Dissected tissues were fixed 5 to 20 minutes in 4% formaldehyde/PBS depending on the primary antibody. Stainings were performed in 0.5% Triton/PBS with antibody incubations separated by several washes. Tissues were then transferred in Vectashield with or without DAPI for image acquisition. Primary antibodies were chicken anti-GFP (1:1,000, Aves #GFP-1020), rabbit anti-RFP (1:500, Rockland #600-401-379), rat anti-RFP (1:500, Chromotek #5F8), mouse anti-Cut (1:50, Developmental Studies Hybridoma Bank [DSHB] #2B10), mouse anti-Br-core (1:50, DSHB #25E9.D7), mouse anti-Br-Z1 (1:50, DSHB #Z1.3C11.OA1), mouse anti-Br-Z3 (1:50, DSHB #Z3.9A7), rabbit anti-Br-Z2 (1:50, Y. Song), mouse anti-Mmp1 (1:100, a combination of DSHB #14A3D2, 3A6B4, and 5H7B11), mouse anti-EcR (1:7, DSHB #Ag10.2), rabbit anti-ß-galactosidase (1:1,000, Cappel #559562), guinea pig anti-Sens (1:1,000, H. Bellen), mouse anti-Wg (1:100, DSHB #4D4), rabbit anti-Pdm1/Nub (1:500, S. Cohen), rat anti-Chinmo (1:500, N. Sokol), and guinea pig anti-Chinmo (1:500, N. Sokol). Rabbit anti-cleaved Dcp-1 (1:500, Cell Signaling #9578) labels apoptotic cells. Adequate combinations of secondary antibodies (Jackson ImmunoResearch) were used to reveal expression patterns.
Gfp 1020
Manufactured by Rockland Immunochemicals
Sourced in United States
The GFP-1020 is a fluorescent protein that emits green light when excited by a specific wavelength of light. It is commonly used as a marker in biological research to track the expression and localization of proteins of interest.
Automatically generated - may contain errors
Lab products found in correlation
Fluoromount g, by Southern Biotech (2 mentions)
Anti ha, by Abcam (2 mentions)
Anti cleaved caspase 3 asp175, by Cell Signaling Technology (2 mentions)
Anti actin, by Abcam (2 mentions)
Anti rad51, by Developmental Studies Hybridoma Bank (2 mentions)
Anti lamin dm0, by Developmental Studies Hybridoma Bank (2 mentions)
Anti γh2av, by Rockland Immunochemicals (2 mentions)
Anti gfp, by Thermo Fisher Scientific (2 mentions)
Anti h3k9me3, by Active Motif (2 mentions)
Anti flag, by Merck Group (2 mentions)
Mab414, by Fortrea (2 mentions)
Mab377, by Merck Group (2 mentions)
Alexa 555, by Thermo Fisher Scientific (1 mentions)
Alexa 647, by Thermo Fisher Scientific (1 mentions)
U4757, by Merck Group (1 mentions)
Alexa 488, by Thermo Fisher Scientific (1 mentions)
A 21235, by Thermo Fisher Scientific (1 mentions)
A21428, by Thermo Fisher Scientific (1 mentions)
Superfrost gold plus microscopy slides, by Thermo Fisher Scientific (1 mentions)
Tcs sp5 confocal microscope, by Leica (1 mentions)
10 protocols using gfp 1020
1
Immunohistochemical Staining of Drosophila Tissues
2
Immunohistochemical Analysis of Neuropeptides
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The following primary antibodies were used: rabbit anti-Ucn1 at 1/2000th (Sigma U4757), goat anti-CART at 1/2000th (R&D systems AF163), chicken anti-GFP at 1/3000th (Aves GFP-1020), Goat anti-RFP at 1/1000th (Rockland 200-101-379), rabbit anti-PAQR8/mPRβ at 1/1000th (Bioss USA, BS11410R), rabbit anti-PAQR5/mPRγ at 1/1000th (Abcam ab236798), rabbit anti-Fos at 1/3000th (Synaptic Systems 226-003), guinea pig anti-Fos at 1/1000th (Synaptic Systems 226-004), mouse anti-NeuN at 1/1000th (Millipore MAB377), rabbit anti-Progesterone Receptor at 1/500th (Sigma SAB4502184).
