| gamma Tubulin Antibody (MA1-850) in IFImmunofluorescent analysis of g-Tubulin using anti-g-Tubulin monoclonal antibody (Product # MA1-850, Clone 4D11) (shown in green) in HeLa (A) and U2-OS cells (B). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were then blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with a mouse monoclonal antibody recognizing g-Tubulin (Product # MA1-850), at a dilution of 1:50 for at least 1 hour at room temperature. Cells were washed with PBS and incubated with DyLight 488 goat-anti-mouse secondary antibody (Product # 35503) at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DY-547 phalloidin, nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan at 20X magnification.gamma Tubulin Antibody (MA1-850) in WBWestern blot analysis was performed by loading 50 µg of various whole cell lysates onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% Milk/TBST for at least 1 hour. Membranes were probed with a mouse monoclonal antibody recognizing g-Tubulin (Product # MA1-850, Clone 4D11) at a dilution of 1:1000 overnight at 4°C on a rocking platform. Membranes were washed in TBS-0.1%Tween 20 and probed with a goat anti-mouse-HRP secondary antibody (Product # 31430) at a dilution of 1:20,000 for at least one hour. Membranes were washed and chemiluminescent detection performed using Pierce Super Signal West Dura (Product # 34075).gamma Tubulin Antibody (MA1-850) in IHCImmunohistochemistry was performed on cancer biopsies of deparaffinized human Breast carcinoma tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a mouse monoclonal antibody recognizing gamma Tubulin (Product # MA1-850) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.gamma Tubulin Antibody (MA1-850) in IHCImmunohistochemistry was performed on normal deparaffinized human Colon tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing gamma Tubulin (Product # MA1-850) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.gamma Tubulin Antibody (MA1-850) in WBKnockdown of gamma Tubulin was achieved by transfecting HeLa cells with gamma Tubulin specific validated siRNA (Silencer® select Cat # s14502). Western blot analysis (Fig a) was performed using modified whole cell extract (1% SDS) from the gamma Tubulin knock down cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Anti-gamma Tubulin mouse monoclonal antibody (Product # MA1-850, 1:1000) and Goat anti-Mouse IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Product # A28177, 1:5000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig b). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to gamma Tubulin. |
| 1.Journal of endocrinology and diabetesTopiramate Protects Pericytes from Glucotoxicity: Role for Mitochondrial CA VA in Cerebromicrovascular Disease in Diabetes."MA1-850 was used in western blot to study the role for mitochondrial CA VA in cerebromicrovascular disease in diabetes and protection of pericytes from glucotoxicity by topiramate"AuthorsPatrick P,Price TO,Diogo AL,Sheibani N,Banks WA,Shah GN2.CirculationNucleoside Diphosphate Kinase-C Suppresses cAMP Formation in Human Heart Failure."MA1850 was used in western blot to demonstrate that nucleoside diphosphate kinase-C suppresses cAMP formation in human heart failure"AuthorsAbu-Taha IH,Heijman J,Hippe HJ,Wolf NM,El-Armouche A,Nikolaev VO,Schäfer M,Würtz CM,Neef S,Voigt N,Baczkó I,Varró A,Müller M,Meder B,Katus HA,Spiger K,Vettel C,Lehmann LH,Backs J,Skolnik EY,Lutz S,Dobrev D,Wieland T3.The Journal of biological chemistryNLRP7, a nucleotide oligomerization domain-like receptor protein, is required for normal cytokine secretion and co-localizes with Golgi and the microtubule-organizing center."MA1-850 was used in Immunofluorescence to study the effect of NLRP7 mutations on peripheral blood mononuclear cell's ability to secrete IL-1β and TNF in response to LPS."AuthorsMessaed C,Akoury E,Djuric U,Zeng J,Saleh M,Gilbert L,Seoud M,Qureshi S,Slim R4.Human molecular geneticsSynphilin-1 attenuates neuronal degeneration in the A53T alpha-synuclein transgenic mouse model."MA1-850 was used in immunohistochemistry to investigate the neuroprotective effect of synphilin-1 in the A53T alpha-synuclein transgenic mouse model"AuthorsSmith WW,Liu Z,Liang Y,Masuda N,Swing DA,Jenkins NA,Copeland NG,Troncoso JC,Pletnikov M,Dawson TM,Martin LJ,Moran TH,Lee MK,Borchelt DR,Ross CA5.Oncotargetc-MYC drives histone demethylase PHF8 during neuroendocrine differentiation and in castration-resistant prostate cancer."MA1850 was used in western blot to elucidate the regulation of PHF8 and KDM3A during neuroendocrine differentiation and castration-resistant prostate cancer"AuthorsMaina PK,Shao P,Liu Q,Fazli L,Tyler S,Nasir M,Dong X,Qi HH |
QQ:1749072012