Thy1 (Thy1.2) Mouse Monoclonal Antibody [Clone ID: 5a-8]

CAT#: CL039B

Thy1 (Thy1.2) mouse monoclonal antibody, clone 5a-8, Biotin


USD 230.00

2 Weeks*

Size
    • 100 ug

Product Images

Specifications

Product Data
Clone Name 5a-8
Applications FC, IHC
Recommended Dilution Flow Cytometry (protocol see below).
Appropriate control samples should always be included in any labelling studies.
Reactivities Mouse
Host Mouse
Isotype IgG2b
Clonality Monoclonal
Immunogen CBA/J
Donor: AKR/J Spleen
Specificity This antibody detects CD90 (Thy 1.2).
It reacts with all T lymphocytes from mouse strains expressing the Thy 1.2 phenotype (i.e. C57BL/6, C3H/He, DBA/2, CBA/J, BALB/c), but does not react with lymphocytes expressing the Thy 1.1 phenotype (i.e. AKR/J, B6.PL (74 NS).
Formulation PBS containing 0.02% Sodium Azide and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml.
Label: Biotin
State: Liquid purified Ig fraction.
Concentration 0.1 mg/ml
Purification Protein G Chromatography.
Conjugation Biotin
Background CD90 / Thy1 antigen is a GPI linked glycoprotein member of the Immunoglobulin superfamily. It is expressed on murine T cells, thymocytes, neural cells, cells of granulocytic lineage, early hematopoietic progenitors, fibroblasts, neurons and Kupffer's cells. Thy1 may play a role in cell to cell or cell to ligand interactions during synaptogenesis and other events in the brain. It is found in most mouse strains except AKR/J, A, Thy1.1 and B6.PL (74NS) expressing Thy1.1.
Synonyms Thy-1, THY1, CDw90
Note Protocol: FLOW CYTOMETRY ANALYSIS:

Method:

1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cell population.
2. Wash 2 times.
3. Resuspend the cells to a concentration of 2x10e7 cells/ml in media A. Add 50 µl of this suspension to each tube (each tube will then contain 1 x 10e6 cells, representing 1 test).
4. To each tube, add 0.2-0.5 µg of this antibody per 10e6 cells.
5. Vortex the tubes to ensure thorough mixing of antibody and cells.
6. Incubate the tubes for 30 minutes at 4°C.
7. Wash 2 times at 4°C.
8. Add 100 µl of secondary antibody (Streptavidin-FITC) at a 1/500 dilution.
9. Incubate tubes at 4°C for 30-60 minutes (It is recommended that tubes are protected from light since most fluorochromes are light sensitive).
10. Wash 2 times at 4°C.
11. Resuspend the cell pellet in 50 µl ice cold media B.
12. Transfer to suitable tubes for flow cytometric analysis containing 15 µl of propidium iodide at 0.5 mg/ml in PBS. This stains dead cells by intercalating in DNA.

Media:
A. Phosphate buffered saline (pH 7.2) + 5% normal serum of host species + sodium azide (100 µl of 2M sodium azide in 100 mls).
B. Phosphate buffered saline (pH 7.2) + 0.5% Bovine serum albumin + sodium azide (100 µl of 2M sodium azide in 100 mls).

Results:
Tissue Distribution by Flow Cytometry Analysis:
Mouse Strain: CBA/J
Cell Concentration : 1x10e6 cells per tests
Antibody Concentration Used: 0.2 µg/10e6 cells
Isotypic Control: Biotin Mouse IgG2b,k
Cell Source: Percentage of cells stained above control:
Thymus: 97.8%
Spleen: 35.4%
Reference Data

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*Delivery time may vary from web posted schedule. Occasional delays may occur due to unforeseen complexities in the preparation of your product. International customers may expect an additional 1-2 weeks in shipping.