Rat anti deadpan

Manufactured by Abcam

Rat anti-Deadpan is a primary antibody that recognizes the Deadpan protein in rats. Deadpan is a transcription factor that plays a role in neural development.

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Lab products found in correlation

Goat anti rabbit cy3, by Jackson ImmunoResearch (1 mentions) Goat anti rat alexa 488, by Jackson ImmunoResearch (1 mentions) Mouse anti fasciclin 2, by Developmental Studies Hybridoma Bank (1 mentions) Goat anti mouse alexa 488, by Jackson ImmunoResearch (1 mentions) Rabbit anti phospho histone h3 ph3, by Cell Signaling Technology (1 mentions) Alexa fluor dye, by Thermo Fisher Scientific (1 mentions) Click it edu imaging kit, by Thermo Fisher Scientific (1 mentions) Chicken anti gfp, by Abcam (1 mentions) Mouse anti repo 8d12, by Developmental Studies Hybridoma Bank (1 mentions) Ab195172, by Abcam (1 mentions) Mouse anti prospero mr1a, by Developmental Studies Hybridoma Bank (1 mentions) Mouse anti strep, by Qiagen (1 mentions) Rat anti elav 7e8a10, by Developmental Studies Hybridoma Bank (1 mentions) Lsm880 confocal microscope, by Adobe (1 mentions) Donkey secondary antibodies, by Jackson ImmunoResearch (1 mentions) Mouse anti ha, by Fortrea (1 mentions) Mouse anti prospero, by Developmental Studies Hybridoma Bank (1 mentions)

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Anti-Deadpan (3 citations)

5 protocols using rat anti deadpan

Immunodetection of Neural Progenitors

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Antibodies for neuroblast (NB) and intermediate neural progenitor (INP) detection were used at the following dilutions in 0.3% PBST (PBS + 10% Triton-X 100): Rabbit anti-phospho-Histone H3 (pH3; Cell Signaling) at 1:500, rat anti-Deadpan (Abcam) at 1:75, goat anti-rabbit Cy3 (Jackson Immunoresearch) at 1:300, and goat anti-rat Alexa 488 (Jackson ImmunoResearch) at 1:250. Anti-Deadpan marks all NBs and INPs and was used to define and exclude the optic lobe during analysis. Anti-pH3 marks all proliferating NBs and progenitor cells in M phase. Antibodies used for the detection of fasciculating axons in the mushroom body were used at the following dilutions in 0.1% PBST: Mouse anti-FasciclinII (Developmental Studies Hybridoma Bank) at 1:20 and goat anti-mouse Alexa 488 (Jackson ImmunoResearch) at 1:1000.

Nguyen U., Tinsley B., Sen Y., Stein J., Palacios Y., Ceballos A., Welch C., Nzenkue K., Penn A., Murphy L., Leodones K., Casiquin J., Ivory I., Ghenta K., Danziger K., Widman E., Newman J., Triplehorn M., Hindi Z, & Mulligan K. (2020). Exposure to bisphenol A differentially impacts neurodevelopment and behavior in Drosophila melanogaster from distinct genetic backgrounds. Neurotoxicology, 82, 146-157.

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Larval Brain Dissection and Staining

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Larval brains were dissected in PBS and fixed in 4% paraformaldehyde/PEM for 20 min, followed by detection of EdU using the Click-iT EdU imaging kit (Molecular Probes, Eugene, OR) (Doyle et al., 2017 (link)). After thorough washing in 0.1% Triton-x/PBS, antibody staining was performed according to standard methods (Doyle et al., 2017 (link); Siegrist et al., 2010 (link)). Primary antibodies used in this study were: chicken anti-GFP (1:100, Abcam, Cambridge, MA), rat anti-Deadpan (1:100, Abcam), rabbit anti-Scribble (1:200; gift of C. Doe), and rabbit anti-Eyeless (1:1000; gift of U. Waldorf). Secondary antibodies were conjugated to Alexa Fluor dyes (Molecular Probes).

Sipe C.W, & Siegrist S.E. (2017). Eyeless uncouples mushroom body neuroblast proliferation from dietary amino acids in Drosophila. eLife, 6, e26343.

