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

CAT#: CL060F

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


USD 250.00

2 Weeks*

Size
    • 100 ug

Product Images

Specifications

Product Data
Clone Name 30-5-7S
Applications Assay, CT, FC
Recommended Dilution Flow Cytometry (see protocol).
Reactivities Mouse
Host Mouse
Isotype IgG2a
Clonality Monoclonal
Immunogen Recipient: BALB/c-H-2dm2
Donor: BALB/c Spleen cells
Fusion Partner: SP2/0.Ag14
Specificity This monoclonal antibody detects the public specificity H-2.65 of the H-2Ld antigen.
This antibody also recognizes H-2Dq and H-2Lq molecules.
Formulation PBS containing 0.02% Sodium Azide as preservative and EIA grade BSA as stabilizing protein to bring the protein concentration to 4-5 mg/ml
Label: FITC
State: Liquid purified IgG fraction
Conjugation FITC
Synonyms H2-L
Note Protocol: Flow Cytometry Analysis:
Method:
1. Prepare cell suspension in Media A. For cell preparations, deplete the red blood cell population with Lympholyte®-M cell separation media.
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 2.0 µg of this Antibody CL060F 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. (It is recommended that tubes are protected from light since most fluorochromes are light sensitive).
7. Wash 2 times at 4°C.
8. Resuspend the cell pellet in 50 µl ice cold Media B.
9. Transfer to suitable tubes for flow cytometric analysis containing 15 µl of propidium iodide at 0.5 mg/ml in phosphate buffered saline. (This stains dead cells by intercalating DNA.)

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

Results in Tissue Distribution by Flow Cytometry Analysis:
Mouse Strain: BALB/c
Cell Concentration: 1x10e6 cells/test
Antibody Concentration Used: 2.0 µg/10e6 cells
Isotype Control: FITC Mouse IgG2a

Cell Source - % of cells stained above Control:
Spleen: 92.4%
Thymus: 54.4%
Lymph Node: 95.3%

Strain Distribution:
Procedure: As above
Antibody Concentration: 1µg/10e6 cells
Strains Tested: See Figure 2
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.