Product Information

Contents: Alexa Fluor® 700 anti-mouse MHC Class II (I-A/I-E)
Catalog Number: 56-5321
Sizes: 25 ug, 100 ug
Formulation: Phosphate buffer pH 7.2,
150 mM NaCl, 0.09% NaN₃, 0.1% gelatin
Storage Conditions: Store at 4°C.
DO NOT FREEZE.
LIGHT-SENSITIVE MATERIAL.
Clone: M5/114.15.2
Isotype: Rat IgG2b, κ

Staining of C57Bl/6 splenocytes with FITC anti-mouse CD3e (145-2C11) (cat. 11-0031) and staining buffer (autofluorescence) (left) or 0.03 μg of Alexa Fluor® 700 anti-mouse Class II (M5/114.15.2) (right). Cells in the lymphocyte gate were used for analysis.

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
14-5321	Affinity Purified anti-mouse MHC Class II (I-A/I-E)	N/A	N/A	FA FC IH/F IHC(Paraffin) IP WB
16-5321	Functional Grade* Purified anti-mouse MHC Class II (I-A/I-E)	N/A	N/A	FC
13-5321	Biotin anti-mouse MHC Class II (I-A/I-E)	N/A	N/A	FC
11-5321	FITC anti-mouse MHC Class II (I-A/I-E)	488	518	FC
12-5321	PE anti-mouse MHC Class II (I-A/I-E)	488	575	FC
15-5321	PE-Cy5 anti-mouse MHC Class II (I-A/I-E)	488	670	FC
17-5321	APC anti-mouse MHC Class II (I-A/I-E)	633	660	FC
36-5321	Functional Grade* Biotin anti-mouse MHC Class II (I-A/I-E)	N/A	N/A	FC IHC(Paraffin) IP
56-5321	Alexa Fluor® 700 anti-mouse MHC Class II (I-A/I-E) (APC-Cy5.5 Equivalent)	633	723	FC
95-5321	eFluor^TM 650^NC anti-mouse MHC Class II (I-A/I-E)	350-500	650	FC
47-5321	Allophycocyanin-eFluor^TM 780 anti-mouse MHC Class II (I-A/I-E) (APC-Alexa Fluor® 750 Replacement)	633	780	FC
48-5321	eFluor^TM 450 anti-mouse MHC Class II (I-A/I-E) (Pacific Blue® replacement)	405	450	FC
*Functional Grade™ (FG™) Purified: Azide-free, sterile-filtered, and endotoxin < 0.001 ng/µg (unless otherwise noted). *Functional Grade™ (FG™) Biotin: Azide-free, sterile-filtered, and endotoxin < 0.05 ng/µg. Purified: Contains azide, not sterile-filtered, and not endotoxin tested.
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Pacific Blue® and APC-Alexa Fluor® 750 conjugated products with eFluor™ 450 and APC-eFluor™ 780 conjugated products, respectively.

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Description

The M5/114.15.2 monoclonal antibody reacts with the mouse major histocompatibility complex class II, both I-A and I-E subregion-encoded glycoproteins (I-A^b, I-A^d, I-A^q, I-E^d, I-E^k, not I-A^f, I-A^k, or I-A^s). It detects a polymorphic determinant present on B cells, monocytes, macrophages, dendritic cells, and activated T lymphocytes from mice carrying the H-2^b, H-2^d, H-2^q, H-2^p, H-2^r and H-2^u but not from mice carrying the H-2^s or H-2^f haplotypes. The M5/114 mAb is reported to inhibit I-A-restricted T cell responses of the H-2^b, H-2^d, H-2^q, H-2^u but not H-2^f, H-2^k, or H-2^s haplotypes.

Applications Reported

For research use only, not for diagnostic or therapeutic use. This M5/114.15.2 antibody has been reported for use in flow cytometric analysis.

The Alexa Fluor® 700 emits at 723 nm and can be excited with the He-Ne 633 laser. Most instruments will require a 685 LP mirror and 710/20 filter. Please make sure that your instrument is capable of detecting this fluorochome.

Applications Tested

This M5/114.15.2 antibody has been tested by flow cytometric analysis of mouse splenocytes. This can be used at less than or equal to 0.06 μg per million cells in a 100 μl total staining volume. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

References

Bhattacharya, A., M. E. Dorf, et al. 1981. A shared alloantigenic determinant on Ia antigens encoded by the I-A and I-E subregions: evidence for I region gene duplication. J Immunol 127(6): 2488-95.

Germain, R. N., A. Bhattacharya, et al. 1982. A single monoclonal anti-Ia antibody inhibits antigen-specific T cell proliferation controlled by distinct Ir genes mapping in different H-2 I subregions. J Immunol 128(3): 1409-13.

Gueirard, Pascale et al. 2003."Bordetella bronchiseptica Perists in the Nasal Cavities of Mice and Triggers Early Delivery of Dendritic Cells in the Lymph Nodes Draining the Lower and Upper Respiratory Tract". Infection and Immunity 71(7):4137-4143. (M5/114.15.2, IHC paraffin, PubMed)

Bagavant, Sharp et al. 2002. "Induction and Immunohistology of Autoimmune Ovarian Disease in Cynomolgus Macaques (Macaca fascicularis)". Am J Pathol 160:141-149. (M5/114.15.2, IHC, PubMed)

Li C, Siemasko K, et al. 2002. Cooperative interaction of Ig(alpha) and Ig(beta) of the BCR regulates the kinetics and specificity of antigen targeting. Int Immunol. 14(10):1179-91. (M5/114.15.2, IHC frozen and WB, PubMed)

Unternaehrer JJ, Chow A, et al. 2007. The tetraspanin CD9 mediates lateral association of MHC class II molecules on the dendritic cell surface. Proc Natl Acad Sci U S A. 104(1):234-9. (M5/114.15.2, IP, PubMed)

Mediratta SK, Singh N, et al. 1996. Analysis of T-cell hybridomas with an unusual MHC class II-dependent ligand specificity. Immunology. 89(2):238-44. (M5/114.15.2, FA, PubMed)

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Alexa Fluor® and Pacific Blue® are registered trademarks of and licensed under patents assigned to Molecular Probes, Inc. for research use only. This product is subject to an agreement between Molecular Probes, Inc. and eBioscience, and the manufacture, use, sale or import of this product may be subject to one or more U.S. patents, pending applications and corresponding foreign equivalents, owned by Molecular Probes, Inc. (a wholly owned subsidiary of Invitrogen Corp). The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product for life science research or as an ASR. The buyer cannot use this product for manufacturing or for any other screening (specifically including use in combination with microarrays or High Content Screening) or testing purpose, other than as an ASR. For information on purchasing a license to this product for purposes other than life science research or use as an ASR, contact Molecular Probes, Inc.