Secondary antibodies raised in donkeys and coupled with either Alexa488, Alexa555 or Alexa647 were sources from Thermo Fisher Scientific and used at 1/1000th.
Secondary antibodies raised in donkeys and coupled with either Alexa488, Alexa555 or Alexa647 were sources from Thermo Fisher Scientific and used at 1/1000th.
3
Immunohistochemistry of Brain Tissues
4
Tissue Fixation and Immunohistochemistry Protocol
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Animals were transcardially perfused with PBS followed by 4% paraformaldehyde. Brains were dissected and post-fixed for 16–24 h at 4 °C. Brains were sectioned using a vibratome (Leica) to a thickness of 50 µm. Samples were then stained with the following primary antibodies: anti-GFP (Aves Labs GFP-1020) at 1:1,000, anti-mCherry (Rockland 600-401-379) at 1:1,000 or anti-FOS antibody (Synaptic Systems 226 003) at 1:2,000 in PTxwH buffer for 24 h at 4 °C. After four 15-min washes in PTxwH buffer, samples were incubated in Alexa Fluor-conjugated secondary antibodies (Thermo Fisher) at 1:2,000 for 90 min at room temperature, followed by four further 15-min washes before mounting using 4′,6-diamidino-2-phenylindole Fluoromount-G (SouthernBiotech 0100-20 OB010020).
5
Multicolor Immunofluorescence in Barrel Cortex
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Barrel cortex sections were washed in TRIS buffer (TB) 1 × 15 min, TRIS-buffered saline (TBS) 1 × 15 min, and TBS + 0.5% Triton X-100 (TBST) 2 × 15 min, all at pH 7.6. For blocking, sections were incubated 90 min at room temperature in 0.25% bovine serum albumin/10% goat serum/TBST (Jackson ImmunoResearch). For primary antibody labeling, sections were incubated 48–72 h at 4 °C with (1) chicken anti-GFP (Aves; Cat#GFP-1020, RRID:AB_10000240) diluted 1:500 and (2) mouse anti-RFP (Rockland; Cat#200–301-379S, RRID:AB_2611064) diluted 1:2000. Sections were washed 4 × 15 min with TBST. For secondary antibody labeling, sections were incubated 4 h at room temperature with (1) Alexa Fluor 488-conjugated goat anti-chicken IgG (Molecular Probes; Cat#A11039) and (2) Alexa Fluor 568-conjugated goat anti-mouse IgG2a (Molecular Probes; Cat#A21134) diluted 1:500 in TBST. In some cases, sections were stained in addition with the primary antibody guinea pig anti-vGluT2 (Millipore; Cat#AB2251, RRID: AB_1587626) diluted 1:2000 and the secondary antibody Alexa Fluor 633-conjugated goat anti-guinea pig (Molecular probes; Cat#A21105) diluted 1:500. After washing 2 × 15 min with TBST and 1 × 15 min with TBS, sections were stained with DAPI, diluted 1:1000 in TBS. After several washes in TB, sections were mounted in Aqua-Poly-Mount (Polysciences).
6
Comprehensive Antibody Resource for Research
7
Comprehensive Antibody Resource for Research
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Antibodies used were: anti-Rad51 (1:1000, gift from J. Kadonaga); anti-Lamin/Dm0 (1:500, Developmental Studies Hybridoma Bank, ADL101); anti-Actin (1:1000, Abcam, ab8224); anti-HA (1:1000, Abcam, ab9134 for Western blot; 1:1000, Covance, 16B12 for IF); anti-FLAG (1:1000, Sigma, F1804); anti-Koi (1:1000, gift from J. Fischer and M. Welte); anti-Nup153 (1:200, SDI, Karpen Lab); anti-dPIAS (1:100, Karpen lab); anti-γH2Av (1:1000, Rockland, 600-401-914; 1:100 Developmental Studies Hybridoma Bank, UNC93-5.2.1); anti-GFP (1:1000, Invitrogen, AP11122 for Western blot; 1:1000 Aveles Lab, GFP-1020 for IF; Rockland, 600-101-215 for Ip); anti-Nup107 (1:2000, gift from V. Doye); anti-H3K9me2 (1:500, Upstate, 07-442; 1:750, Wako Chemicals, MABI0307, 302-32369); anti-H3K9me3 (1:500, Active Motif, 39161); anti-SUMO (1:500, SDI, Karpen Lab); anti-Mtor (1:50, SDI, Karpen Lab); anti-FG porins80 (link) (1:100, Covance, MAb414, MMS-120R); anti-cleaved Caspase3/Asp175 (1:500, Cell Signaling, 5A1E, #9664). Secondary antibodies for IF were from Life Technologies and Jackson Immunoresearch. Those used for western blot were from Pierce and Santa Cruz Biotech.
