Antibodies used were as follows: anti-Emerin (5430, Cell Signaling Technology, Danvers, MA, USA 1:500 for IF, 1:1000 for IB), anti-Lemd2 (PA553589, Thermo Fisher Scientific, Waltham MA, USA 1:300 for IF, 1:1000 for IB), anti-Ankle2 (GTX120698, Genetex, Irvine, CA, United States 1:200 for IF, 1:1000 for IB), and anti-TMPO (L3414-.2ML, Sigma-Aldrich, Saint Louis, MO 1:500 for IF, 1:1000 for IB), anti-GAPDH (glyceraldehyde-3-phosphate dehydrogenase) (D16H11, Cell Signaling Technology, 1:4000 for IB), and anti–Gamma-Tubulin (T6557, Sigma-Aldrich, 1:3000 for IB). Fluorescent secondary antibodies used were Alexa Fluor 488 (Cat# A32766, Molecular Probes, Thermo Fisher Scientific 1:200 for IF) and 594 (Cat# A32754, Molecular Probes, Thermo Fisher Scientific, 1:200 for IF), IRDye® 800CW Donkey anti-Mouse IgG Secondary Antibody (926-32212, LiCor Bioscience, Lincoln, NE, USA), and IRDye® 680RD Donkey anti-Rabbit IgG Secondary Antibody (926-68073, LiCor Bioscience).
Anti gamma tubulin
Manufactured by Merck Group
Sourced in United States
Anti-gamma tubulin is a lab equipment product that is used to detect and analyze the gamma-tubulin protein. Gamma-tubulin is a critical component of the microtubule-organizing center (MTOC), which is responsible for the organization and assembly of microtubules within cells. The anti-gamma tubulin product can be used in various cell and molecular biology techniques, such as immunofluorescence and Western blotting, to study the distribution and function of gamma-tubulin in different cell types and conditions.
Automatically generated - may contain errors
Lab products found in correlation
Anti gapdh, by Cell Signaling Technology (1 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (1 mentions)
Anti emerin, by Cell Signaling Technology (1 mentions)
Irdye 800cw donkey anti mouse igg secondary antibody, by LI COR (1 mentions)
Irdye 680rd donkey anti rabbit igg secondary antibody, by LI COR (1 mentions)
Anti insulin, by Agilent Technologies (1 mentions)
Af3628, by Abcam (1 mentions)
Anti e cadherin, by Cell Signaling Technology (1 mentions)
Anti zeb1, by Santa Cruz Biotechnology (1 mentions)
Anti e cadherin, by Abcam (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
Anti cd31, by R&D Systems (1 mentions)
Anti lyve 1, by Abcam (1 mentions)
Anti sv40 t ag, by Santa Cruz Biotechnology (1 mentions)
Alexa fluor fluorophores, by Thermo Fisher Scientific (1 mentions)
Anti α tubulin, by Abcam (1 mentions)
Hoechst 33342 dye, by Thermo Fisher Scientific (1 mentions)
Anti eea1, by Santa Cruz Biotechnology (1 mentions)
Anti rab11a, by Santa Cruz Biotechnology (1 mentions)
Anti acetylated tubulin clone 6 11b 1, by Merck Group (1 mentions)
6 protocols using anti gamma tubulin
1
Antibody Immunofluorescence and Western Blot Protocol
2
Comprehensive Antibody Profiling for Immunoblotting and Immunohistochemistry
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The following antibodies were used in immunoblotting: anti-E-cadherin (Cell Signaling, 3195, 1:1000), anti-Zeb1 (Santa Cruz, sc-25388, 1:1000), anti-Beta-Actin (Cell Signaling, 4970S, 1:1000), and anti-Gamma-Tubulin (Sigma, T6557, 1:5000). The following antibodies were used for immunohistochemistry: anti-SV40 T Ag (Santa Cruz, sc-20800, 1:50), anti-Insulin (DAKO, A056401, 1:1000), anti-CD31 (R&D Systems, AF3628, 1:15), anti-Lyve1 (Abcam, ab14917, 1:100), anti-E-cadherin (Abcam, ab76055, 1:200), anti-Glucagon (Millipore, AB932, 1:200), anti-SMA (Abcam, ab5694, 1:100), anti-CK19 (Abcam, ab52625, 1:400).
