Immunostaining Techniques for Drosophila Proteins

For immunostaining, the following primary antibodies were used: rabbit anti-CENP-A (CID) antibody (active motif 39719; 1:500), sheep anti-CENP-C (Dattoli et al., 2020 (link)); 1:2000), rabbit anti-VASA (Santa Cruz sc-30210; 1:500), rat anti-VASA (Developmental Studies Hybridoma Bank [DSHB]; 1:500), sheep anti-CAL1 (this study; 1:1000), rabbit anti-SMAD3/5 (pMad; Abcam ab52903; 1:500), DAPI (1:1000), mouse anti-Armadillo (DSHB N2 7A1; 1:700), mouse anti-1B1 (Hts; DSHB; 1:100), mouse anti-BAM (DSHB; 1:50), mouse anti-FIBRILLARIN (Abcam; ab5821; 1:500), mouse anti-HA (ThermoFisher; 26183; 1:500), mouse anti-gamma-tubulin (Abcam; ab44928; 1:500), mouse anti-FASCILLIN III (DSHB 7G10; 1:200), rat anti-mCherry (Chromotek 5F8; 1:200). Sheep anti-CAL1 antibody was raised against CAL1 amino acids 748–979 with a C-terminal His tag produced according to Bade et al. (2014 (link)). Species-specific Alexa Fluor 405, 488, 546, and 647 secondary antibodies were used at a 1:500 dilution.

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Kochendoerfer A.M., Keegan R.S, & Dunleavy E.M. (2023). Centromere proteins are asymmetrically distributed between newly divided germline stem and daughter cells and maintain a balanced niche in Drosophila males. Molecular Biology of the Cell, 34(5), ar42.

Publication 2023

sc-30210 rat anti-VASA ab52903

Amino acids Anti antibody Antibodies Antibody Armadillo Bade Cenp c Dapi Dilution Fibrillarin Gamma tubulin Hybridoma Mouse Rabbit Sheep Smad3

Corresponding Organization :

Other organizations : Ollscoil na Gaillimhe – University of Galway

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Variable analysis

independent variables

Rabbit anti-CENP-A (CID) antibody (active motif 39719; 1:500)
Sheep anti-CENP-C (Dattoli et al., 2020 (link); 1:2000)
Rabbit anti-VASA (Santa Cruz sc-30210; 1:500)
Rat anti-VASA (Developmental Studies Hybridoma Bank [DSHB]; 1:500)
Sheep anti-CAL1 (this study; 1:1000)
Rabbit anti-SMAD3/5 (pMad; Abcam ab52903; 1:500)
Mouse anti-Armadillo (DSHB N2 7A1; 1:700)
Mouse anti-1B1 (Hts; DSHB; 1:100)
Mouse anti-BAM (DSHB; 1:50)
Mouse anti-FIBRILLARIN (Abcam; ab5821; 1:500)
Mouse anti-HA (ThermoFisher; 26183; 1:500)
Mouse anti-gamma-tubulin (Abcam; ab44928; 1:500)
Mouse anti-FASCILLIN III (DSHB 7G10; 1:200)
Rat anti-mCherry (Chromotek 5F8; 1:200)

dependent variables

Not explicitly mentioned

control variables

DAPI (1:1000)

controls

Positive controls: Not specified
Negative controls: Not specified

Annotations

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