The following antibodies were used in this study: γ-tubulin (aa 38–53) GTU-88 (T6557, mouse, used at 1/10000 for WB and 1/1000 for IF/IHC, Sigma); Gamma-tubulin (434–451) TU-30 (ab27074, mouse, used at 1/1000, Abcam); GAPDH (MAB374, mouse, used at 1/1000, Chemicon); Cux1 (sc-13024, rabbit, used at 1/100, Santa Cruz); Cux1 (11733-AP, used at 1/100, Proteintech); GFP (A10262, chicken, used at 1/1000, ThermoFisher); CTIP2 (ab18465, rat, used at 1/500, Abcam); NeuN (MAB377, mouse, used at 1/500, Millipore); SATB2 (ab51502, mouse, used at 1/400, Abcam); TBR1 (ab31940, rabbit, used at 1/500, Abcam); Pax6 (PRB-278P, rabbit, used at 1/100, Eurogentec); Tbr2 (14–4875–80, rat, used at 1/200, eBioscience); PH3 (06–570, rabbit, used at 1/500, Millipore); Ki67 (NCL-L-Ki67-MM1, mouse, used at 1/500, Leica); Anti-tRFP (AB234, rabbit used at 1/5000 for WB, Evrogen); GCP4 (sc-271876, mouse, used at 1/1000 for WB, Santa Cruz); Anti-mouse antibody conjugated with HRP (W402B, goat, used at1/10000 for WB, Promega); Anti-rabbit antibody conjugated with HRP (W401B, goat, used at 1/100000 for WB, Promega). Anti-GCP2 antibody GCP2-01 (mouse monoclonal IgG2b) used for immunoprecipitation was described previously58 (link); Anti-GCP2 antibody GCP2-02 (mouse monoclonal IgG1, in the form of hybridoma spent culture supernatant, used at 1/10 for WB) was described previously58 (link).
Ab51502
Manufactured by Abcam
Sourced in United Kingdom
Ab51502 is a laboratory equipment product. It is a device used for scientific research and analysis purposes.
Automatically generated - may contain errors
Lab products found in correlation
Ab18465, by Abcam (15 mentions)
Ab31940, by Abcam (11 mentions)
Ab23345, by Abcam (6 mentions)
Mab377, by Merck Group (5 mentions)
Prb 278p, by Fortrea (4 mentions)
Sc 13024, by Abcam (3 mentions)
Lsm 710 confocal microscope, by Zeiss (3 mentions)
Ab6326, by Abcam (2 mentions)
Ab5392, by Abcam (2 mentions)
Ab79351, by Abcam (2 mentions)
Ab2253, by Merck Group (2 mentions)
Ab31940, by Santa Cruz Biotechnology (2 mentions)
A2228, by Merck Group (2 mentions)
Iblotting system, by Thermo Fisher Scientific (2 mentions)
Hpa029543, by Merck Group (2 mentions)
P8340, by Merck Group (2 mentions)
N ethylmaleimide, by Merck Group (2 mentions)
β mercaptoethanol, by Merck Group (2 mentions)
Laemmli sample buffer, by Bio-Rad (2 mentions)
Dc protein assay, by Bio-Rad (2 mentions)
36 protocols using ab51502
1
Antibody Characterization for Cell Biology Research
2
Immunohistochemical Labeling of Neural Markers
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Rat anti-BrdU (1:250, Abcam, #AB6326), rat anti-CTIP2 (1:500, Abcam, #AB18465), rabbit anti-CTIP2 (1:500, Abcam, #AB28448), rabbit anti-CUX1 (1:500, Santa Cruz, #sc-13024), rabbit anti-FITC (1:2000, Abcam, #AB19491), goat anti-FITC (1:1000, Novus Biolab, #NB600-493), mouse anti-SATB2 (1:200, Abcam, #AB51502), rabbit anti-TBR1 (1:500, Abcam, #AB31940).
3
Multiplexed Labeling of Brain Cells
4
Comprehensive Antibody Panel for Neuronal Characterization
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Rab10 (4262S, Cell Signaling Technology) for immunoblot, Rab10 (ab237703, Abcam) for immunofluorescence, pRab10 for immunoblot (ab230261, Abcam), pRab10 for immunofluorescence (ab241060, Abcam), Hsc70 (ab19136, Abcam), α-Tubulin (SC23948, Santa Cruz Biotechnology), parvalbumin, (NBP2-50036, NovusBio), NeuN (MAB377, Millipore), SATB2 (ab51502 Abcam), calretinin (MAB1568, Millipore), DARPP32 (MAB4230, R&D Systems), ChAT (NBP2-46620, NovusBio), DAT loop (6-8D6 Santa Cruz Biotechnology), tyrosine hydroxylase (TH) (ab76442, Abcam), CD68 (NBP2-33337SS, NovusBio), GFAP (AB5541, Millipore), Olig2 (MABN50 Millipore), KDEL receptor (sc-58774, Santa Cruz Biotechnology), TGN46 (MA3-063, ThermoFisher), LAMP1 (1D4B, DHSB), EEA1 (NBP2-36568, NovusBio), α-Synuclein (610786, BD Transduction Lab), Synuclein (ab51252, Abcam), VAMP2 (104 211, Synaptic Systems), Homer1 (160 006 Synaptic Systems), Rab8a (ab188574).
