Overview

  • Product name
    Anti-gamma Tubulin antibody - Centrosome Marker
    See all gamma Tubulin primary antibodies
  • Description
    Rabbit polyclonal to gamma Tubulin - Centrosome Marker
  • Host species
    Rabbit
  • Specificity
    Does not react with alpha or beta tubulin. Immunogen sequence found in both gamma tubulin 1 and gamma tubulin 2.
  • Tested applications
    Suitable 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.

    Read Abcam's proprietary immunogen policy (Peptide available as ab17097.)

  • Positive control
    • WB: Recombinant Human gamma Tubulin protein (denatured) (ab115710), 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

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).

  • Lane 1 - 6 : gamma Tubulin antibody - Centrosome Marker (ab11317) at 1 ug/ml

    Lane 1 : HeLa Whole Cell Lysate
    Lane 2 : HeLa Nuclear Cell Lysate
    Lane 3 : A431 Whole Cell Lysate
    Lane 4 : HeLa Whole Cell Lysate with gamma Tubulin peptide (38-53) (ab17097)
    Lane 5 : HeLa Nuclear Cell Lysate with gamma Tubulin peptide (38-53) (ab17097)
    Lane 6 : A431 Whole Cell Lysate with gamma Tubulin peptide (38-53) (ab17097)

    Secondary

    Alexa Fluor Goat polyclonal to Rabbit IgG at 1/10000 dilution

    Lysates at 20 ug.
    Blocking peptide at 1 ug/ml.
    Performed under reducing conditions.

    Observed band size : 48kD

  • 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.

  • Paraformaldehyde-fixed mouse embryo tissue stained for gamma Tubulin (red) using ab16504 in immunohistochemical analysis.

    See Abreview

  • 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
    All lanes : 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

This product has been referenced in:
  • Ishikawa KI  et al. p150glued deficiency impairs effective fusion between autophagosomes and lysosomes due to their redistribution to the cell periphery. Neurosci Lett 690:181-187 (2019). Read more (PubMed: 30366015) »
  • Revinski DR  et al. CDC20B is required for deuterosome-mediated centriole production in multiciliated cells. Nat Commun 9:4668 (2018). Read more (PubMed: 30405130) »
See all 21 Publications for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

Filter by Ratings

1-9 of 9 Abreviews

Application
Immunocytochemistry/ Immunofluorescence
Sample
Plasmodium falciparum Cell (Blood stage)
Permeabilization
Yes - 0.1% triton-X 100
Specification
Blood stage
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: RT°C
Fixative
Paraformaldehyde

Julien Guizetti

Verified customer

Submitted May 11 2018

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

Patricia Ybot

Verified customer

Submitted Feb 23 2012

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

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

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%

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%

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%

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%

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%

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"

Sign up