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Alexa Fluor® 647 anti-mouse CD4 Antibody

Pricing & Availability
Clone
RM4-5 (See other available formats)
Regulatory Status
RUO
Other Names
L3T4, T4
Isotype
Rat IgG2a, κ
Ave. Rating
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Product Citations
publications
1_RM4-5_Alx647_030206
C57BL/6 mouse splenocytes stained with CD4 (clone RM4-5) Alexa Fluor® 647 (filled histogram) or rat IgG2a, κ Alexa Fluor® 647 isotype control (open histogram).
  • 1_RM4-5_Alx647_030206
    C57BL/6 mouse splenocytes stained with CD4 (clone RM4-5) Alexa Fluor® 647 (filled histogram) or rat IgG2a, κ Alexa Fluor® 647 isotype control (open histogram).
  • 2_Still-image_0401-5_10X-1_1
    Formalin-fixed, 300 micron-thick mouse spleen section was blocked, permeabilized and stained overnight with CD4 (clone RM4-5) Alexa Fluor® 647 (red), CD169 (Siglec-1)(clone 3D6.112) Alexa Fluor® 488 (green), and CD45R/B220 (clone RA3-6B2) Alexa Fluor® 594 (blue) all at 5 µg/mL, optically cleared, then analyzed at 220 μm imaging depth on a confocal microscope. Watch the video.
  • 3-RM4-5_A647_CD4_Antibody_IHCF_060719
    C57BL/6 mouse frozen spleen section was fixed with 4% paraformaldehyde (PFA) for 10 minutes at room temperature and blocked with 5% FBS for 30 minutes at room temperature. Then the section was stained with 10 µg/mL of anti-mouse CD4 (clone RM4-5) Alexa Fluor® 647 (red), anti-mouse CD8b (clone YTS156.7.7) Brilliant Violet 421™ (blue) and anti-mouse B220 (clone RA3-6B2) Alexa Fluor® 488 (green) overnight at 4°C. The image was captured by 10X objective.
  • RM4-5_A647_CD4_Antibody_3D_IHC_05112021.png
    Paraformaldehyde-fixed (4%), 500 μm-thick mouse spleen tissue section was processed according to the Ce3DTM Tissue Clearing Kit protocol (cat. no. 427701). The section was costained with anti-mouse/human CD45R/B220 Antibody (clone RA3-6B2) Alexa Fluor® 488 at 5 µg/mL (green), anti-mouse CD18 Antibody (clone M18/2) Alexa Fluor® 594 at 5 µg/mL (yellow), and anti-mouse CD4 Antibody (clone RM4-5) Alexa Fluor® 647 at 5 µg/mL (magenta). The section was then optically cleared and mounted in a sample chamber. The image was captured with a 10X objective using Zeiss 780 confocal microscope and processed by Imaris image analysis software.
    Watch the video.
Compare all formats See Alexa Fluor® 647 spectral data
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100533 25 µg 81€
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100530 100 µg 187€
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Description

CD4 is a 55 kD protein also known as L3T4 or T4. It is a member of the Ig superfamily, primarily expressed on most thymocytes and a subset of T cells, and weakly on macrophages and dendritic cells. It acts as a co-receptor with the TCR during T cell activation and thymic differentiation by binding MHC class II and associating with the protein tyrosine kinase lck.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Mouse
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
BALB/c mouse thymocytes
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 647 under optimal conditions.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application

FC - Quality tested
3D IHC, IHC-F - Verified
Recommended Usage

Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is ≤ 0.25 µg per 106 cells in 100 µL volume. For 3D immunohistochemistry on formalin-fixed tissues, a concentration of 5.0 µg/mL is suggested. For immunohistochemistry on frozen tissue sections, a concentration range of 5.0 - 10 µg/mL is suggested. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 647 has a maximum emission of 668 nm when it is excited at 633nm / 635nm.


Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.

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Excitation Laser
Red Laser (633 nm)
Application Notes

The RM4-5 antibody blocks the binding of GK1.5 antibody and H129.19 antibody to CD4+ T cells, but not RM4-4 antibody. Additional reported applications (for the relevant formats) include: blocking of ligand binding, in vivo depletion of CD4+ cells1, and immunohistochemistry of acetone-fixed frozen tissue sections2,3,11 and paraffin-embedded sections11. Clone RM4-5 is not recommended for immunohistochemistry of formalin-fixed paraffin sections. Instead, acetone frozen or zinc-fixed paraffin sections are recommended. The Ultra-LEAF™ Purified antibody (Endotoxin < 0.01 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays (Cat. No. 100575 and 100576).

