Discs were dissected from wandering third instar larvae and immunofluorescence staining was carried out as previously described [33 (link)]. Briefly, larvae are dissected in cold PBS (pH7.4) and fixed in 4% formaldehyde in PBS for 15 minutes. Samples were then permeabilized in PBS + 0.1% Triton X-100 and blocked in 5% normal horse serum. Primary antibodies in 5% normal horse serum were incubated overnight at 4°C or 2 hours at RT. Primary antibodies used for the immunostainings were mouse or rabbit α-V5 (Life Technology), rabbit anti-pMad (1:200) (Cell Signaling), rabbit α-Spalt major (1:100), anti-hedgehog (1:40), and chicken α-lacZ (1:100). Mouse anti-patched (DSHB), mouse anti-engrailed (DSHB), rat anti-Ci 2A1 (DSHB) were obtained from Developmental Studies Hybridoma Bank (DSHB), University of Iowa, Department of Biological Sciences. Fluorophore conjugated secondary antibodies were from Jackson Immunoresearch. Confocal images were collected as single frames on a Zeiss LSM 780 confocal microscope. MG132 (100 μM; Calbiochem) in M3 medium (Sigma) was used to treat wing discs for up to 6 hours before immunostaining.
Mouse anti patched
Manufactured by Developmental Studies Hybridoma Bank
Sourced in Israel
Mouse anti-patched is a laboratory reagent produced by the Developmental Studies Hybridoma Bank. It is an antibody that specifically binds to the patched protein in mice. The patched protein is a key component of the hedgehog signaling pathway, which is involved in embryonic development and tissue homeostasis.
Automatically generated - may contain errors
Lab products found in correlation
Fluorophore conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions)
Lsm 780 confocal microscope, by Zeiss (1 mentions)
M3 medium, by Merck Group (1 mentions)
Rabbit anti pmad, by Cell Signaling Technology (1 mentions)
Rat anti ci 2a1, by Developmental Studies Hybridoma Bank (1 mentions)
Mouse anti engrailed, by Developmental Studies Hybridoma Bank (1 mentions)
Rabbit anti phospho histone h3, by Merck Group (1 mentions)
Tcs sp8, by Leica (1 mentions)
Rabbit anti gfp, by Abcam (1 mentions)
Rat anti ci, by Developmental Studies Hybridoma Bank (1 mentions)
Mouse anti egfr, by Abcam (1 mentions)
2 protocols using mouse anti patched
1
Immunofluorescence Staining of Drosophila Wing Discs
2
Dissection and Imaging of Eye-Antenna Discs
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Eye-antenna discs were dissected, fixed and stained as described previously25 (link)26 (link). Tumour and adult eyes sizes analyses were carried out on a Leica MZ FLIII fluorescence stereomicroscope equipped with a camera. Samples were examined by confocal microscopy with a Zeiss LSM510 Meta system and a Leica TCS SP8 confocal microscope. Images were analysed and processed with IMARIS (Bitplane, Switzerland) and Illustrator (Adobe) software, respectively. The following primary antibodies were used: guinea pig anti-Hrs (1:2,000, H. Bellen); Mouse anti-Spi (1:500; A.D. Vrailas-Mortimer); Rabbit anti-phosphohistone-H3 (1:1,000, Sigma); Rat anti-DER (1:1,000, B. Shilo, Weizmann Institute of Science/Israel); Mouse anti-EGFR (1:500, abcam); Rabbit anti-GFP (1:1,000, Abcam); Rabbit anti-Hh (1:500 Pre-absorbed, T. Kornberg, UCSF); Rat anti-Ci (detects the full length/active version of Ci, 1:200, Developmental Studies Hybridoma Bank, 2A1); Mouse anti-Patched (1:200, Developmental Studies Hybridoma Bank, Apa1). Secondary antibodies were from Invitrogen. TUNEL staining was performed using the Apoptag Red kit from Chemicon.
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