Immunofluorescent staining was performed as previously published [22 (link)] with modifications to account for the use of paraffin embedded tissue. Briefly, tissue sections were deparaffinized through graded ethanols (100%, 95%, 85%, 75%, 50%; Fisher Scientific) and then blocked using normal donkey serum (Sigma). Primary antibodies used included guinea pig anti-insulin (DAKO; 1:500), mouse anti-glucagon (Abcam; 1:500), rat anti-somatostatin (abcam; 1:200), goat anti-pancreatic polypeptide (abcam; 1:200), goat anti-ghrelin (Santa Cruz; 1:500), mouse anti-Pax5 (DAKO; 1:200), mouse anti-CD8 (Thermo Fisher; 1:500), mouse anti-CD4 (Leica; 1:500), rabbit anti-eIF5AHyp ([13 (link),18 (link)]; 1:1000). Secondary antibodies including Alexa-488, Cy3, or Alexa-647 (Jackson Immunoresearch) were used to visualize primary antibodies. DAPI (Sigma; 1:1000) was used to visualize nuclei. Images were acquired with a Zeiss 710 confocal microscope.
Goat anti ghrelin
Manufactured by Santa Cruz Biotechnology
Goat anti-ghrelin is an antibody product designed for research applications. It is produced in goats and is reactive against the ghrelin protein. The core function of this product is to specifically bind and detect the ghrelin protein in various experimental procedures.
Automatically generated - may contain errors
Lab products found in correlation
710 confocal microscope, by Zeiss (3 mentions)
Normal donkey serum (nds), by Merck Group (1 mentions)
Alexa 647, by Jackson ImmunoResearch (1 mentions)
Goat anti pancreatic polypeptide, by Abcam (1 mentions)
Mouse anti glucagon, by Abcam (1 mentions)
Mouse anti cd4, by Leica camera (1 mentions)
Guinea pig anti insulin, by Agilent Technologies (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Cyanine3 (cy3), by Jackson ImmunoResearch (1 mentions)
Alexa 488, by Jackson ImmunoResearch (1 mentions)
Cas block, by Thermo Fisher Scientific (1 mentions)
Tcs sp8 confocal microscope, by Nikon (1 mentions)
Eclipse 80i widefield microscope, by Nikon (1 mentions)
Anti guinea pig cy5, by Jackson ImmunoResearch (1 mentions)
Anti goat cy3, by Jackson ImmunoResearch (1 mentions)
2 protocols using goat anti ghrelin
1
Immunofluorescent Staining of Paraffin Embedded Tissue
2
Islet Cell Immunofluorescence Staining
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Whole islets or trypsin-dispersed islet cells were fixed in 4% paraformaldehyde, then washed with phosphate-buffered saline (PBS). Dispersed islet cells were attached to positively-charged slides using a CytoSpin. Islets were incubated in blocking solution (PBS with 0.3% Triton and 10% fetal bovine serum [FBS]), and slides in CAS-Block (ThermoFisher), prior to and concurrent with antibodies. Primary antibodies (rabbit anti-GC [Abcam ab65636], rabbit anti-CHODL [Abcam ab134924], mouse anti-glucagon [Abcam ab82270], guinea pig anti-insulin [Invitrogen 180067], rabbit anti-somatostatin [Santa Cruz sc-13099], goat anti-pancreatic polypeptide [PP; Abcam ab77192], and goat anti-ghrelin [Santa Cruz sc-10368]; all 1:100) were incubated overnight at 4 °C, and secondary antibodies (anti-rabbit Cy2 or Cy3, anti-mouse Cy3 or Cy5, anti-goat Cy3, and anti-guinea pig Cy5 [Jackson ImmunoResearch]; all 1:200) were incubated at room temperature for 3 h. DAPI (4′,6-diamidino-2-phenylindole) was added with the mounting media to counterstain DNA. Whole islets were imaged on a Leica TCS SP8 confocal microscope, and dispersed cells were imaged on a Nikon Eclipse 80i widefield microscope.
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