Chicken-anti-βgal (1:200, Immunology Consultants Laboratory), rabbit-anti-βgal (1:1000, Molecular Probes), mouse-anti-CycB (1:100, Developmental Studies Hybridoma Bank/DSHB), rat-anti-DEcad (1:50, DSHB), rabbit-anti-Dgo (1:100, Feiguin et al., 2001 (link)), rat-anti-Elav (1:50, DSHB), Mouse-anti-Fmi (1:10, DSHB), mouse-anti-GFP (1:1000, Roche), rabbit-anti-Nek2 (1:1000, Prigent et al., 2005 (link)), rat-anti-Pk (1:25; Strutt et al., 2013 (link)) mouse-anti-Ro (1:10, DSHB), mouse-anti-γTub (1:1000, Sigma), Rhodamine phalloidin (1:1000, Invitrogen), phalloidin-AF647 (1:1000, Jackson IRL), Hoechst 33342 (10mg/ml, 1:500). All fluorophore coupled secondaries were from Jackson IRL used at 1:200.
Rabbit anti β gal
Manufactured by Thermo Fisher Scientific
Rabbit anti-β-gal is a primary antibody produced in rabbits that specifically binds to the β-galactosidase (β-gal) protein. It can be used in various laboratory applications that involve the detection and quantification of β-gal expression.
Automatically generated - may contain errors
Lab products found in correlation
Mouse anti 22c10, by Developmental Studies Hybridoma Bank (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions)
Lsm 700, by Zeiss (2 mentions)
Rabbit anti yki, by Thermo Fisher Scientific (2 mentions)
Mouse anti diap1, by Thermo Fisher Scientific (2 mentions)
Fluoview 1000 confocal microscope, by Adobe (2 mentions)
Rabbit anti dmyc, by Santa Cruz Biotechnology (2 mentions)
Anti rabbit igg conjugated to cy3, by Jackson ImmunoResearch (2 mentions)
Rhodamine phalloidin, by Thermo Fisher Scientific (1 mentions)
Mouse anti gfp, by Roche (1 mentions)
Mouse anti cycb, by Developmental Studies Hybridoma Bank (1 mentions)
Mouse anti γtub, by Merck Group (1 mentions)
Rabbit anti gfp, by Thermo Fisher Scientific (1 mentions)
Rabbit anti gfp, by Abcam (1 mentions)
Dab substrate kit, by Vector Laboratories (1 mentions)
Sheep anti digoxigenin ap, by Roche (1 mentions)
Mouse anti brdu g3g4, by Developmental Studies Hybridoma Bank (1 mentions)
Rabbit anti caspase 3, by Abcam (1 mentions)
Ab47651, by Abcam (1 mentions)
Goat anti chicken alexa 488, by Thermo Fisher Scientific (1 mentions)
10 protocols using rabbit anti β gal
1
Immunofluorescence Labeling of Drosophila Tissues
2
Drosophila Immunohistochemistry Protocol
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HRP and fluorescence immunohistochemistry were performed as described previously [26 (link)–28 (link)]. The following primary antibodies were used: rabbit anti-β-gal (1:3000, Molecular Probes), mouse anti-22C10 (1:10, Developmental Studies Hybridoma Bank (DSHB), University of Iowa, IA), rat anti-Elav (7E8A10) (1:10, DSHB), mouse anti-Repo (1:100, DSHB), mouse anti-Prospero (1:10, DSHB), mouse anti-Slit (C555.6D) (1:10, DSHB), mouse anti-Robo (13C9) (1:10, DSHB), rabbit anti-α-tubulin 85E (1:50, Thermo Fisher Scientific), and rabbit anti-GFP (1:5000, ThermoFisher Scientific). The following secondary antibodies were used at 1:500: anti-Mouse 555, anti-Rabbit 488, anti-Rat 488, anti-Rat 647, biotinylated anti-mouse and biotinylated anti-rabbit.
