Immunoprecipitation of nuclear extracts from an overnight collection of embryos was performed following the RNA immunoprecipitation protocol of Hilgers et al. (Hilgers et al., 2012 (link)), except that the nuclear extracts were incubated overnight with 2 μg mouse anti-ELAV 9F8A9 or mouse anti-Tub E7 antibodies (Developmental Studies Hybridoma Bank).
Mouse anti elav 9f8a9
Manufactured by Developmental Studies Hybridoma Bank
Mouse anti-ELAV 9F8A9 is a monoclonal antibody that recognizes the ELAV (Embryonic Lethal Abnormal Vision) protein. The ELAV protein is an RNA-binding protein that plays a role in the regulation of gene expression. The 9F8A9 clone of the mouse anti-ELAV antibody is a useful tool for the detection and study of the ELAV protein in various experimental systems.
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Lab products found in correlation
Fitc conjugated goat anti rabbit igg, by Jackson ImmunoResearch (1 mentions)
Cy3 conjugated goat anti mouse igg, by Jackson ImmunoResearch (1 mentions)
Mouse anti repo 8d12, by Developmental Studies Hybridoma Bank (1 mentions)
Vectashield, by Vector Laboratories (1 mentions)
Sab4502380, by Merck Group (1 mentions)
Alexa fluor 647 conjugated phalloidin, by Thermo Fisher Scientific (1 mentions)
Mouse anti armadillo n2 7a1, by Developmental Studies Hybridoma Bank (1 mentions)
Mouse anti myc 9b11, by Cell Signaling Technology (1 mentions)
Mouse anti ha 6e2, by Cell Signaling Technology (1 mentions)
Tcs sp5, by Leica (1 mentions)
Anti rat rhodamine, by Jackson ImmunoResearch (1 mentions)
A6455, by Thermo Fisher Scientific (1 mentions)
Anti guinea pig cy5, by Jackson ImmunoResearch (1 mentions)
4 protocols using mouse anti elav 9f8a9
1
Immunoprecipitation of Embryonic Nuclear Extracts
2
Immunostaining of Drosophila Nervous System
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For the dSap-r expression pattern, third instar larvae were dissected and fixed in 3.7% formaldehyde/PBS for 10 min, followed by 3 × 5 min washes in 0.1% PBST (PBS with 0.1% Triton X-100). Larvae were labelled overnight at 4 °C with mouse anti-repo-8D12 or mouse anti-elav-9F8A9 diluted in PBST (1:50; Developmental Studies Hybridoma Bank, University of Iowa). Washes were performed as above, followed by incubation for 2 h at RT in Cy3-conjugated goat anti-mouse IgG (1:200; Jackson ImmunoResearch). Larvae were washed (as above) and left in 70% glycerol/PBS for 1–2 h before mounting in Vectashield (Vector Laboratories). To label lysosomes, aged adult brains were dissected in 4% paraformaldehyde/PBS and transferred to fresh fixative for 20 min. Brains were washed 3 × 15–20 min in 0.3% PBST followed by incubation overnight at RT with rabbit anti-Arl-8 (1:500; kindly provided by Debbie Smith, University of York, U·K) and mouse anti-elav (1:50). Brains were washed as above, followed by incubation for 3 h at RT in FITC-conjugated goat anti-rabbit IgG and Cy3-conjugated goat anti-mouse IgG (1:200; Jackson ImmunoResearch). Brains were washed and mounted in Vectashield. All images were acquired using a Zeiss LSM 510 meta Axiovert 200M laser scanning confocal microscope.
3
Immunofluorescence and TEM Imaging Protocols
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All procedures (TEM and Immunofluoresence) were performed as previously described [14 (link), 15 (link)]. The primary antibodies used in this study were: mouse anti-elav (9F8A9, 1:200, Developmental Studies Hybridoma Bank); mouse anti-Crumbs (Cq4, 1:100, Developmental Studies Hybridoma Bank), rabbit anti-aPKC ζ (zeta) (1:200, SAB4502380 Sigma), mouse anti-HA (6E2, 1:500, Cell Signaling), mouse anti-myc (9B11, 1:500 Cell Signaling), rat anti-Crumbs (1:500) [73 (link)], mouse anti-armadillo (N2 7A1, 1:100, Developmental Studies Hybridoma Bank). The C-terminal peptide KVNKLISRFEGGRPRLC (produced in the laboratory of Dr. Charles Zuker) was used as the antigen in rabbits, and the resulting antibody was used at a concentration of 1:200. Fluorescent conjugated secondary antibodies were obtained from either Jackson ImmunoResearch Laboratories or Life Technologies. Rhodamine or Alexa Fluor 647 conjugated phalloidin (1:200, Life Technologies) were utilized for the detection of F-Actin. Confocal images were captured on a Leica TCS SP5. TEM imaging was conducted with a JOEL 1010 and JOEL 1400. All images were processed in Adobe Photoshop.
4
Antibody Immunostaining Protocol for Drosophila
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Antibody stains were performed following standard procedures. Primary antibodies were monoclonal mouse anti-Ubx (FP3.38, a gift from Robert White, University of Cambridge; 1:20), mouse anti-Antp (4C3; 1:20), mouse anti-Abd-B (1A2E9; 1:20), rat anti-ELAV (7E8410; 1:300) and mouse anti-ELAV (9F8A9; 1:300) (all from Developmental Studies Hybridoma Bank); goat anti-Abd-A (dH-17, Santa Cruz Biotechnology; 1:20); rabbit anti-Eg (Dittrich et al., 1997 (link); 1:500); mouse anti-Eg (a gift from Chris Doe, University of Oregon, USA; 1:100); rabbit cleaved Drosophila Dcp-1 (Asp216, Cell Signaling; 1:50); guinea pig anti-Hb (J. Urban, University of Mainz, Germany; 1:500), rabbit anti-β-gal (A11132, Molecular Probes; 1:300), rabbit anti-GFP (A6455, Molecular Probes; 1:300) and rabbit anti-Repo (as described by Halter et al., 1995; 1:500). Secondary antibodies used were anti-mouse-A488 (A21202; 1:500), anti-rat-A488 (A21202; 1:500) and anti-rabbit-Alexa568 (A10042; 1:500) (all from Molecular Probes); anti-rabbit-Rhodamine (711-025-152; 1:500), anti-rat-Rhodamine (712-026-153; 1:500), anti-guinea pig-Cy5 (706-175-148: 1:500), anti-rat-DyLight 405 (712-475-153; 1:500), anti-goat-Cy3 (705-165-003; 1:500) (all from Jackson ImmunoResearch Laboratories).
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