AM31877BT-N MHC Class II (I-Ab,d) antibody

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0.1 mg / €260.00
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Mouse anti MHC Class II (I-Ab,d) 25-9-17S

AM31877BT-N

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  • AM31877BT-N

Product Description for MHC Class II (I-Ab,d)

Mouse anti MHC Class II (I-Ab,d) 25-9-17S.
Presentation: Biotin
Product is tested for Flow Cytometry.

Properties for MHC Class II (I-Ab,d)

Product Category Primary Antibodies
Quantity 0.1 mg
Presentation Biotin
Applications F
Clonality Monoclonal
Clone 25-9-17S
Host Mouse
Isotype IgG2a
Shipping to Worldwide
PDF datasheet View Datasheet
Manufacturer Acris Antibodies GmbH
Material safety datasheet MSDS for Monoclonal Antibodies (de)

Datasheet Extract

Immunogen
Immunogen:
C3H.SW splenocytes.
Donor: C3H lymphoid cells.
Fusion Partner: Sp2/0-Ag14
Application Flow Cytometry (See Protocols).
Concentration 0.1 mg/ml
Protocols FLOW CYTOMETRY ANALYSIS:
Method:
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 106 cells, representing 1 test).
4. To each tube, add 0.5-0.2 μg* of AM31877BT-N 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: C57BL/6
Cell Concentration : 1x10e6 cells per test.
Antibody Concentration Used: 0.5 μg/106 cells.
Isotypic Control: Biotin Mouse IgG2a

Cell Source Percentage of cells stained above control:
Thymus: 17.5%
Spleen: 58.6%
Lymph Node: 31.2%

Strain Distribution by Flow Cytometry Analysis:
Antibody Concentration: 0.5 µg/106 cells.
Strains Tested: See Figure 2.
General Readings
  1. Ozato K, Sachs DH. Monoclonal antibodies to mouse MHC antigens. III. Hybridoma antibodies reacting to antigens of the H-2b haplotype reveal genetic control of isotype expression. J Immunol. 1981 Jan;126(1):317-21. PubMed PMID: 6935293.
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.
Shelf life: one year from despatch.
Format
Label:
Biotin
Purification:
Protein G Chromatography
Buffer System:
PBS containing 0.02% Sodium Azideas preservative and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml
State:
Liquid purified Ig fraction
Specificity
Specificity:
This Monoclonal Antibody reacts with I-Ab and I-Ad antigens. Cross reaction with H-2p and H-2q was also found.
Species:
Mouse.

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