Anti-gamma Tubulin antibody [EPR16793] - Centrosome Marker

11 product images

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  Immunocytochemistry/ Immunofluorescence - Anti-gamma Tubulin antibody [EPR16793] - Centrosome Marker (ab179503)

  Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling gamma Tubulin with ab179503 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/400 dilution (green). Cytoplasmic and centrosome staining on HeLa cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
  The negative controls are as follows;
  1. ab179503 at 1/500 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
  2. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

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  Western blot - Anti-gamma Tubulin antibody [EPR16793] - Centrosome Marker (ab179503)

  Blocking/Dilution buffer: 5% NFDM/TBST.

  All lanes: Western blot - Anti-gamma Tubulin antibody [EPR16793] - Centrosome Marker (ab179503) at 1/20000 dilution

  All lanes: Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysates at 20 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

  Predicted band size: 51 kDa

  Observed band size: 48 kDa

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  Western blot - Anti-gamma Tubulin antibody [EPR16793] - Centrosome Marker (ab179503)

  Blocking/Dilution buffer: 5% NFDM/TBST.

  All lanes: Western blot - Anti-gamma Tubulin antibody [EPR16793] - Centrosome Marker (ab179503) at 1/2000 dilution

  Lane 1: Human fetal brain lysates at 10 µg

  Lane 2: Human fetal heart lysates at 10 µg

  Lane 3: Human fetal kidney lysates at 10 µg

  Secondary

  All lanes: Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

  Predicted band size: 51 kDa

  Observed band size: 48 kDa

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  Western blot - Anti-gamma Tubulin antibody [EPR16793] - Centrosome Marker (ab179503)

  Blocking/Dilution buffer: 5% NFDM/TBST.

  All lanes: Western blot - Anti-gamma Tubulin antibody [EPR16793] - Centrosome Marker (ab179503) at 1/2000 dilution

  Lane 1: HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates at 20 µg

  Lane 2: MDCK (Canine kidney cell line) whole cell lysates at 20 µg

  Lane 3: MDBK (Bovine kidney cell line) whole cell lysates at 20 µg

  Lane 4: COS-1 (African green monkey kidney fibroblast-like cell line) whole cell lysates at 20 µg

  Lane 5: Zebrafish lysates at 20 µg

  Lane 6: Xenopus (X. tropicalis) muscle lysates at 20 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

  Predicted band size: 51 kDa

  Observed band size: 48 kDa

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  Western blot - Anti-gamma Tubulin antibody [EPR16793] - Centrosome Marker (ab179503)

  Blocking/Dilution buffer: 5% NFDM/TBST.

  All lanes: Western blot - Anti-gamma Tubulin antibody [EPR16793] - Centrosome Marker (ab179503) at 1/20000 dilution

  Lane 1: UMNSAH/DF-1 (Transformed chicken embyronic fibroblast cells) whole cell lysates at 20 µg

  Lane 2: BHK (Hamster kidney fibroblast cells) whole cell lysates at 20 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

  Predicted band size: 51 kDa

  Observed band size: 48 kDa

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  Western blot - Anti-gamma Tubulin antibody [EPR16793] - Centrosome Marker (ab179503)

  Blocking/Dilution buffer: 5% NFDM/TBST.

  All lanes: Western blot - Anti-gamma Tubulin antibody [EPR16793] - Centrosome Marker (ab179503) at 1/2000 dilution

  Lane 1: Mouse brain lysates at 10 µg

  Lane 2: Mouse spleen lysates at 10 µg

  Lane 3: Rat brain lysates at 10 µg

  Lane 4: Rat heart lysates at 10 µg

  Lane 5: Rat kidney lysates at 10 µg

  Lane 6: C6 (Rat glial tumor cells) whole cell lysates at 10 µg

  Lane 7: RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysates at 10 µg

  Lane 8: PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates at 10 µg

  Lane 9: NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysates at 10 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

  Predicted band size: 51 kDa

  Observed band size: 48 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-gamma Tubulin antibody [EPR16793] - Centrosome Marker (ab179503)

  Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling gamma Tubulin with ab179503 at 1/250 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasmic staining on epithelial cells of mouse testis is observed. Counter stained with Hematoxylin.
  

  
  Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

  Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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  Immunoprecipitation - Anti-gamma Tubulin antibody [EPR16793] - Centrosome Marker (ab179503)

  Gamma Tubulin was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell extract with ab179503 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab179503 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG was used as secondary antibody at 1/1500 dilution. Lane 1: HeLa whole cell extract. Lane 2: PBS instead of HeLa whole cell extract.

  Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  All lanes: Immunoprecipitation - Anti-gamma Tubulin antibody [EPR16793] - Centrosome Marker (ab179503)

  Predicted band size: 51 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-gamma Tubulin antibody [EPR16793] - Centrosome Marker (ab179503)

  Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling gamma Tubulin with ab179503 at 1/250 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasmic staining on tumor cells of breast carcinoma is observed. Counter stained with Hematoxylin.
  

  
  Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

  Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-gamma Tubulin antibody [EPR16793] - Centrosome Marker (ab179503)

  Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling gamma Tubulin with ab179503 at 1/250 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasm staining on epithelial cells of rat testis is observed. Counter stained with Hematoxylin.
  

  Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

  Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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  Flow Cytometry (Intracellular) - Anti-gamma Tubulin antibody [EPR16793] - Centrosome Marker (ab179503)

  Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed UMNSAH/DF-1 (Transformed chicken embyronic fibroblast cells) cells labeling gamma Tubulin with ab179503 at 1/80 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.