Human pancreatic paraffin sections were used for IF co-localization analyses. Sections were sequentially stained with rabbit anti-IL18 (1:25, ab207324, Abcam, Cambridge, MA) and mouse anti-glucagon (1:200, 14-9743-82, eBioscience, San Diego, CA) antibodies; Rabbit anti-IL18r (1:50, HPA007615, Sigma-Aldrich) and mouse anti-amylase 1A (1:100, SAB4200673, Sigma-Aldrich) antibodies; rabbit anti-NCC (1:50, AB3553, Millipore, Burlington, MA) and rat anti-insulin (1:500, MAB 1417, R&D Systems); rabbit anti-NCC (1:50, AB3553, Millipore) and mouse anti-glucagon (1:200, 14-9743-82, eBioscience); rabbit anti-NCC (1:50, AB3553, Millipore) and rat anti-Mac-2 (1:200, CL8942LE, Cedarlane Laboratories, Burlington, Canada) antibodies, followed by Alex Fluor 555, 488-labeled secondary antibody detections (Thermo Fisher Scientific, Waltham, MA). Slides were counterstained with DAPI (R37606, Invitrogen, San Diego), and images captured with a confocal microscopy (Olymbus Fluoview FV1000; Olymbus). Five random fields from each section were examined, and the data were presented as positive area percentage by detecting the staining intensity with the Image-Pro Plus software.
Mab1417
Manufactured by R&D Systems
Sourced in United States
MAB1417 is a monoclonal antibody produced by R&D Systems. It is intended for use in immunoassays to detect and quantify human Interleukin-23 (IL-23).
Automatically generated - may contain errors
Lab products found in correlation
Ab92517, by Abcam (2 mentions)
Ab207324, by Abcam (1 mentions)
R37606, by Thermo Fisher Scientific (1 mentions)
Cl8942le, by Cedarlane (1 mentions)
Hpa007615, by Merck Group (1 mentions)
Ab3553, by Merck Group (1 mentions)
Ab47383, by Abcam (1 mentions)
Ab9361, by Abcam (1 mentions)
Ab57222, by Abcam (1 mentions)
Hpa005653, by Merck Group (1 mentions)
H 031 31, by Phoenix Pharmaceuticals (1 mentions)
A0566, by Agilent Technologies (1 mentions)
Ultraab diluent, by Thermo Fisher Scientific (1 mentions)
Alexa 488, by Thermo Fisher Scientific (1 mentions)
Alexa 568, by Thermo Fisher Scientific (1 mentions)
G2654, by Merck Group (1 mentions)
Lsm 510 confocal microscopy, by Zeiss (1 mentions)
Fluoroshield with dapi, by Merck Group (1 mentions)
A21206, by Thermo Fisher Scientific (1 mentions)
A11007, by Thermo Fisher Scientific (1 mentions)
5 protocols using mab1417
1
Immunofluorescent Localization of Pancreatic Proteins
2
Comprehensive Immunofluorescence Staining Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunofluorescence was performed as described previously (26) . Rabbit anti-insulin (20056,1:1,000; Immuno-Star), guinea pig anti-insulin (4011-01F,1:100; LINCO), rat anti-insulin (MAB1417, 1:500; R&D Systems), rabbit antiglucagon (20076,1:2,000; ImmunoStar), guinea pig antiglucagon (1:500; LINCO), rabbit anti-somatostatin (A0566 1:500, DakoCytomation; and HPA019472, 1:2,500, Sigma-Aldrich), rabbit anti-PP (AB939,1:500; Chemicon International), chicken anti-b-galactosidase (b-gal) (ab9361,1:1,000; Abcam), mouse and rabbit anti-ghrelin (ab57222, 1:100, Abcam; and H-031-31, 1:1,000, Phoenix Pharmaceuticals), rabbit anti-MafA (34) , rabbit anti-MafB (HPA005653,1:5,000; Sigma-Aldrich), goat anti-Pdx1 (ab47383,1:5,000; Abcam), mouse anti-Isl1/2 (39.4D5, 1:50; DSHB), mouse anti-Ngn3 (F25A1B3, 1:50; DSHB), mouse anti-Pax6 (PAX6-S, 1:20; DSHB), mouse anti-Nkx6.1 (F55A10-C, 1:200; DSHB), and rat anti-Ki67 (14-5698, 1:50; Invitrogen) were used as primary antibodies. Secondary antibodies coupled to Cy3, Cy2, or Cy5 against rabbit, rat, mouse, chicken, or guinea pig (Jackson ImmunoResearch Laboratories) were used. Fluorescence imaging was performed using a Leica confocal microscope, and image processing was performed using Adobe Photoshop software.
3
Immunofluorescent Staining of Human Islets
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Briefly, human islets were stained with a polyclonal rabbit antibody against GPR44 (HPA014259, dilution 1:10, Atlas Antibodies, Stockholm, Sweden), as well as with rat anti-human insulin (MAB1417, dilution 1:100, R&D Systems, Minneapolis, MN, USA) and Alexa-647-conjugated mouse anti-glucagon (G2654, dilution 1:500, Sigma-Aldrich). Primary antibodies were diluted in UltraAb Diluent (Thermo Fisher Scientific, Fermont, CA, USA) and incubated with the islets for 24 h at 4 °C. The GPR44 antibody was visualized by a secondary antibody conjugated with Alexa-568 and the insulin antibody by a secondary antibody conjugated with Alexa-488 (Dilution 1:500, Invitrogen, Carlsbad, CA, USA). Secondary conjugated antibodies were incubated with the islets for 24 h at 4 °C. The islets were then mounted with Fluoroshield with DAPI (Sigma-Aldrich) and analyzed with Zeiss LSM 510 confocal microscopy (Carl Zeiss, Jena, Germany). A detail of the procedures was described previously [2 (link)].
4
Morphological Analysis of KKAy Mouse Islets
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The fixed pancreases of KKAy mice were embedded in paraffin. Tissues were sliced to 5 μm for morphology analyses. Some sections were stained with hematoxylin-eosin to observe the morphology of islets under a microscope. Others were immunostained with rabbit anti-glucagon antibody (1:200, ab92517, Abcam, USA), rabbit anti-ALDH1A3 antibody (1:100, NBP2-15339, NOVUS, USA), and rat anti-insulin antibody (1:200, MAB1417, R&D Systems, USA). The secondary antibodies were Alexa Fluor 488 donkey anti-rabbit IgG (1:200, A-21206, Invitrogen, USA) and Alexa Fluor 594 donkey anti-rat IgG (1:200, A-11007, Invitrogen, USA). Then, the tissues were mounted utilizing mounting medium with DAPI (ZLI-9557, ZSGB-BIO, China). Finally, the tissues were visualized by confocal laser scanning microscopy (CLSM) (Leica Microsystems, Germany). ImageJ software was used to analyze the picture. The area percentage of β-cell/islet and α-cell/islet were calculated via the insulin-positive or glucagon-positive area in one islet divided by the islet area. The ratio of ALDH1A3-positive β cells/β cells were calculated via the both ALDH1A3 and insulin-positive area divided by insulin-positive area in the islet.
5
Quantitative Analysis of Pancreatic Islets
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All mice were sacrificed through cervical dislocation and the pancreas were fixed in 10% formalin, embedded in paraffin, and dissected to prepare 5-μm-thick slides, then stained with antibodies against insulin (MAB1417; R&D Systems, USA) and glucagon (ab92517; Abcam, USA) as previously described (Li et al., 2017 (link)). The mean fluorescence intensities of insulin and glucagon, which account for islets areas were quantified using Image J software.
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!