5
Immunoblotting and Immunostaining Protocols
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For immunoblotting, a polyclonal rabbit antibody (Calarco et al. 2009 (link)) raised against amino acids 1–82 of nSR100 was used at 1:5000. Anti-tubulin (T6074, Sigma) was used at 1:5000. For immunostaining, mouse monoclonal anti-neurofilament (2H3 conditioned medium, Iowa Developmental Studies Hybridoma Bank) was diluted to 1:50 for whole-mount diaphragm staining and 1:100 for brain section staining. Mouse anti-NeuN (mab377, Millipore), mouse anti-Satb2 (ab51502, Abcam), rabbit anti-Tbr1 (ab31940, Abcam), and chicken polyclonal anti-β-galactosidase (ab9361, Abcam) were all diluted to 1:500. Chicken anti-MAP2 (ab5392, Abcam) was diluted to 1:10,000, mouse anti-Tuj1 (MRB-435P, Covance) was diluted to 1:750, and rabbit anti-Pax6 (PRB-278P, Covance) was diluted to 1:1500. For in situ hybridization, an anti-DIG antibody conjugated to alkaline phosphatase (Roche) was diluted to 1:5000.
6
Protein Extraction and Western Blot Analysis
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C2C12 cells were lysed using a modified RIPA buffer (50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1 mM ethylenediaminetetraacetic acid (EDTA), 1% NP-40, 1% glycerol, and a cocktail of protease inhibitors) for whole-cell extracts, or with a modified NE-PER Nuclear and Cytoplasmic Extraction Kit, as per the manufacturer’s instructions (Thermo Scientific, Waltham, MA, USA). Extracted proteins were then separated via SDS-PAGE and then transferred to a 0.45 μM polyvinylidene fluoride (PVDF) membrane (Millipore, Burlington, MA, USA) on a TRANS-BLOT SD apparatus (Bio-Rad, Hercules, CA, USA). Membranes were blocked with Tris-buffered saline plus 0.1% Tween-20 (TBST) containing 5% skim milk for 1 h at room temperature. Membranes were then incubated overnight at 4 °C in primary antibody made with the blocking solution. The primary antibodies used in this study included mouse SATB2 (ab51502; Abcam, Cambridge, UK; 1:1000), rabbit cleaved caspase 7 (9491S; Cell Signaling, Danvers, MA, USA; 1:1000), mouse myosin heavy chain (Development Studies Hybridoma Bank, DSHB Iowa City, IA, USA; MHC; 1:250), rabbit lamin A/C (2032; Cell Signaling; 1:1000), mouse tubulin (DSHB; 1:200), and mouse glyceraldehyde 3-phosphate dehydrogenase (#2118; GAPDH; 1:4000). Western blots were performed on three independent cell preparations in all cases (n = 3).
7
Immunolabeling Neuronal Markers in Tissue
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Anti-GFP (1:1,000, Aves, GFP-1020), anti-RFP (1:1,000, Rockland Pharmaceuticals, 600-401-379), anti-mCherry (1:500, OriGene AB0081-500), anti-mKATE2 for Brainbow 3.0 (a gift from D. Cai), anti-SATB2 (1:20, Abcam ab51502), anti-CTIP2 (1:100, Abcam 18465), anti-CUX1 (1:100, SantaCruz 13024), anti-LDB2 (1:200, Proteintech 118731-AP), anti-FOG2 (1:500, SantaCruz m-247), anti-LHX2 (1:250, Millipore-Sigma ABE1402) and anti-TLE4 (1:300, Santa Cruz sc-365406) were used.
