The following unconjugated primary antibodies were used: mouse monoclonal (mAb) anti-GluN2B and anti-PSD-95 (1:2000, #75–097 clone 59/20 and #75028, Neuromab, Davis, CA, USA); rabbit polyclonal (rAb) anti-GluN2A (1:1000) and mAb anti-alpha-tubulin (1:10,000, #M264 and #T9026 respectively, Sigma-Aldrich, St. Louis, MO, USA); rAb anti-MAP2 (1:500, #AB5622, Merck-Millipore, Billerica, MA, USA); mAb anti-HIV1 TAT antibody (1:200, #63957, Abcam, Cambridge, UK). Goat AlexaFluor 488- and 555-conjugated anti-mouse and anti-rabbit secondary antibodies (both 1:250, #A11029 and #A21429, Life Technologies, Monza, Italy) were used for immunofluorescence studies, while goat anti-mouse or anti-rabbit horseradish peroxidase (HRP)-conjugated antibodies were bought from Bio-Rad (both 1:10,000, #172–1011 and #172–1019, Hercules, CA, USA) for western blot (WB) analysis.
Anti alpha tubulin mab
Manufactured by Merck Group
Sourced in United States
Anti-alpha-tubulin mAb is a monoclonal antibody that recognizes the alpha-tubulin subunit of microtubules. It is a commonly used tool in cell biology research for the detection and visualization of microtubule structures within cells.
Automatically generated - may contain errors
Lab products found in correlation
Anti glun2a, by Merck Group (1 mentions)
Horseradish peroxidase hrp conjugated antibody, by Bio-Rad (1 mentions)
Anti flag mab, by Merck Group (1 mentions)
Bradford assay dye reagent, by Bio-Rad (1 mentions)
Ecl western detection system, by Cytiva (1 mentions)
Pvdf membrane, by Bio-Rad (1 mentions)
Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 cell culture medium, by Merck Group (1 mentions)
Ova323 339 peptide, by GenScript (1 mentions)
Ripa lysis buffer, by Santa Cruz Biotechnology (1 mentions)
5 protocols using anti alpha tubulin mab
1
Antibody Characterization for Neuroscience
2
Antibody Characterization for Signaling Pathways
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The following antibodies were used: monoclonal antibody (mAb) anti-α-calcium/calmodulin-dependent kinase II (αCaMKII), polyclonal antibody (pAb) anti-GluN2A, pAb anti-CREB, pAb anti-p-CREB (Ser-133), and mAb anti-Myc were purchased from Millipore (Billenca, MA, USA); mAb anti-Meox2 was purchased from Abcam (Cambridge, MA, USA); pAb anti-p44/42 MAPK, pAb anti-p-p44/42 MAPK (Thr202/Tyr204) were purchased from Cell Signaling (Danvers, MA, USA); mAb anti-GFP, mAb anti-GST and anti-PSD-95 were purchased from NeuroMab (Davis, CA, USA); mAb anti-alpha-Tubulin and mAb anti-Flag were purchased from Sigma-Aldrich (St. Louis, MO, USA); mAb anti-p21 were purchased from BD Biosciences (NJ); pAb antihistone H3 and pAb anti-RNF10 were purchased from Proteintech (Chicago, MI, USA, USA); mAb anti-JL8 was purchased from Clontech (Mountain View, CA, USA); pAb anti-HA was purchased from Santa Cruz Biotechnology (Dallas, TX, USA) p-RNF10(S31) was custom generated from Primm (Cambridge, MA, USA). Peroxidaseconjugated secondary anti-mouse Ab and peroxidaseconjugated secondary anti-rabbit Ab was purchased from Bio-Rad (Hercules, CA, USA). AlexaFluor secondary Abs were purchased from Invitrogen (Carlsbad, CA, USA).
3
Western Blot Analysis of Autophagy Markers
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Tissue samples were lysed in lysis buffer containing 1 % Triton X-100, 10 mM Tris–HCl (pH 7.5), 150 mM NaCl, 5 mM EDTA, 1 mM Na3VO4 and 75 U of aprotinin and allowed to stand for 20 min at 4 °C. The tissue suspension was mechanically disrupted by Dounce homogenization (ten strokes). The lysate was centrifuged for 5 min at 1300×g to remove nuclei and large cellular debris. After evaluation of the protein concentration by Bradford dye reagent assay (Bio-Rad, 500-0006), the lysate was subjected to 8 % (for AMBRA1) or 15 % (for other antigens) sodium-dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The proteins were electrophoretically transferred onto polyvinylidene difluoride (PVDF) membranes (Bio-Rad, 162-0177). Membranes were blocked with 5 % defatted dried milk in TBS, containing 0.05 % Tween-20 and probed with rabbit polyclonal anti-LC3 antibody (MBL Int Corporation, PD014), with rabbit polyclonal anti-AMBRA1 (NOVUS, NBP1-07124), with rabbit polyclonal anti-p62 SQSTM1/sequestosome antibody (Sigma, P0067) or with anti-alpha-tubulin Mab (Sigma, T6199) Bound antibodies were visualized with horseradish peroxidase (HRP)-conjugated anti-rabbit IgG (Sigma, A1949) or anti-mouse IgG (Sigma, A9044) and immunoreactivity assessed by chemiluminescence reaction, using the ECL Western detection system (Amersham, RPN2106).
4
T Cell Activation and Signaling Protocol
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Antibodies against mouse and rat cell surface molecules conjugated to fluorochromes or biotin, and fluorescent streptavidin conjugates were purchased from eBioscience, BD Biosciences or Biolegend (S1 Table ). Antibodies against phospho STAT5 (Y694), phospho ZAP70, phospho LAT, STAT5, ZAP70, LAT and FOXO1 were from Cell Signaling Technology or Santa Cruz Biotechnology Inc. Anti-mouse CD3 mAb (2C11) and goat anti-hamster antibodies were from BD Pharmingen Biosciences. Carboxyfluorescein succinimidyl ester (CFSE) was from Invitrogen. RPMI-1640 cell culture medium, fetal bovine serum (FBS), 4G10 that recognizes phospho-tyrosine on tyrosine phosphorylated proteins and anti-alpha-tubulin mAb were from Sigma Aldrich. Nocodazole, colchicine, cytochalasin D and latrunculin B were obtained from Calbiochem. OVA323-339 peptide (ISQAVHAAHAEINEAGR) was custom synthesized by Genscript (New Jersey, USA).
5
Western Blot Analysis of Cell Lysates
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