8
Amplifying Fluorescent Signals in Brain Samples
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Samples were washed twice in PTx.2 at RT, followed by permeabilization for 2 days at 37 °C. Samples were again washed twice in PTx.2 at RT, then blocked for 3 days at 37 °C. To amplify the host AAV1-GCaMP6s and transplanted PV-tdTomato signals, samples were washed twice in PTwH at RT and incubated for 6 days at 37 °C with 1 : 500 chicken anti-GFP (Aves Labs, GFP-1020) and 1 : 400 rabbit anti-RFP (Rockland, 600-401-379), respectively. Samples were then washed in PTwH for increasing durations of 0 min to 2 h for the first day, 2 h each for the second. Following a secondary incubation for 6 days at 37 °C with 1 : 500 donkey anti-chicken Alexa 647 (Jackson ImmunoResearch, 703-605-155) and 1 : 400 donkey anti-rabbit Alexa 568 (Abcam, ab175692), samples were washed using the same post-primary protocol.
9
Immunostaining for Neural Cell Markers
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Sections without signs of tissue damage were chosen for immunostaining (see also Figure S1 A). The immunohistology protocol was followed as previously described (Bao et al., 2017 (link)). The following primary antibodies were used: mouse anti-GFAP (Millipore #MAB360, 1:1,000), mouse anti-NeuN (Millipore #MAB377, 1:1,000), rabbit anti-GABA (Sigma #A2052, 1:500), goat anti-MCM2 (R&D Systems #AF5778, 1:500), chicken anti-Nestin (Aves Labs #NES, 1:1,000), chicken anti-GFP (Aves Labs #GFP-1020, 1:1,000), rabbit anti-RFP (Rockland #600-401-379, 1:1,000), rabbit anti-S100B (Abcam #ab52642, 1:1,000), rat anti-mCherry (Invitrogen #M11217, 1:500), goat anti-DCX (Santa Cruz Biotechnology #sc-8066, 1:100), goat anti-GFP (Rockland #600-101-215, 1:500), and goat anti-SOX2 (Santa Cruz #sc-17320, 1:250). All fluorescent secondary antibodies were obtained from Life Technologies and diluted to 1:500.
All images for quantification were acquired as a z stack, ≤1 μm axial step, then loaded into ImageJ (Fiji) software as a composite image and counted using the Cell Counter plugin. For 3D reconstruction, images were deconvolved using Huygens Professional software (SVI, the Netherlands). 3D reconstruction data were displayed using Surface Renderer in Huygens.
All images for quantification were acquired as a z stack, ≤1 μm axial step, then loaded into ImageJ (Fiji) software as a composite image and counted using the Cell Counter plugin. For 3D reconstruction, images were deconvolved using Huygens Professional software (SVI, the Netherlands). 3D reconstruction data were displayed using Surface Renderer in Huygens.
10
Immunostaining of Mouse Cortical Tissue
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Tissues were processed as per mouse cortical tissue. Before sections were blocked they were post-fixed with 4% PFA for 10 mins, permeabilized in 0.3% Triton-X100 and then blocked with 0.1% Tween, 3% BSA and 10% normal goat serum. Sections then incubated with primary antibodies diluted in blocking solution. GFP (chick, Aves Lab, Cat. # GFP-1020, 1:1000) and RFP (rbb, Rockland 1:1000, Cat. # 600-401-379).
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