Pancreas was fixed in 4% paraformaldehyde, embedded in paraffin, and sectioned by Histology Core Facility at Cornell and University of Michigan. For Hematoxylin/eosin staining, ten-micrometer-thick paraffin sections were stained and imaged using Aperio Scanscope. For immunostaining, paraffin-embedded sections were rehydrated with sequential wash in xylene, 100%, 95%, 75% ethanol and water and boiled in 1 mM EDTA for antigen retrieval. Subsequently, sections were blocked using 5% donkey serum and incubated at 4 °C overnight with primary antibodies: Anti-insulin (Abcam ab7842 or Linco 4011, 1:200), glucagon (Sigma K79bB10, 1:1000), Ki-67 (Abcam ab15580, 1:50), Pcna (Santa Cruz sc-56, 1:100), CD31 (Santa Cruz sc-1506, 1:100), Pdx1 (Cell Signaling D59H3, 1:100), Cdk4 (Santa Cruz sc-260, 1:100) and Ccnd2 (Santa Cruz sc-593, 1:100). Following day, slides were washed and incubated with conjugated secondary antibodies and mounted on slides with prolong gold antifade/ DAPI for nuclear staining. Fluorescence Images were captured under a Nikon A1 confocal microscope at Brehm Diabetes Research Center Imaging Facility at University of Michigan. For horseradish peroxidase enzyme (HRP) staining, slides were stained with Histostain kit and DAB substrate from Invitrogen.
Sc 593
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Sc-593 is a lab equipment product manufactured by Santa Cruz Biotechnology. It is designed for use in various scientific research applications. The core function of Sc-593 is to facilitate the specific task it is intended for, but a detailed description cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Dab substrate, by Thermo Fisher Scientific (2 mentions)
Histostain kit, by Thermo Fisher Scientific (2 mentions)
Ab7842, by Abcam (2 mentions)
Ab15580, by Santa Cruz Biotechnology (2 mentions)
A1 confocal microscope, by Nikon (2 mentions)
Linco 4011, by Merck Group (2 mentions)
K79bb10, by Abcam (2 mentions)
Sc 56, by Santa Cruz Biotechnology (2 mentions)
Sc 1506, by Cell Signaling Technology (2 mentions)
Sc 260, by Santa Cruz Biotechnology (2 mentions)
α gata4, by Santa Cruz Biotechnology (1 mentions)
α foxl2, by Novus Biologicals (1 mentions)
α smad3, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 555, by Thermo Fisher Scientific (1 mentions)
Prolong gold antifade reagent with dapi, by Thermo Fisher Scientific (1 mentions)
Sc 528, by Santa Cruz Biotechnology (1 mentions)
Sp5 laser scanning confocal microscope, by Leica (1 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (1 mentions)
Ab13840, by Abcam (1 mentions)
Sc 1641, by Santa Cruz Biotechnology (1 mentions)
4 protocols using sc 593
1
Histological Analysis of Pancreatic Tissue
2
Histological Analysis of Pancreatic Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Pancreas was fixed in 4% paraformaldehyde, embedded in paraffin, and sectioned by Histology Core Facility at Cornell and University of Michigan. For Hematoxylin/eosin staining, ten-micrometer-thick paraffin sections were stained and imaged using Aperio Scanscope. For immunostaining, paraffin-embedded sections were rehydrated with sequential wash in xylene, 100%, 95%, 75% ethanol and water and boiled in 1 mM EDTA for antigen retrieval. Subsequently, sections were blocked using 5% donkey serum and incubated at 4 °C overnight with primary antibodies: Anti-insulin (Abcam ab7842 or Linco 4011, 1:200), glucagon (Sigma K79bB10, 1:1000), Ki-67 (Abcam ab15580, 1:50), Pcna (Santa Cruz sc-56, 1:100), CD31 (Santa Cruz sc-1506, 1:100), Pdx1 (Cell Signaling D59H3, 1:100), Cdk4 (Santa Cruz sc-260, 1:100) and Ccnd2 (Santa Cruz sc-593, 1:100). Following day, slides were washed and incubated with conjugated secondary antibodies and mounted on slides with prolong gold antifade/ DAPI for nuclear staining. Fluorescence Images were captured under a Nikon A1 confocal microscope at Brehm Diabetes Research Center Imaging Facility at University of Michigan. For horseradish peroxidase enzyme (HRP) staining, slides were stained with Histostain kit and DAB substrate from Invitrogen.
3
Immunohistochemical Analysis of GCT Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
GCT tissue microarray and normal ovarian samples were stained as previously described and presented for GATA4 and CCND2 [25] (link), [26] (link), with following primary antibodies: α-GATA4 1∶400 (sc-1237, Santa Cruz Biotechnology, Santa Cruz, CA, USA), α-FOXL2 1∶400 (IMG-3228, Imgenex, San Diego, CA, USA), α-SMAD3 1∶400 (#51–1500, Invitrogen Corporation, Carlsbad, CA, USA), and α-CCND2 1∶1000 (sc-593; Santa Cruz Biotechnology). The GCT stainings were classified into three groups based on the intensity of staining: high representing intermediary or strong intensity in 70% of the cells, intermediate representing moderate or strong intensity in <70% of the cells, and low representing negative or low intensity even if detected in 100% of the cells; scoring of GATA4 staining as described (34), i.e. high for 80–100% positive nuclei, intermediate for 20–80% positive nuclei, and low for 0–20% positive nuclei. Two researchers (M.A. and N.A. or A.F) independently performed the evaluation and disagreements were resolved by a joint review.
4
Immunofluorescent Localization of SMAD Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Sections were dewaxed before microwaving in 0.1 M citrate buffer (pH6.0). Sections were blocked with CAS universal blocking reagent (ThermoFisher) for 20 minutes at room temperature to reduce non-specific binding. Specific antibodies against SMAD2 (0.59 µg/ml, #5339, Cell Signaling Technology, MA, USA), SMAD3 (0.75 µg/ml, #9523, Cell Signaling), SMAD2/3 (0.25 µg/ml, 133098, Santa Cruz Biotechnology, TX, USA), CCND2 (0.5 µg/ml, sc-593, Santa Cruz Biotechnology, TX, USA), DDX4 (2.2 µg/ml, ab13840, Abcam, Cambridge, UK) and P27 (1 µg/ml, sc-528, Santa Cruz or 0.5 µg/ml, sc-1641 for co-localisation) were diluted in blocking solution and incubated overnight at 4 °C. A species-specific isotype control IgG antibody at the same concentration (normal rabbit IgG I-1000 and normal mouse IgG I-200, Vector Laboratories) were included in each experiment. Sections were washed in PBS and incubated with secondary antibodies (either donkey anti-rabbit AlexaFluor® 488 or AlexaFluor® 555; donkey anti-mouse AlexaFluor® 488 and AlexaFluor® 555, each 1:400; Invitrogen). Sections were mounted with a drop of Prolong® Gold anti-fade reagent with DAPI (ThermoFisher) and were imaged using an inverted Leica SP5 confocal laser-scanning microscope (Leica Microsystems, Wetzlar, Germany).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!