LOC102641613 Mouse Monoclonal Antibody [Clone ID: 34-5-8S]

CAT#: CL053B

LOC102641613 mouse monoclonal antibody, clone 34-5-8S, Biotin

Conjugation: Biotin FITC

USD 371.00

2 Weeks*

    • 100 ug

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Product Data
Clone Name 34-5-8S
Applications FC
Recommended Dilution Flow cytometry.
Reactivities Mouse
Host Mouse
Isotype IgG2a
Clonality Monoclonal
Immunogen B6xDBA/2 spleen cells
Donor: C3H/HeJ spleen
Fusion Partner: SP2/0.Ag14
Specificity This mAb is specific for cells expressing the H-2D antigen coded for by the d haplotype. The reaction pattern of this antibody with a panel of inbred and recombinant haplotypes demonstrates that the antibody detects a private determinant (H-2.4) of the H-2Dd antigen. This antibody can be used to quantitate cells bearing H-2Dd (H-2.4) antigen from the appropriate strains of mice.
Formulation PBS , 0.02% NaN3 and EIA grade BSA as a stabalizing protein to bring total protein concentration to 4-5 mg/ml.
Label: Biotin
State: Liquid purified Ig
Concentration lot specific
Purification Protein G Chromatography
Conjugation Biotin
Storage Store the antibody undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer.
Avoid repeated freezing and thawing.
Stability Shelf life: one year from despatch.
Gene Name H-2 class I histocompatibility antigen, D-D alpha chain
Synonyms H2-D1, H-2D(D)

1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cell population with Lympholyte®-M cell separation medium.
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 0.5-1.0 µg* of this Ab 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.

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 tests
Antibody Concentration Used: 1.0 µg/10e6 cells
Isotypic Control: Biotin Mouse IgG2a

Cell Source Percentage of cells stained above control:
Thymus: 40.7 %
Spleen: 98.3%

Strain Distribution by Flow Cytometry Analysis:
Cell Concentration : 1x10e6 cells per tests
Antibody Concentration Used: 1.0 µg/10e6 cells
Strains Tested: BALB/c, C57BL/6, CBA/J, C3H/He
Positive: BALB/c
Negative: C57BL/6, CBA/J, C3H/He
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.