The following primary antibodies were used: guinea-pig anti-Tj (G5 or GP6, 1:5000) [57 (link)], rat anti-Bab2 (R10, 1:3000; or R7, 1:2000) [20 (link)], rabbit anti-Vasa (1:2000) [80 (link)], rabbit anti-Vasa (d-260, 1:500; Santa Cruz Biotechnology), chicken anti-Vasa (1:5000; gift from K. Howard and M. Van Doren), rabbit anti-α-spectrin (#254, 1:1000; gift from D. Branton), mouse anti-LamC (LC28.26, 1:50), mouse anti-Hts (1B1, 1:5), mouse anti-N (C17.9C6, 1:5; C458.2H, 1:5), mouse anti-Dl (C594.9B, 1:5), mouse anti-Engrailed (4D9, 1:5), and mouse anti-ßPS integrin (CF.6G11, 1:10) (Developmental Studies Hybridoma Bank), rabbit anti-pMad (PS1, 1:250; gift from T. Tabata) [81 (link)], rabbit anti-pMad (pSmad1/5, 41D10, 1:100; Cell Signalling), rabbit anti-ß-galactosidase (1:1500; MP Biomedicals), and rabbit anti-GFP (1:100; BD Biosciences). Secondary antibodies (1:400) were conjugated either to Cy3, Cy5 (Jackson Immuno Research Laboratories), Alexa-405, Alexa-555, Alexa-488, or Alexa-647 (Molecular Probes, Life Technologies). Ovaries were mounted in Vectashield (Vector Laboratories).
Mouse anti dl
Manufactured by Developmental Studies Hybridoma Bank
Sourced in United States
Mouse anti-Dl is a monoclonal antibody that specifically binds to the Delta protein. The Delta protein is a transmembrane ligand that plays a key role in the Notch signaling pathway, which is important for cell-cell communication and developmental processes. This antibody can be used to detect and study the Delta protein in various experimental systems.
Automatically generated - may contain errors
Lab products found in correlation
Alexa 488, by Thermo Fisher Scientific (1 mentions)
Alexa 647, by Thermo Fisher Scientific (1 mentions)
Alexa 405, by Thermo Fisher Scientific (1 mentions)
Alexa 555, by Thermo Fisher Scientific (1 mentions)
Rabbit anti vasa d 260, by Santa Cruz Biotechnology (1 mentions)
Vectashield, by Vector Laboratories (1 mentions)
Mouse anti engrailed, by Developmental Studies Hybridoma Bank (1 mentions)
Rabbit anti gfp, by BD (1 mentions)
Cyanine3 (cy3), by Jackson ImmunoResearch (1 mentions)
Alexa fluor dye, by Thermo Fisher Scientific (1 mentions)
Phalloidin tritc, by Merck Group (1 mentions)
F actin, by Merck Group (1 mentions)
Phalloidin atto 647n, by Merck Group (1 mentions)
Anti dlg, by Developmental Studies Hybridoma Bank (1 mentions)
Dapi staining, by Thermo Fisher Scientific (1 mentions)
Lsm 700, by Zeiss (1 mentions)
Ab13970, by Abcam (1 mentions)
Dapi dye, by Merck Group (1 mentions)
A11034, by Thermo Fisher Scientific (1 mentions)
Mouse anti β gal, by Developmental Studies Hybridoma Bank (1 mentions)
6 protocols using mouse anti dl
1
Drosophila Ovary Immunohistochemistry
2
Immunolabeling and Proliferation Assay of Drosophila Midgut
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Adult midguts were dissected in PBS and fixed for 30 min at room temperature in 4% (v/v) paraformaldehyde in PEM (0.1 M PIPES, 2 mM EGTA, 1 mM MgSO4). Triton-X and sodium deoxycholate were added to the fixative to a final concentration of 0.1% (w/v). Tissues were washed four times in PBS and incubated overnight at 4°C with primary antibodies, washed again with PBS, and incubated for 2 h with secondary antibodies. Samples were mounted in VectaShield before imaging. The antibodies used were rabbit anti-pAkt-505 (1:100; Cell Signaling), mouse anti-Dl (1:30; Developmental Studies Hybridoma Bank), Alexa 555 anti-mouse, Alexa 647 anti-mouse, and Alexa 488 anti-rabbit IgG. For proliferation assays, midguts were incubated in 5 µM EdU for 30 min immediately after dissection according to the manufacturer’s instructions (Invitrogen). Nuclei were visualized with DAPI.