3
Immunohistochemical Visualization of β-Gal
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Following cryo-sectioning, immunohistochemistry was performed on P2-lacZ sections to visualize β-gal signal. Sections were equilibrated in 1× PBS for 5 min at room temperature (RT), fixed in 4% PFA in 1× PBS for 10 min at RT, and washed in 1× PBS for 2 × 3 min at RT. Sections were then incubated in 0.5% Triton-X-100 in 1× PBS for 30 min at RT, washed in 1× PBS for 3 × 5 min at RT, and incubated in 5% normal goat serum in 1× PBS (blocking buffer) for 30 min at RT. Finally, sections were incubated in primary antibodies in blocking buffer overnight at 4 °C, washed in 1× PBS for 3 × 3 min at RT, and incubated in secondary antibodies in blocking buffer for 2 h at RT.
The primary antibody used was rabbit anti-β-gal (Molecular Probes) at a dilution of 1:500 in blocking buffer. The secondary antibody used was goat anti-rabbit conjugated Alexa Fluor 594 (Life Technologies) at a dilution of 1:500 in blocking buffer.
The primary antibody used was rabbit anti-β-gal (Molecular Probes) at a dilution of 1:500 in blocking buffer. The secondary antibody used was goat anti-rabbit conjugated Alexa Fluor 594 (Life Technologies) at a dilution of 1:500 in blocking buffer.
4
Immunofluorescence Analysis of Fixed Cardiac Tissue
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The harvested hearts were fixed with 4% paraformaldehyde and embedded in paraffin. The sections were then immunostained with the following primary antibodies: mouse anti-GFP (1:500; MBL), rabbit anti-GFP (1:200; Abcam or GeneTex), rabbit anti-β-Gal (1:500; Invitrogen), mouse anti-cTnT (1:100; DSHB), and rat anti-Sca-1-PE (1:500; BD Bioscience). A DAB substrate kit (Vector Laboratories) was used for immunohistochemistry and appropriate secondary antibodies (Invitrogen or Abcam) were used for visualization under a fluorescence microscope. The plasma membrane was immunostained with wheat germ agglutinin (WGA, 5 μg/ml, Invitrogen) and 4,6-diamidino-2-phenylindole (DAPI, 1 μg/ml; Sigma) was used for nucleus staining.
5
Immunostaining of Drosophila Wing Discs
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Third instar larvae were dissected in cold phosphate-buffered saline (PBS) and fixed in 4% formaldehyde/PBS for 20 min at room temperature. They were then washed and permeabilized in PBT (0.2% Triton X-100 in PBS) for 30 min and blocked in BBT (0.3% BSA, 250 mM NaCl in PBT) for 1 h. Samples were incubated overnight at 4°C with primary antibody diluted in BBT, washed three times (15 min each) in BBT and incubated with secondary antibodies and DAPI (1 μg/ml) for 1.5 hour at room temperature. After three washes with PBT (15 min each), wing discs were placed in mounting medium (80% glycerol/PBS containing 0.05% n-Propyl-Gallate). All steps were performed on a rocking platform at the indicated temperature. The following primary antibodies were used: mouse anti-BrdU (G3G4; Developmental Studies Hybridoma Bank (DSHB)); mouse anti-MMP1 (3A6B4, DSHB); mouse anti-p53 (7A4, DSHB); rabbit anti-p-Histone H3 (sc-8656, Santa Cruz); rabbit anti-β-Gal (A11132, Invitrogen); and sheep anti-Digoxigenin-AP (#11093274910, Roche). The following secondary antibodies were used: anti-mouse IgG-Alexa Fluor 594; anti-mouse IgG-Alexa Fluor 488; anti-rabbit IgG-Alexa Fluor 594; and anti-rabbit IgG-Alexa Fluor 488 (Jackson InmunoResearch).
6
Immunostaining and Microscopy Analysis
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Tissues were dissected and fixed in 4% formaldehyde. Antibody staining was performed as described before14 (link). The following antibodies were used: rabbit anti-DgC-term52 (link) (1:2,000), rabbit anti-Dgex8 (ref. 53 ) (1:500), mouse anti-22C10, anti-β-Gal, anti-Elav, anti-βPS Integrin, anti-Pcan, anti-Arm, anti-Pros and rat anti-DE-Cad (1:50; Developmental Studies Hybridoma Bank), chicken anti-GFP (1:2,000; Abcam), rabbit anti-β-Gal (1:1,000; Invitrogen), rabbit anti-Phospho-Histone 3 (anti-PH3, 1:2,000; Upstate Biotechnologies), guinea pig anti-Miranda (gift from Andreas Wodarz, 1:1,000), rabbit anti-Caspase3 (1:200; Abcam), rabbit anti-LanB (1:1,000, Abcam ab47651), rabbit anti-Dys186 (gift from Lee Fradkin), Alexa 488/568 goat anti-mouse, Alexa 488/568/633 goat anti-rabbit, Alexa 488 goat anti-mouse, Alexa 568 goat anti-guinea pig, Alexa 488 goat anti-chicken (1:500; Invitrogen). Samples were mounted on slides in 70% glycerol, 2% n-propyl gallate, 1 × PBS and analysed using a confocal microscope (Zeiss LSM 700). Quantification of staining intensities was done using the Zeiss software.