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Quantification of Brain Lobe Volume

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Brains were dissected in PBS and fixed with 4% PFA/PBS/0.3%Triton for 20 minutes and blocked in PBS/0.3%Triton/1%BSA/5% normal goat serum and incubated in primary antibody in PBS/0.3%Triton/1%BSA overnight. Primary antibodies include guinea pig anti-Deadpan (gift from Chris Doe, originally from Jim Skeath, 1:1000), rat anti-Deadpan (Abcam ab195172, 1:250 or 1:500), mouse anti-Prospero MR1A (Developmental Studies Hybridoma Bank, 1:1000), rat anti-Elav 7E8A10 (Developmental Studies Hybridoma Bank, 1:250) and mouse anti-Repo 8D12 (Developmental Studies Hybridoma Bank, 1:250) and mouse anti-Strep (Qiagen, 1:500) with goat secondary antibodies from Jackson Laboratories used 1:500. Brains were mounted with tape spacers and imaged using a Leica Sp8 with 2 βm sections through the entire brain lobe. Resulting stacks were analyzed using the Surfaces function in Imaris (Bitplane) to quantify brain lobe volume. One lobe from each brain was analyzed.

Shah P.S., Link N., Jang G.M., Sharp P.P., Zhu T., Swaney D.L., Johnson J.R., Von Dollen J., Ramage H.R., Satkamp L., Newton B., Aguirre S., Hüttenhain R., Petit M.J., Baum T., Everitt A., Laufman O., Tassetto M., Shales M., Stevenson E., Iglesias G.N., Shokat L., Tripathi S., Balasubramaniam V., Webb L.G., Willsey A.J., Garcia-Sastre A., Pollard K.S., Cherry S., Gamarnik A.V., Marazzi I., Taunton J., Fernandez-Sesma A., Bellen H.J., Andino R, & Krogan N.J. (2018). Comparative flavivirus-host protein interaction mapping reveals mechanisms of dengue and Zika virus pathogenesis. Cell, 175(7), 1931-1945.e18.

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Immunostaining of Drosophila Nervous System

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Fly brains at indicated larval, pupal, and adult stages were dissected, fixed and immunostained as described previously (37 (link),39 (link)). The following primary antibodies were used in this study: chicken GFP polyclonal antibody (1:500, Invitrogen, A10262); rat anti-Deadpan (1:100, abcam, ab195173); mouse 1D4 anti-Fasciclin II (1:50, DSHB); rabbit polyclonal anti-Trio (1:1000) (46 (link)); rat anti-Chinmo (1:500, a gift from the Sokol lab) (47 (link)). Secondary antibodies from Molecular Probes were all used in a 1:200 dilution. The immunofluorescent signals were collected using Zeiss LSM 880 confocal microscope and processed using Fiji and Adobe Illustrator.

Chen H.M., Marques J.G., Sugino K., Wei D., Miyares R.L, & Lee T. (2020). CAMIO: a transgenic CRISPR pipeline to create diverse targeted genome deletions in Drosophila. Nucleic Acids Research, 48(8), 4344-4356.

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Late Third Instar Larval Brain Immunostaining

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Late third instar (judged based on gut clearance and extruding spiracles) larval brains were dissected in PBS and fixed with 4% PFA/PBS/0.3% Triton for 20 minutes. For immunostaining, brains were blocked in PBS/0.3% Triton/1% BSA/5% normal goat serum and incubated in primary antibody in PBS/0.3% Triton/1% BSA overnight. Primary antibodies include rat anti-Deadpan (Abcam Cat# ab195172, 1:250 or 1:500), mouse anti-Prospero (Developmental Studies Hybridoma Bank, MR1A, 1:1000), and mouse anti-HA (Covance Cat# 901501, 1:1000) with donkey secondary antibodies from Jackson ImmunoResearch used at 1:500. Brains were mounted with double sided tape spacers and imaged using a confocal microscope (Leica Sp8) with 2 μm or 3 μm sections through the entire brain lobe.

Link N., Harnish J.M., Hull B., Gibson S., Dietze M., Mgbike U.E., Medina-Balcazar S., Shah P.S, & Yamamoto S. (2023). A Zika virus protein expression screen in Drosophila to investigate targeted host pathways during development. bioRxiv.

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