Anti-CD90 / Thy1 antibody [MRC OX-7] (ab225)

Overview

  • Product name
    Anti-CD90 / Thy1 antibody [MRC OX-7]
    See all CD90 / Thy1 primary antibodies
  • Description
    Mouse monoclonal [MRC OX-7] to CD90 / Thy1
  • Specificity
    Recognizes the Thy-1.1 antigenic determinant which is a monomorphic determinant within rat strains but polymorphic in mice. Thus MRC OX-7 reacts with Thy-1.1 mice eg. AKR strain, but not Thy-1.2 mice eg. CBA, BALB/c. As Thy 1 is monomorphic in rat it will react with all rat strains.
  • Tested applications
    Suitable for: Flow Cyt, IP, WB, IHC-P, ICC/IF, IHC-Frmore details
  • Species reactivity
    Reacts with: Rat, Horse
    Predicted to work with: Mouse, Rabbit, Guinea pig
  • Immunogen

    Full length protein corresponding to Rat CD90/ Thy1.

  • Positive control
    • WB: rat brain tissue lysate and PC12 whole cell lysate. IF/ICC: PC12 cells.
  • General notes

    This antibody clone is manufactured by Abcam.

    The affinity of the Fab' of MRC OX-7 for rat Thy-1 is 3 x 109m-1 and for mouse Thy-1.1 is 3 x 108m-1.

     

Properties

Applications

Our Abpromise guarantee covers the use of ab225 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
Flow Cyt Use 0.1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IP Use at an assay dependent concentration.
WB Use a concentration of 5 µg/ml. Detects a band of approximately 35-37 kDa (predicted molecular weight: 17 kDa).
IHC-P Use at an assay dependent concentration. PubMed: 23616767
ICC/IF 1/1000.
IHC-Fr Use at an assay dependent concentration. PubMed: 16723538

Target

  • Function
    May play a role in cell-cell or cell-ligand interactions during synaptogenesis and other events in the brain.
  • Sequence similarities
    Contains 1 Ig-like V-type (immunoglobulin-like) domain.
  • Cellular localization
    Cell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • CD7 antibody
    • CD90 antibody
    • CD90 antigen antibody
    • CDw90 antibody
    • FLJ33325 antibody
    • MGC128895 antibody
    • T25 antibody
    • Theta antigen antibody
    • Thy 1 antibody
    • Thy 1 cell surface antigen antibody
    • Thy 1 membrane glycoprotein antibody
    • Thy 1 T cell antigen antibody
    • Thy 1.2 antibody
    • Thy-1 antigen antibody
    • Thy-1 membrane glycoprotein antibody
    • Thy1 antibody
    • Thy1 antigen antibody
    • Thy1 T cell antigen antibody
    • Thy1.1 antibody
    • Thy1.2 antibody
    • THY1_HUMAN antibody
    • Thymus cell antigen 1, theta antibody
    see all

Images

  • Overlay histogram showing PC12 cells stained with ab225 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab225, 0.1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [B11/6](ab91353, 1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive result in 80% methanol (5 min) fixed PC12 cells used under the same conditions.

  • Immunocytochemistry/ Immunofluorescence analysis of horse adipose and bone marrow stem cells labeling CD90 / Thy1 with ab225 at 1/1000 dilution. Cells were fixed in paraformaldehyde and permeabilized with Triton x100 0.1%. The cells were blocked with 10% BSA for 30 minutes at 37°C, followed by staining with ab225 at 1/1000 for 12 hours in PBS+IGEPAL+BSA+10%NGS at 4°C. Goat F(ab')2 Anti-Mouse IgG - (Fab')2 (Biotin), pre-adsorbed (ab5886) was used as the secondary antibody at 1/400 dilution.

    See Abreview

  • Immunohistochemistry (Frozen sections) analysis of rat spleen tissue labeling CD90 / Thy1 with ab225 at 1/100 dilution. The tissue was fixed in paraformaldehyde  followed by blocking in 10% serum for 20 minutes. The tissue was then stained with ab225 at 1/100 in PBS for 6 hours at 4°C. A polyclonal donkey anti-mouse Alexa Fluor® 488 secondary antibody was used at 1/1000 dilution.

