a reliable and consistent platform for various scientific applications. Further details on the specific intended use of this product are not available.
Automatically generated - may contain errors
Lab products found in correlation
Axiovert 100 microscope, by Leica (1 mentions)
Ab97051, by Abcam (1 mentions)
Ab181547, by Abcam (1 mentions)
Ab150061, by Abcam (1 mentions)
Ab175472, by Abcam (1 mentions)
Ab172730, by Abcam (1 mentions)
P36931, by Thermo Fisher Scientific (1 mentions)
Sc 1190, by Santa Cruz Biotechnology (1 mentions)
4 protocols using sc 2023
1
XBP1 Immunofluorescent Staining
2
Histological Analysis of Pancreatic Islets
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a) H&E Staining: pictures of whole tissue slides and of every islet of Langerhans were taken using the Zeiss Axiovert 100 microscope (25x/200x) and the Leica EC3 digital camera. The islets were counted, and the islet area was measured using ImageJ software to calculate the islet density and the mean islet area per slide.
b) Pancreas immunohistology: The beta cell content of islets of Langerhans was measured using immunohistological staining of insulin. We used an antibody against insulin (1:200, ab181547, abcam, Cambridge, United Kingdom) and a secondary antibody (1:500, ab97051, abcam, Cambridge, United Kingdom) diluted in antibody diluent (Dako, Glostrup, DK), and for visualization the ABC staining system (sc 2023, Santa Cruz Biotechnology, Santa Cruz, CA) following the instructions provided by the manufacturer. All islets per slide were photographed using an Olympus BH-2 microscope (200x) and a CFW-1310C digital camera. Thirty images for each sample were taken. The islet area and beta cell content were measured using ImageJ. The average islet size was obtained by total islet area/islet amount in each sample. The beta cell content was determined by the insulin positive staining area.
3
Immunofluorescence analysis of sperm proteins
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Dry spermatozoa smeared on poly-L-lysine-coated slides were fixed in acetone:methanol (1:1) solution at 4 °C for 10 min. Slides then were blocked in 5% donkey serum in 0.1% Triton X-100 PBS for 1 h and incubated with GSK3α/β, ATP5A, and cyclophilin D antibodies mixed in 1.5% donkey serum in a humidified chamber overnight at 4˚C. Rabbit IgG (ab172730; Abcam) and mouse IgG (sc-2023; Santa Cruz) were used as negative controls. After washing three times in PBS, slides were incubated with Alexa Fluor 488-conjugated anti-rabbit IgG (ab150061; Abcam) and Alexa Fluor 568-conjugated anti-mouse IgG (ab175472; Abcam) mixed in 1.5% donkey serum PBS for 1 h at room temperature. After washing three times in PBS, slides were mounted with DAPI (P36931; Invitrogen). Fluorescence images were captured by a microscope system with a cooled CCD (DP71; Olympus, Tokyo, Japan).
4
Immunohistochemical Evaluation of Cancer Cell Markers
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Thin sections of 10 % formalin-fixed, paraffin-embedded tissue specimens were treated with goat anti-human RPN2 antibody (sc-12165, Santa Cruz), rabbit anti-human HMGB1 antibody (#6893S, Cell Signaling), or rabbit anti-human NFKB1 antibody (sc-1190, Santa Cruz), followed by a peroxidase-conjugated goat anti-rabbit (sc-2018, Santa Cruz) or rabbit anti-goat (sc-2023, Santa Cruz) secondary antibody. Color was developed using Avidin and Biotin-conjugated horseradish peroxidase (ABC reagents) and according to standard protocols. The percentage of positively stained cancer cells was determined under the microscope from more than four visual fields (at 400× magnification). Specimens were evaluated by two independent pathologists and classified into 2 groups: negative staining (no cells were intensely stained), and positive staining (at least 10 % cells were intensely stained) [30 (link)].
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