8
Immunohistochemical Labeling of Neuronal Subtypes
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For ICC, slices were incubated for 20 min in 50% methanol with 1% H2O2 to quench endogenous peroxidases. The slices were rinsed with PBS supplemented with 0.2% Triton X-100 (T-PBS) and blocked for 1 hr in T-PBS, 0.4 g merthiolate and 1% normal goat serum before adding primary antibodies in respective dilutions for 2–3 days at 4°C. Slices were rinsed three times in T-PBS and then incubated in Alexa-fluorophore-conjugated secondary antibodies in T-PBS, 0.4 g merthiolate and 1% normal goat serum overnight at 4°C. Slices were rinsed three times with T-PBS before staining with DAPI (1:5000 in PBS for 2–5 min). After four final rinses (3x PBS, 1x Ampuwa water), slices were mounted in Fluoromount G (SouthernBiotech) on glass slides. Primary and secondary antibodies were used at the following dilutions: mouse anti-Satb2 for excitatory neurons (1:100, Abcam, ab51502, RRID:AB_882455 ), chicken anti-MAP2 for the somatodendritic compartment of neurons (1:500, Abcam, ab5392, RRID:AB_2138153 ); chicken anti-Calretinin (1:500, Merck, AB1550, RRID:AB_90764 ); goat anti-mouse IgG Alexa 568 (1:500, Thermo Fisher, A-11004, RRID:AB_2534072 ); goat anti-chicken Alexa 488 (1:500, Thermo Fisher, A-11039, RRID:AB_142924 ) and donkey anti-goat Alexa 568 (1:500, Thermo Fisher, A-11057, RRID:AB_142581 ).
9
Immunofluorescence Staining of Mouse Tissue Sections
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Mouse tissues were fixed in 4% PFA in PBS overnight at 4 °C and embedded in paraffin. For immunofluorescence, tissue sections of 5 μm were cut, dewaxed and rehydrated. Antigen retrieval was performed by microwaving the sections on 0.01 M sodium citrate buffer (pH 6.0) for 4 min. Tissue sections were blocked in 5% normal donkey serum (NDS) for 30 min after sensing with PBS. Tissue sections then were incubated with primary antibodies diluted in 5% NDS overnight at 4 °C. Antibodies used were: mouse anti-SATB2 (1:10, ab51502, abcam), rat anti-CTIP2 (1:100, ab18465, abcam), and rabbit anti-MBP (1:500; 78896; Cell Signaling technology). After washing with PBS, sections were incubated with Alexa Fluor 488 donkey anti-mouse IgG (1:300; 715-545-150; Jackson ImmunoResearch) or R-Phycoerythrin AffiniPure F(ab’)2 Fragment Donkey Anti-Rat IgG (1:300; 712-116-153; Jackson ImmunoResearch) for 1 h and mounted using Vectorshield mounting media with DAPI (H1200, Vector Laboratories). Images were captured using a Zeiss Axio Imager microscope (Carl Zeiss GmbH, Oberkochen, Germany) and an installed AxioCam HRc camera (Carl Zeiss GmbH) with image acquisition via Zeiss Zen Pro software (v.2.3; Carl Zeiss GmbH).
10
Comprehensive Antibody Panel for Neural Cell Characterization
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Antibodies for CTIP2 (ab18465, 1:500), CUX1 (ab54583, 1:500), PE-conjugated anti EGFR (ab231, 1:50), LGALS1 (ab25138, 1:1,000), Lin28 (ab46020, 1:1,000), PHH3 (ab5176, 1:250), PLZF (ab104854, 1:100), POU3F2 (ab94977, 1:1,000), SATB2 (ab51502, 1:50), SOX1 (1:1,000), SOX2 (ab79351, 1:500), TBR1 (ab31940, 1:200), TBR2 (ab23345, 1:200) were from Abcam. Antibodies for BrdU (347580, 50 μl per test), KI67 (556003, 1:1,000), phycoerythrin-conjugated (PE) SSEA-3 (560237, 20 μl per test), PE-conjugated F11R (552556; 20 μl per test), Alexa Fluor 647-conjugated TRA-1-60-647 (560850, 5 μl per test), Alexa Fluor 647-conjugated TUJ1 (560340, 1:500) were from BD Biosciences. Antibodies for DCX (AB2253, 1:5,000), O4 (MAB345, 1:25), RELN (MAB5364, 1:200), Tyrosine Hydroxylase (TH, AB152, 1:500) were purchased from Millipore. Antibodies for FABP7 (51010-1-AP, 1:100), S100B (15146-1-AP, 1:100) were from ProteinTech. Antibodies for AP2α (3B5 concentrated, 1:100) and PAX6 (supernatant, 1:16) were from DSHB. Antibody for NESTIN (MO15012, 1:500) was from Neuromics. Antibody for GFAP (Z0334, 1:2,000) was from DAKO. Antibody for β-3-Tubulin (PRB-435P, 1:1,000) was from Covance. Antibody for GLAST (ACSA-1; 130-095-822, 1:10) was from Miltenyi Biotec. Antibody for OTX2 (AF1979; 1:40) was from R&D. Antibody for FOXA2 (SC-6554; 1:100) was from Santa Cruz Biotechnology.
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