Immunostaining Protocol for Gland Samples

Salivary glands or larval samples were made exactly as mentioned in our previous work (Ghosh et al, 2019 (link); Mathre et al, 2019 (link)). Dilutions of antibodies used: 1:4,000 for anti-tubulin (E7-c), (mouse) from DSHB, 1:1,000 for anti-actin antibody (Cat# A5060) (Rabbit) from Sigma-Aldrich, 1:2,000 for anti-GFP antibody (Cat# sc-9996) (Rabbit) from Santa Cruz, 1:1,000 for anti-mCherry antibody (Cat# PA5-34974) (Rabbit) from Thermo Fisher Scientific, 1:1,000 for anti-HA antibody (Cat# 2367S) (Mouse) from CST and Normal Rabbit IgG (sc-2027) from Santa Cruz, 1:1,000 for anti-dPIP4K (generated for the laboratory by NeoBiolab).

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Ghosh A., Venugopal A., Shinde D., Sharma S., Krishnan M., Mathre S., Krishnan H., Saha S, & Raghu P. (2023). PI3P-dependent regulation of cell size and autophagy by phosphatidylinositol 5-phosphate 4-kinase. Life Science Alliance, 6(9), e202301920.

Publication 2023

anti-actin antibody anti-GFP antibody anti-mCherry antibody Normal Rabbit IgG

Actin Anti antibody Antibodies Dilutions Larval Mouse Rabbit Salivary glands Tubulin

Corresponding Organization : National Centre for Biological Sciences

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Variable analysis

independent variables

Dilutions of antibodies

dependent variables

Salivary glands or larval samples

control variables

Salivary glands or larval samples were made exactly as mentioned in our previous work (Ghosh et al, 2019; Mathre et al, 2019)

controls

Positive control: None specified
Negative control: Normal Rabbit IgG (sc-2027) from Santa Cruz

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