3
Immunofluorescence Labeling of Organelles
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Immunofluorescence experiments utilized 4% bovine serum albumin (BSA, Sigma–Aldrich, A7030) in PBS as blocking reagent and antibody diluent. Where necessary, fixed cells were permeabilized with 0.1% Triton-X 100 in PBS prior to blocking. Dextran labeled cells were not permeabilized, but utilized 0.05M glycine in the blocking media. Actin filaments were labeled with Alexa-647 conjugated Phalloidin dyes (ThermoFisher Scientific, A22287) per the manufacturer protocol. Primary antibodies used were anti-Alpha tubulin (1:500, Abcam ab7291), anti-ARL13B (1:250, ProteinTech 17711-1-AP), anti-gamma tubulin (1:250, Sigma–Aldrich T5326), anti-EEA1 (1:200, SantaCruz Biotechnology sc-6414), anti-LAMP3 (1:200, ThermoFisher Scientific, MA1-35272), anti-Rab9 (1:500, ThermoFisher Scientific, MA3-067), anti-Rab4 (1:250, ThermoFisher Scientific, PA3-912), anti-Rab11a (1:250, SantaCruz Biotechnology sc-166912), and anti-Rab7 (1:100, ThermoFisher Scientific, PA5-52369). Secondary antibodies used were donkey anti-rabbit, anti-mouse or anti-goat and conjugated to AlexaFluor fluorophores (ThermoFisher Scientific, 1:1000 dilution). Primary and secondary antibodies were diluted in 4% BSA in PBS. DNA counterstaining utilized Hoechst33342 dye (ThermoFisher Scientific, H1399), diluted 1:5000 in PBS.
4
Antibody panel for cellular analysis
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Anti-GFP (Roche, 11 814 460 001), anti-HA (mouse hybridoma 12CAS), anti-Acetylated tubulin (Clone 6—11B-1, T6793, Sigma), anti-gamma tubulin (Clone GTU-88, Sigma), anti-A cyclase III (C-20, sc-588, Santa Cruz Biotechnology INC., Dallas, TX, USA), anti-Rab3IP (S-14, sn-162069, Santa Cruz Biotechnology Inc., Dallas, TX, USA).
5
Immunoprecipitation and Immunohistochemistry of Transfected HEK293 Cells
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Authenticated HEK293 cells were purchased from ATCC and tested for mycoplasma by PCR. HEK293 cells were grown in six well plates and transfected with 4 µg of each DNA with lipofectamine 2000 (Invitrogen) for 6 hr according to manufacturers instructions.
For immunohistochemistry, cells were fixed 48 hr after transfection in 4% paraformaldehyde in PBS for 20 min at room temperature, washed with PBS, and permeabilized in 0.2% Triton X-100 in PBS for 5 min at room temperature. The protocol was followed as in Supp. Materials and methods.
For immunoprecipitation, cells were grown for 24 hr after transfection and then proteins were extracted following standard methods (Supp. Materials and methods). The eluate from antibody beads (30 µl) was loaded in 10% polyacrylamide gels and proteins were detected by Western blots (standard conditions) using anti-myc (1/20,000, SC-40, SCBT), anti-HA (1/10,000, 3F10, Roche) and anti-β-catenin (1/8000, Sigma, C7207), to detect the co-immunoprecipitated proteins. Antibodies used on Western blots inFigure 3—figure supplement 1B are anti human Tcf7l2 (N-20, SCBT) and anti-gamma tubulin (T9026, Sigma) HRP coupled secondary antibodies (1/2,000, sigma) were used and blots were developed using an ECL kit (Promega).
For immunohistochemistry, cells were fixed 48 hr after transfection in 4% paraformaldehyde in PBS for 20 min at room temperature, washed with PBS, and permeabilized in 0.2% Triton X-100 in PBS for 5 min at room temperature. The protocol was followed as in Supp. Materials and methods.
For immunoprecipitation, cells were grown for 24 hr after transfection and then proteins were extracted following standard methods (Supp. Materials and methods). The eluate from antibody beads (30 µl) was loaded in 10% polyacrylamide gels and proteins were detected by Western blots (standard conditions) using anti-myc (1/20,000, SC-40, SCBT), anti-HA (1/10,000, 3F10, Roche) and anti-β-catenin (1/8000, Sigma, C7207), to detect the co-immunoprecipitated proteins. Antibodies used on Western blots in
6
Histological and Immunostaining Analysis of Bone Development
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H&E, Safranin-O, Picrosirius red staining and von Kossa staining were carried out using standard methods 33 . Staining of long bones with Alizarin red and Alcian blue was carried out as previously described 33 . Immunofluorescent staining and TUNEL assays were performed as previously described 34 . Anti-acetylated tubulin (Sigma; T6793, 1:1,000), anti-gamma tubulin (Sigma; T5326, 1:1,000), anti-phospho Histone H3 (Millipore; #06-570, 1:100), anti-SOX9 (Santa cruz biotechnology; sc-20095, 1:100), anti-phospho ERK1/2 (Cell signaling technology; #4376, 1:100) antibodies were used for immunostaining. Slides were imaged under an Olympus FluoView FV1000 laser scanning confocal microscope using the software FV10-ASW Viewer (version 4.2).
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