Product Information

Contents: Allophycocyanin-Alexa Fluor® 750 (APC-Alexa Fluor® 750) anti-mouse CD4 (L3T4)
Catalog Number: 27-0042
Formulation: Phosphate buffer pH 7.2,
500 mM NaCl, 0.09% NaN₃, 0.2% BSA, 0.1% gelatin
Storage Conditions: Store at 4°C.
DO NOT FREEZE.
LIGHT-SENSITIVE MATERIAL.
This tandem dye is sensitive to photo-induced oxidation. Protect this vial from light during storage, handling & experimental procedures. This product is guaranteed for 6 months upon receipt when stored properly.
Clone: RM4-5
Isotype: Rat IgG2a, κ

Staining of C57Bl/6 splenocytes with APC anti-mouse CD3e (145-2C11) (cat. 17-0031) and 0.125 μg of APC-Alexa Fluor® 750 Rat IgG2a Iso Cntrl (cat. 27-4321) (left) or 0.125 μg of APC-Alexa Fluor® 750 anti-mouse CD4 (RM4-5) (right). Cells in the lymphocyte gate were used for analysis.

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
14-0042	Affinity Purified anti-mouse CD4 (L3T4)	N/A	N/A	FA FC IH/F
16-0042	Functional Grade* Purified anti-mouse CD4 (L3T4)	N/A	N/A	FC
13-0042	Biotin anti-mouse CD4 (L3T4)	N/A	N/A	FC
11-0042	FITC anti-mouse CD4 (L3T4)	488	518	FC
12-0042	PE anti-mouse CD4 (L3T4)	488	575	FC
15-0042	PE-Cy5 anti-mouse CD4 (L3T4)	488	670	FC
17-0042	APC anti-mouse CD4 (L3T4)	633	660	FC
25-0042	PE-Cy7 anti-mouse CD4 (L3T4)	488	760	FC
56-0042	Alexa Fluor® 700 anti-mouse CD4 (L3T4) (APC-Cy5.5 Equivalent)	633	723	FC
45-0042	PerCP-Cy5.5 anti-mouse CD4 (L3T4) (PE-Cy5.5 replacement)	488	690	FC
47-0042	Allophycocyanin-eFluor^TM 780 anti-mouse CD4 (L3T4) (APC-Alexa Fluor® 750 replacement)	633	780	FC
48-0042	eFluor^TM 450 anti-mouse CD4 (L3T4) (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 RM4-5 monoclonal antibody reacts with the mouse CD4 molecule, a 55 kDa cell surface receptor expressed by a majority of thymocytes, subpopulation of mature T cells and dendritic cells. CD4 binds to MHC class II on the surface of antigen presenting cells and plays an important role both in T cell development and in optimal functioning of mature T cells. In T cells, CD4 associates with protein tyrosine kinase p56lck through its cytoplasmic tail. Binding of RM4-5 is blocked by GK1.5.

Applications Reported

For research use only, not for diagnostic or therapeutic use. This RM4-5 antibody has been reported for use in flow cytometric analysis.

The photo-emission properties and compensation required for Alexa Fluor® 750 are similar to APC-Cy7; however Alexa Fluor® 750 has several advantages when compared to APC-Cy7. The Alexa Fluor® 750 has peak emission of 779 nm, and usually has a slightly higher mean fluorescent intensity when compared to the same clone conjugated with APC-Cy7, which has peak emission of 767 nm. Furthermore, Alexa Fluor® 750 is considered to be a more photostable fluorochrome as compared with APC-Cy7. It is advised not to use fixatives with Alexa Fluor® 750 as this may lead to degradation of the fluorochrome and loss of staining.

Applications Tested

This RM4-5 antibody has been tested by flow cytometric analysis of mouse splenocytes. This can be used at less than or equal to 0.25 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

Bliss SK, Bliss SP, Beiting DP, Alcaraz A, Appleton JA. IL-10 regulates movement of intestinally derived CD4+ T cells to the liver. J Immunol. 2007 Jun 15;178(12):7974-83. (RM4-5, FC, PubMed)

Rubinstein MP, Kovar M, Purton JF, Cho JH, Boyman O, Surh CD, Sprent J. Converting IL-15 to a superagonist by binding to soluble IL-15R{alpha}. Proc Natl Acad Sci U S A. 2006 Jun 13;103(24):9166-71. (RM4-5, FC, PubMed)

Irie J, Wu Y, Wicker LS, Rainbow D, Nalesnik MA, Hirsch R, Peterson LB, Leung PS, Cheng C, Mackay IR, Gershwin ME, Ridgway WM. NOD.c3c4 congenic mice develop autoimmune biliary disease that serologically and pathogenetically models human primary biliary cirrhosis. J Exp Med. 2006 May 15;203(5):1209-19. (RM4-5, IHC frozen, PubMed)

Andres PG, Beck PL, Mizoguchi E, Mizoguchi A, Bhan AK, Dawson T, Kuziel WA, Maeda N, MacDermott RP, Podolsky DK, Reinecker HC. Mice with a selective deletion of the CC chemokine receptors 5 or 2 are protected from dextran sodium sulfate-mediated colitis: lack of CC chemokine receptor 5 expression results in a NK1.1+ lymphocyte-associated Th2-type immune response in the intestine. J Immunol. 2000 Jun 15;164(12):6303-12. (RM4-5, IHC frozen)

Okumura, K. 2000.Personal communication.

Shi Y, Kaliyaperumal A, Lu L, Southwood S, Sette A, Michaels MA, Datta SK. Promiscuous presentation and recognition of nucleosomal autoepitopes in lupus: role of autoimmune T cell receptor alpha chain. J Exp Med. 1998 Feb 2;187(3):367-78. (RM4-5, FA)

Wilde DB, Marrack P, Kappler J, Dialynas DP, Fitch FW. Evidence implicating L3T4 in class II MHC antigen reactivity; monoclonal antibody GK1.5 (anti-L3T4a) blocks class II MHC antigen-specific proliferation, release of lymphokines, and binding by cloned murine helper T lymphocyte lines. J Immunol. 1983 Nov;131(5):2178-83.

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