MHC Class I H-2Ld Mouse Monoclonal Antibody [Clone ID: 30-5-7S]

CAT#: CL060B

MHC Class I H-2Ld mouse monoclonal antibody, clone 30-5-7S, Biotin

Conjugation: Biotin FITC



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CNY 3,710.00


货期*
5周

规格
    • 100 ug

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Specifications

Product Data
Clone Name 30-5-7S
Applications FC
Recommend Dilution Flow Cytometry (See Protocol below).
Reactivity Mouse
Host Mouse
Clonality Monoclonal
Specificity This antibody antibody detects the public specificity H-2.65 of the H-2Ld antigen.
It also recognizes H-2Dq and H-2Lq molecules.
Formulation PBS containing 0.02% Sodium Azide as preservative and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml.
Label: Biotin
State: Liquid purrified Ig fraction.
Concentration lot specific
Purification Protein G Chromatography.
Conjugation Biotin
Storage Condition Store the antibody undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer.
Avoid repeated freezing and thawing.
Background The classical MHC Class I molecules are histocompatibility antigens encoded by the H-2 gene complex and consist of heterodimers of highly polymorphic alpha chains noncovalently associated with the invariant Beta2-microglobulin. These antigens are expressed on most nucleated cells but expression varies on different cell types. MHC Class I molecules present endogenously synthesized peptides to CD8+ T lymphocytes, which are usually cytotoxic T cells. MHC Class I antigens expressed on thymic epithelial cells regulate the positive and negative selection of CD8+ T cells during T cell ontogeny.
Synonyms H2-L
Note Protocol: FLOW CYTOMETRY ANALYSIS:
Method:
1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cell population with cell separation medium.
2. Wash 2 times.
3. Resuspend the cells to a concentration of 2x10e7cells/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 1.0 µg of 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: Balb/c
Cell Concentration : 1x10e6 cells per test
Antibody Concentration Used: 1.0 µg/10e6 cells
Isotypic Control: Biotin Mouse IgG2a

Cell Source Percentage of cells stained above control: (See Figure 1.)
Spleen 91.1%
Thymus 99.4%
Lymph Node 98.1%

Strain Distribution:
Strains Tested:
Strain Haplotype + / -
BALB/c H-2d +
A.TH H-2KsDd +
A.TL H-2KsDd +
B.10A(3R) H-2KbDd +
C3H/He H-2k -
C57BL/6 H-2b -
Reference Data
*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.
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