Anti-gamma Tubulin antibody - Centrosome Marker (ab16504)

Overview

  • Product nameAnti-gamma Tubulin antibody - Centrosome Marker
    See all gamma Tubulin primary antibodies
  • Description
    Rabbit polyclonal to gamma Tubulin - Centrosome Marker
  • SpecificityDoes not react with alpha or beta tubulin. Immunogen sequence found in both gamma tubulin 1 and gamma tubulin 2.
  • Tested applicationsSuitable for: ICC/IF, IP, IHC - Wholemount, IHC-Fr, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human, Zebrafish
    Predicted to work with: Rat, Dog, Xenopus laevis
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human gamma Tubulin.

    (Peptide available as ab17097.)

  • Positive control
    • WB: HeLa Whole Cell Lysate, HeLa Nuclear Cell Lysate, A431 Whole Cell Lysate ICC: SK-N-SH cells

Properties

Applications

Our Abpromise guarantee covers the use of ab16504 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF Use a concentration of 5 µg/ml.
IP Use at an assay dependent concentration.
IHC - Wholemount Use at an assay dependent concentration.
IHC-Fr Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 48 kDa (predicted molecular weight: 48 kDa).

Abcam recommends using milk as the blocking agent.

Target

Anti-gamma Tubulin antibody - Centrosome Marker images

  • ICC/IF image of ab16504 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab16504, 5µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

  • SK-N-SH cells were fixed in 4% paraformaldehyde, permeabilized in 0.5% Triton X-100 and incubated for 1 hour with  ab16504 (1/300). ab16504 staining is localized to the centrosome (red). The cells were counterstained with DAPI (blue). 100x magnification. The cells were blocked with 5% fetal bovine serum.

  • SK-N-SH cells were fixed in 4% paraformaldehyde, permeabilized in 0.5% Triton X-100 and incubated for 1 hour with ab16504. The antibody clearly labels the centrosome (red). The cells were counterstained with DAPI (blue). The cells were blocked in 5% BSA.

  • ICC/IF image of ab16504 stained Hek293 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab16504, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    The cause of the background staining is uncertain, although a cytokeratin exisits of a similar molecular weight and amino acid sequence to that of the immunogen used to raise the antibody.
  • All lanes : Anti-gamma Tubulin antibody - Centrosome Marker (ab16504) at 1 µg/ml

    Lane 1 : Marker
    Lane 2 : Zebrafish brain homogenate at 20 µg
    Lane 3 : Zebrafish liver homogenate at 20 µg
    Lane 4 : HeLa (Human epithelial carcinoma cell line) whole cell lysate at 20 µg

    Secondary
    Goat polyclonal to Rabbit IgG – H&L – Pre-Adsorbed (HRP) at 1/6000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 48 kDa
    Observed band size : 48 kDa


    Exposure time : 5 minutes
  • ab16504 staining gamma Tubulin in E8 murine embryos by IHC - Wholemount.
    Samples were incubated with ab16504 at a 1/150 dilution for 12 hours at 4°C in 0.1% Triton/PBS. ab6717 Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (FITC) was used as the secondary at a 1/250 dilution.

    See Abreview

References for Anti-gamma Tubulin antibody - Centrosome Marker (ab16504)

This product has been referenced in:
  • Bancroft J  et al. Chromosome congression is promoted by CENP-Q- and CENP-E-dependent pathways. J Cell Sci 128:171-84 (2015). ICC/IF ; Human . Read more (PubMed: 25395579) »
  • Sikkema WK  et al. Regulation of mitotic cytoskeleton dynamics and cytokinesis by integrin-linked kinase in retinoblastoma cells. PLoS One 9:e98838 (2014). ICC/IF ; Human . Read more (PubMed: 24911651) »

See all 9 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application IHC - Wholemount
Sample Mouse Embryo (mouse embryo E8)
Specification mouse embryo E8
Username

Ms. Patricia Ybot

Verified customer

Submitted Feb 23 2012

Este anticuerpo debe funcionar perfectamente en muestras frescas, siempre que estas se conserven frescas, y la tinción se lleve a cabo enseguida para evitar una posible degradación de las proteínas. Con el mismo propósito debe llevarse a cabo la tinció...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Methanol
Permeabilization No
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Dec 11 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Sample Human Cell lysate - whole cell (U2OS cells)
Total protein in input 600 µg
Specification U2OS cells
Immuno-precipitation step Protein A/G
Username

Abcam user community

Verified customer

Submitted Jun 21 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (U2OS cells)
Loading amount 30 µg
Specification U2OS cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Abcam user community

Verified customer

Submitted May 10 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (embryo)
Specification embryo
Fixative Paraformaldehyde
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 6%
Username

Abcam user community

Verified customer

Submitted Apr 30 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (U2OS)
Specification U2OS
Fixative Methanol
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Abcam user community

Verified customer

Submitted Jan 18 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (primary human monocyte-derived macrophages and Hel)
Loading amount 10 µg
Specification primary human monocyte-derived macrophages and Hel
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Abcam user community

Verified customer

Submitted Aug 04 2006

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Hela)
Specification Hela
Fixative Paraformaldehyde
Blocking step BSA as blocking agent for 45 minute(s) · Concentration: .5%
Username

Abcam user community

Verified customer

Submitted Jul 27 2006

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"