7
Immunostaining Analysis of Drosophila Hindgut
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Hindguts from experimental and control larvae, identified by the absence of the Tubby (Tb) marker, were dissected and processed as previously described (30 (link)). Dissections were performed in 1X PBS, dissected tissue fixed with 4% paraformaldehyde, and processed for immunohistochemistry using our standard protocols(28 (link), 30 (link)). The following primary antibodies were used: mouse anti-NimC1 (79 (link))(Gifted by Eva Kurucz, 1:30), mouse anti-Mmp1(1:1000, DSHB #3B8D12), mouse anti-phospho JNK (1:50, Cell Signaling Technology #9255), mouse anti-phospho Akt (1:1000, Cell Signaling Technology #4054), mouse anti-Histone 2A gamma variant, phosphorylated (1:100, DSHB #UNC93–5.2.1), rabbit anti-Histone H3 (tri methyl K9) (1:50, Abcam #ab8898), mouse anti-Dacapo (1:10, DSHB #NP1), rabbit anti-Dcp1(1:1000, Cell Signaling Technology #9578), mouse anti-β-gal (1:100, DSHB #40–1a), rabbit anti-β-gal (1:200, Thermo Fisher Scientific #A111–32 ). Alexa-conjugated goat-anti-mouse and anti-rabbit antibodies were used as secondary antibodies (1:1000, Thermo Fisher Scientific, #A-11031, #A-21052, #A-110356, #A-21071). All guts were imaged using SPE DM6 Leica Confocal Microscope at 40X magnification at 1.0 Zoom. Hindguts used to quantify the imaginal ring proliferation area were imaged at 10X magnification, 1.5X Zoom (30 (link)).
8
Hemocyte Isolation and Immunostaining in Drosophila
9
Immunohistochemical Analysis of Drosophila Tissues
10
Immunohistochemical Analysis of Drosophila Tissues
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All Drosophila stocks and crosses were maintained at 25°C.
Immunohistochemistry was done using a protocol described previously (Varelas et al.,
2008 ). The primary antibodies were rabbit anti-Yki (1:400; a gift from K. Irvine), mouse anti-DIAP1 (1:100; a gift
from B. Hay), rabbit anti-b-gal (1:100; Invitrogen), and rabbit anti-dMyc (1:100; Santa Cruz Biotechnology). The secondary
antibodies were donkey anti-mouse or anti-rabbit IgG conjugated to Cy3 (1:200; Jackson ImmunoResearch). Samples were imaged
using an Olympus Fluoview 1000 confocal microscope, and the images were edited using Adobe Photoshop CS6.0. Brain lobe size in
pixels was measured using the histogram function of Adobe Photoshop CS6.0. Data are presented as the means and standard
deviations calculated for each sample. A 2-tailed t test performed with Excel 2013 was used to determine whether the observed
differences were significant (p < 0.05).
Immunohistochemistry was done using a protocol described previously (
2008
from B. Hay), rabbit anti-b-gal (1:100; Invitrogen), and rabbit anti-dMyc (1:100; Santa Cruz Biotechnology). The secondary
antibodies were donkey anti-mouse or anti-rabbit IgG conjugated to Cy3 (1:200; Jackson ImmunoResearch). Samples were imaged
using an Olympus Fluoview 1000 confocal microscope, and the images were edited using Adobe Photoshop CS6.0. Brain lobe size in
pixels was measured using the histogram function of Adobe Photoshop CS6.0. Data are presented as the means and standard
deviations calculated for each sample. A 2-tailed t test performed with Excel 2013 was used to determine whether the observed
differences were significant (p < 0.05).
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