Anti-TRA-1-60 (R) [TRA-1-60] antibody (ab16288)

Overview

  • Product nameAnti-TRA-1-60 (R) [TRA-1-60] antibodySee all TRA-1-60 (R) primary antibodies ...
  • Description
    Mouse monoclonal [TRA-1-60] to TRA-1-60 (R)
  • Tested applicationsICC/IF, Flow Cyt, WB, IP, ICC, RIA, IHC-Fr more details
  • Species reactivity
    Reacts with: Rabbit, Human
  • Immunogen

    Human embryonal carcinoma cell line 2102Ep cl.2A6.

  • General notesThe myeloma was fused with a spleen from a Balb/c mouse.

Properties

Applications

Our Abpromise guarantee covers the use of ab16288 in the following tested applications.

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

Application Notes
ICC/IF Use at an assay dependent dilution. PubMed: 20674000
Flow Cyt Use a concentration of 10 - 20 µg/ml.
WB Use a concentration of 1 - 10 µg/ml. Detects a band of approximately 250 kDa (predicted molecular weight: 235 kDa).
IP Use at an assay dependent dilution.
ICC Use at an assay dependent dilution.
RIA Use at an assay dependent dilution.
IHC-Fr Use at an assay dependent dilution.

Target

  • RelevanceThe monoclonal antibody TRA-1-60 is a marker of Human Embryonic Stem, Germ and Carcinoma Cells but Not Mouse Embryonic Stem, Germ or Carcinoma Cells. The core protein recognised by TRA-1-60 has not been identified but the antibody is thought to recognise carbohydrate epitopes associated with a high molecular weight proteoglycan. The TRA antibodies are named after the Battle of Trafalgar; TRA does not stand for "Tumor Rejection Antigen" as is believed by some.
  • Cellular localizationCell Membrane

Anti-TRA-1-60 (R) [TRA-1-60] antibody images

  • ab16288 staining TRA-1-60 (R) in Human H9 cells by Flow Cytometry. Cells were fixed with paraformaldehyde and permeabilized with 0.2% Triton X-100. Cells were incubated with the primary antibody (1/1000) for 1 hour at 4°C. Gating Strategy: Unstained H9 cells.

    See Abreview

  • All lanes : Anti-TRA-1-60 (R) [TRA-1-60] antibody (ab16288) at 10 µg/ml

    Lane 1 : Human Embryonic Stem Cell Lysate
    Lane 2 : Mouse Embryonic Stem Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat F(ab')2 Anti-Mouse IgM mu chain (HRP) (ab5930) at 1/5000 dilution

    Predicted band size : 235 kDa
    Observed band size : 250 kDa (why is the actual band size different from the predicted?)
    As expected, ab16288 recognizes a band in Human Embryonic Stem Cell Lysate but not Mouse Embryonic Stem Cell Lysate. The smear above the main band is due to protein glycosylation. The Western Blot profile is as expected for the TRA-1-60 monoclonal antibody.
  • Anti-TRA-1-60 antibody, ab16288, can be used as a marker of human embryonic stem cells. DAPI staining is shown in the top panel while TRA-1-81 staining with ab16288 is shown in the lower panel.

    See Abreview

  • ab16288 staining  the Embryonic Stem Cell Marker in HUES7 cells by ICC/IF (Immunocytochemistry/immunoflurescence). Cells were fixed with paraformaldehyde, permeabilized with Triton and blocked in 10% serum  for 1 hour at 24°C.  Samples were incubated with primary antibody, dilution 1/500 (1% Serum, 0.1% Triton in PBS), for 1 hour at 37°C. An Alexa Fluor® 647-conjugated Goat polyclonal to mouse IgG, dilution 1/100, was used as secondary antibody.

    See Abreview

References for Anti-TRA-1-60 (R) [TRA-1-60] antibody (ab16288)

This product has been referenced in:
  • Soteriou D  et al. Comparative proteomic analysis of supportive and unsupportive extracellular matrix substrates for human embryonic stem cell maintenance. J Biol Chem 288:18716-31 (2013). Read more (PubMed: 23658023) »
  • Gafni O  et al. Derivation of novel human ground state naive pluripotent stem cells. Nature N/A:N/A (2013). Human . Read more (PubMed: 24172903) »

See all 20 Publications for this product

Product Wall

Application Flow Cytometry
Fixation Paraformaldehyde
Sample Human Cell (H9 human)
Specification H9 human
Gating Strategy Unstained H9 cells
Permeabilization Yes - 0.2% triton x-100
Username

Abcam user community

Verified customer

Submitted Aug 12 2013

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 10 minute(s) · Concentration: 5% · Temperature: 25°C
Sample Human Cell (H9 human)
Specification H9 human
Permeabilization Yes - 0.03% TritonX-100
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jul 11 2013

Application Western blot
Loading amount 40 µg
Gel Running Conditions Reduced Denaturing (12)
Sample Human Cell lysate - whole cell (H9 embryonic stem cells)
Specification H9 embryonic stem cells
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jul 10 2013

Thank you for contacting us. I have checked the WB image information and indeed the figure and the legend information are contradicting. Based on the information that we have in the system, it seems that the WB Lane information was mixed-up and I s...

Read More
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Embryonic Stem Cell, HUES7)
Specification Embryonic Stem Cell, HUES7
Fixative Paraformaldehyde
Permeabilization Yes - Triton
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Username

Ms. Fiona Lewis

Verified customer

Submitted Aug 05 2009

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (embryonic stem cells)
Specification embryonic stem cells
Fixative Paraformaldehyde
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 5%
Username

Dr. Maria Keramari

Verified customer

Submitted Dec 06 2006

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