    See Abreview

  • Immunocytochemistry/ Immunofluorescence analysis of rat sciatic nerve schwann cells and fibroblasts labeling CD90 / Thy1 with ab225 at 1/400 dilution. Cells were fixed in paraformaldehyde and permeabilized with 0.5% Triton X-100. The cells were blocked with 5% serum for 1 hour at 21°C, followed by staining with ab225 at 1/400 in 0.2% BSA for 1 hour at 37°C. A polyclonal goat anti-mouse Alexa Fluor® 546 secondary antibody was used at 1/500 dilution.

    See Abreview

  • All lanes : Anti-CD90 / Thy1 antibody [MRC OX-7] (ab225) at 5 µg/ml

    Lane 1 : Brain (Rat) Tissue Lysate
    Lane 2 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 17 kDa
    Observed band size : 35-37 kDa (why is the actual band size different from the predicted?)


    Exposure time : 20 minutes

    Rat CD90/Thy1 is N-glycosylated at three sites, giving rise to molecules with a range of molecular masses (25-37 kDa). 

  • ab225 stained PC12 cells. The cells were 100% methanol fixed for 5 minutes at -20°C and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab225 at 5µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  • This image was kindly supplied as part of the review submitted by Nick Voilley. Rat retinal ganglion cell labelled with ab225 as a primary antibody and an anti-mouse F(ab)' 2 coupled to Alexa488 as a secondary antibody. The cell is approximately 15 micrometers in diameter. Only the cells labelled in green in the culture bear action potentials when stimulated. These three elements (reactivity to ab225, size and elecrophysiological parameters) clearly indicate the cell is a ganglion cell.

References

This product has been referenced in:
  • Du M  et al. Acellular dermal matrix loading with bFGF achieves similar acceleration of bone regeneration to BMP-2 via differential effects on recruitment, proliferation and sustained osteodifferentiation of mesenchymal stem cells. Mater Sci Eng C Mater Biol Appl 70:62-70 (2017). IHC-P ; Rat . Read more (PubMed: 27770935) »
  • Liu J  et al. Homemade-device-induced negative pressure promotes wound healing more efficiently than VSD-induced positive pressure by regulating inflammation, proliferation and remodeling. Int J Mol Med 39:879-888 (2017). ICC/IF ; Rabbit . Read more (PubMed: 28290607) »

See all 34 Publications for this product

Customer reviews and Q&As

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 21°C
Antigen retrieval step
Other
Sample
Rabbit Tissue sections (Bone and cartilage)
Specification
Bone and cartilage
Permeabilization
No
Fixative
70 % Ethanol
Username

Abcam user community

Verified customer

Submitted Aug 26 2014

Application
Flow Cytometry
Sample
Mouse Cell (mMSC)
Specification
mMSC
Preparation
Cell harvesting/tissue preparation method: bone marrow extraction
Sample buffer: PBS
Fixation
Methanol
Permeabilization
No
Username

Abcam user community

Verified customer

Submitted Dec 19 2012

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (mMSC)
Loading amount
70 µg
Specification
mMSC
Gel Running Conditions
Reduced Denaturing (10)
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Dec 18 2012

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Pancreas)
Specification
Pancreas
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate pH 6
Permeabilization
No
Blocking step
FBS / BSA in TBS as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 1% · Temperature: RT°C
Username

Mr. Raul Peña

Verified customer

Submitted Dec 03 2012

Thank you very much for your call yesterday and for your patience.
I have confirmed with the lab that the antibody is tested with non-permeabilized cells in flow cytometry. So, the epitope is extracellular.
I hope that this information will b...

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Thank you for your enquiry and your interest.

Unfortunately, we currently do not have any data (either empirical or from literature) that would support reactivity to porcine tissue. A quick BLAST comparison shows only a 75% homology between t...

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Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry these products did not perform as stated on the datasheet and for the inconvenience th...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Horse Cell (Adipose and bone marrow stem cells)
Specification
Adipose and bone marrow stem cells
Fixative
Paraformaldehyde
Permeabilization
Yes - TRITON x100 0,1%
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 37°C
Username

Ms. Vega Villar

Verified customer

Submitted Mar 29 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (Spleen)
Specification
Spleen
Fixative
Paraformaldehyde
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 10%
Username

Abcam user community

Verified customer

Submitted Sep 06 2011

Application
Immunocytochemistry
Sample
Rat Cultured Cells (Sciatic nerve Schwann cells and fibroblasts)
Specification
Sciatic nerve Schwann cells and fibroblasts
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% Triton X-100
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Oct 08 2008

1-10 of 14 Abreviews or Q&A

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