Immunofluorescence of Pancreatic Tissue

Immunofluorescence was performed as previously described [20 (link)]. Pancreatic tissue was fixed in 10% neutral buffered formalin and antigen retrieval was completed with Antigen Unmasking Solution (Vector Laboratories). Primary antibodies included ARC (Cayman), Ki67 (Abcam), and insulin (Abcam), and secondary antibodies were Alexa Fluor 488 or 568 (Invitrogen). Samples were counterstained and coverslipped with VECTASHIELD HardSet Mounting Medium with DAPI (Vector Laboratories).

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McKimpson W.M., Yuan Z., Zheng M., Crabtree J.S., Libutti S.K, & Kitsis R.N. (2015). The Cell Death Inhibitor ARC Is Induced in a Tissue-Specific Manner by Deletion of the Tumor Suppressor Gene Men1, but Not Required for Tumor Development and Growth. PLoS ONE, 10(12), e0145792.

Publication 2015

Antigen Unmasking Solution insulin Alexa Fluor 488 VECTASHIELD HardSet Mounting Medium with DAPI

Alexa fluor 488 Antibodies Antigen Cayman Dapi Formalin Immunofluorescence Insulin Pancreatic Tissue Vector

Corresponding Organization : Louisiana State University Health Sciences Center New Orleans

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Variable analysis

independent variables

Antigen retrieval method (Antigen Unmasking Solution)

dependent variables

Immunofluorescence signal intensity
Presence and localization of ARC, Ki67, and insulin proteins

control variables

Tissue type (pancreatic tissue)
Fixation method (10% neutral buffered formalin)
Primary antibodies (ARC, Ki67, insulin)
Secondary antibodies (Alexa Fluor 488 or 568)
Counterstaining and mounting medium (VECTASHIELD HardSet Mounting Medium with DAPI)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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