3
Drosophila Leg Disc Staining and Quantification
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Larval and prepupal leg discs were fixed and stained following standard procedures. As primary antibodies we used rabbit and mouse anti-βGal, rabbit anti-Dys (a gift from L. Jiang and S.T. Crews), rabbit anti-DCas-3 (cleaved Drosophila Dcp-1, Cell Signaling Technology), rabbit anti-P-Mad (kindly provided by G. Morata). Mouse anti-Dl, anti-Dlg and anti-Tgo are from Developmental Studies Hybridoma Bank, University of Iowa. TRITC-phalloidin and Phalloidin-Atto 647N were used to stain F-actin (Sigma Aldrich), and secondary antibodies were coupled to Red-X, FITC and Cy5 fluorocromes (Alexa Fluor Dyes, Invitrogen).
To determine the levels of cell death in E(spl)-mβ and “fold” domains, we have performed Z-stack imaging of wild type (n = 8 prepupae leg discs) and dys2/dys3 mutants (n = 10 prepupae leg discs) and counted the number of D-Cas3 positive cells on each domain with the aid of the Fiji software. We selected for this analysis the joints between tarsal segments 2/3 and 3/4.
To determine the levels of cell death in E(spl)-mβ and “fold” domains, we have performed Z-stack imaging of wild type (n = 8 prepupae leg discs) and dys2/dys3 mutants (n = 10 prepupae leg discs) and counted the number of D-Cas3 positive cells on each domain with the aid of the Fiji software. We selected for this analysis the joints between tarsal segments 2/3 and 3/4.
4
Immunostaining and FISH Protocols
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Immunostaining and FISH protocols were followed as previously described (Kosman et al. 2004 (link)). Sheep antidigoxigenin (Life Technology PA185378), or rabbit anti-FITC (Invitrogen A889), mouse anti-Dl (1:10; Developmental Studies Hybridoma Bank 7A4) or guinea pig anti-Twi (1:200) (Trisnadi and Stathopoulos 2015 (link)) were used together with Alexa conjugate secondaries (1:400; Thermo Fisher). DAPI staining (1:10,000; Molecular Probes) was used to mark nuclei.
5
Immunofluorescent Staining Protocol
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Immunofluorescent staining was carried out using a standard procedure (König and Shcherbata, 2013 (link)). The following primary antibodies were used: mouse anti-En (1:20), mouse anti-β-Gal (1:20), mouse anti-BrZ1 (1:20), mouse anti-NICD (1:20) and mouse anti-Dl (1:20), all from Developmental Studies Hybridoma Bank; rabbit anti-phosphorylated Mad (pMad, 1:5000; a gift from Ed Laufer, Columbia University, New York, USA), rabbit anti-Vasa (1:5000; a gift from Herbert Jäckle, Max Planck Institute for Biophysical Chemistry, Goettingen, Germany), mouse anti-CD2 (1:100, BioLegend, 100107), guinea pig anti-Traffic Jam (Tj, 1:5000; a gift from D. Godt, University of Toronto, Canada) and chicken anti-GFP (1:5000, Abcam, ab13970). Secondary antibodies were: goat anti-rabbit Alexa 488, goat anti-chicken Alexa 488 and goat anti-guinea pig Alexa 647 (1:500, Life Technologies, A-11034, A-11039, A-21450); goat anti-mouse Cy3 IgG1 and goat anti-mouse IgG2a Alexa 488 (1:250, Jackson ImmunoResearch Laboratory). For visualizing cell nuclei, DAPI dye was used (Sigma). Samples were analyzed using a confocal microscope (Zeiss LSM 700). For making figures, Adobe Photoshop software was used.
6
Immunofluorescence Antibody Panel
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The following primary antibodies diluted in 1× PBST were used in this study: mouse anti-Dl, mouse anti-Pros, mouse anti-HP1 (Developmental Studies Hybridoma Bank, Iowa City, IA, USA), 1:200; mouse anti-GFP and rabbit anti-GFP (Molecular Probes,), 1:1000; rat anti-GFP (Nacalai Tesque Inc., Kyoto, Japan), 1:1000; rabbit anti-phospho-histone H3 (PH3; Millipore, Billerica, MA, USA), 1:1000; anti-Cleaved caspase-3 (Cell Signaling Technologies, Danvers, MA, USA), 1:1000; rabbit anti-γH2AvD (Rockland, Gilbertsville, PA, USA), 1:2000; mouse anti-γ-tubulin (Sigma-Aldrich), 1:1000; and anti-hVDR antibody (Thermo Fisher Scientific, Cleveland, OH, USA), 1:1000. The following secondary antibodies diluted in 1× PBST were used in this study: goat anti-rabbit FITC; goat anti-rabbit Cy3; goat anti-mouse FITC, goat anti-mouse Cy3, goat anti-rat FITC, and goat anti-rabbit Alexa Fluor® 647 (Jackson ImmunoResearch, West Grove, PA, USA), 1:400.
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