Dissection, staining, and mounting of wandering third instar larvae was performed as previously described (Sulkowski et al., 2011 (link)). For immunohistochemistry (IHC), larvae were dissected in PBS, pinned on sylgard plates and fixed in 4% paraformaldehyde for 20 minutes. Next, larvae were washed five times in 1X PBT (PBS + 0.3% Triton X-100). Cuticle filets were blocked in 5% normal donkey serum (Jackson Laboratories, West Grove, PA) for at least 30 minutes at room temperature followed by incubation with respective primary antibodies. Antibody dilutions used were as follows: rabbit anti-Bdwf (1:200), mouse anti-CD8 (1:100) (Invitrogen), rabbit RpS6-(1:100) (Cell Signaling Technology), and mouse anti-Cut (2B10; 1:50) (Developmental Studies Hybridoma Bank). Donkey anti-rabbit and donkey anti-mouse secondary antibodies were used at 1:200 (Jackson Immunoresearch). Filets were incubated in glycerol for 5 minutes, followed by being directly mounted onto coverslips with either a drop of glycerol, or aqueous fluoromount. The slides were then imaged on a Nikon C1 Plus confocal microscope or Zeiss LSM 780.
Rabbit rps6
Manufactured by Cell Signaling Technology
Sourced in United States
Rabbit RpS6 is a primary antibody that recognizes the ribosomal protein S6 from rabbit species. Ribosomal protein S6 is a component of the 40S subunit of the eukaryotic ribosome and plays a role in the regulation of cell growth and proliferation.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 780, by Zeiss (4 mentions)
Normal donkey serum (nds), by Jackson ImmunoResearch (2 mentions)
Rabbit anti bdwf, by Thermo Fisher Scientific (2 mentions)
Anti mouse cd8, by Thermo Fisher Scientific (2 mentions)
C1 plus confocal microscope, by Zeiss (2 mentions)
Mouse anti cut 2b10, by Developmental Studies Hybridoma Bank (2 mentions)
Donkey anti mouse secondary antibody, by Jackson ImmunoResearch (1 mentions)
2 protocols using rabbit rps6
1
Larval Dissection and Immunohistochemistry
2
Larval Dissection and Immunostaining
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Dissection, staining, and mounting of wandering third-instar larvae were performed as previously described [19 (link)]. For immunohistochemistry (IHC), larvae were dissected in PBS, pinned on Sylgard plates, and fixed in 4% paraformaldehyde for 20 minutes. Next, larvae were washed five times in 1× PBT (PBS + 0.3% Triton X-100). Cuticle filets were blocked for at least 30 minutes at room temperature in 5% normal donkey serum (Jackson Laboratories, West Grove, PA, USA), followed by incubation with respective primary antibodies. Antibody dilutions used were as follows: rabbit anti-Bdwf (1:200), mouse anti-CD8 (1:100) (Invitrogen), rabbit RpS6-(1:100) (Cell Signaling Technology, Danvers, MA, USA), and mouse anti-Cut (2B10; 1:50) (Developmental Studies Hybridoma Bank, Houston, TX, USA). Donkey anti-rabbit and donkey anti-mouse secondary antibodies were used at 1:200 (Jackson Immunoresearch, West Grove, PA, USA). Filets were incubated in glycerol for 5 minutes, followed by being directly mounted onto coverslips with either a drop of glycerol or an aqueous fluoromount. The slides were then imaged on a Nikon C1 Plus confocal microscope or a Zeiss LSM 780.
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