Application References

(PubMed link indicates BioLegend citation)
  1. Kruisbeek AM. 1991. In Curr. Protocols Immunol. pp. 4.1.1-4.1.5. (Block, Deplete)
  2. Nitta H, et al. 1997. Cell Vision 4:73. (IHC)
  3. Fan WY, et al. 2001. Exp. Biol. Med. 226:1045.
  4. Muraille E, et al. 2003. Infect. Immun. 71:2704. (IHC)
  5. León-Ponte M, et al. 2007. Blood 109:3139. (FC)
  6. Bourdeau A, et al. 2007. Blood doi:10.1182/blood-2006-08-044370. (FC)
  7. Matsumoto M, et al. 2007.J. Immunol.178:2499. PubMed
  8. Shigeta A, et al. 2008. Blood 112:4915. PubMed
  9. Zaborsky N, et al. 2010. J. Immunol. 184:725. PubMed
  10. Rodrigues-Manzanet R, et al. 2010. P. Natl Acad Sci USA 107:8706. PubMed
  11. Whiteland JL, et al. 1995. J. Histochem. Cytochem. 43:313. (IHC)
Product Citations
  1. Ozga AJ, et al. 2022. Immunity. 55:82. PubMed
  2. Cautivo KM, et al. 2022. Immunity. 55:254. PubMed
  3. El-Naccache DW, et al. 2022. Cell Rep. 40:111150. PubMed
  4. Yeh CH, et al. 2022. Immunity. 55:272. PubMed
  5. Hu HJ, et al. 2020. Cell Death Dis. 1.168055556. PubMed
  6. Matsumoto R, et al. 2021. Front Pharmacol. 12:715752. PubMed
  7. Yang W, et al. 2020. Mucosal Immunol. 13:788. PubMed
  8. Liu M, et al. 2020. Nature. 587:115. PubMed
  9. Montel-Hagen A, et al. 2020. Cell Rep. 33:108320. PubMed
  10. Dikiy S, et al. 2021. Immunity. 54(5):931-946.e11. PubMed
  11. Zhou JY, et al. 2021. eLife. 10:00. PubMed
  12. Ng SS, et al. 2020. Nat Immunol. 21:1205. PubMed
  13. Sinclair LV et al. 2019. Elife. 8 pii: e44210. PubMed
  14. Jain RW, et al. 2018. Cell Rep. 25:3342. PubMed
  15. Collins N, et al. 2016. Nat Commun. 7:11514. PubMed
  16. Jamali A, et al. 2020. Cell Reports. 32(9):108099. PubMed
  17. Beloor J et al. 2018. Cell host & microbe. 23(4):549-556 . PubMed
  18. Gomez S, et al. 2022. J Immunother Cancer. 10:. PubMed
  19. Niderla-Bielinska J, et al. 2021. Int J Mol Sci. :22. PubMed
  20. Thornton T, et al. 2016. Nat Commun. 7:10553. PubMed
  21. Rouleau N, et al. 2020. Immunohorizons. 0.695833333. PubMed
  22. Miyauchi K, et al. 2021. Nat Commun. 12:3789. PubMed
  23. Freed-Pastor WA, et al. 2021. Cancer Cell. 39:1342. PubMed
  24. Chryplewicz A, et al. 2022. Cancer Cell. 40:1111. PubMed
  25. Katsuyama T, et al. 2019. J Clin Invest. 129:5411. PubMed
  26. Kim SI, et al. 2020. Molecular Cancer Therapeutics. 20(1):173-182. PubMed
RRID
AB_493372 (BioLegend Cat. No. 100533)
AB_493372 (BioLegend Cat. No. 100530)

Antigen Details

Structure
Ig superfamily, 55 kD
Distribution

Majority of thymocytes, T cell subset

Function
TCR co-receptor, T cell activation
Ligand/Receptor
MHC class II molecule
Cell Type
Dendritic cells, T cells, Thymocytes, Tregs
Biology Area
Immunology
Molecular Family
CD Molecules
Antigen References

1. Barclay A, et al. 1997. The Leukocyte Antigen FactsBook Academic Press.
2. Bierer BE, et al. 1989. Annu. Rev. Immunol. 7:579.
3. Janeway CA. 1992. Annu. Rev. Immunol. 10:645.

Gene ID
12504 View all products for this Gene ID
UniProt
View information about CD4 on UniProt.org

Related FAQs

I am unable to see expression of T cell markers such as CD3 and CD4 post activation.
TCR-CD3 complexes on the T-lymphocyte surface are rapidly downregulated upon activation with peptide-MHC complex, superantigen or cross-linking with anti-TCR or anti-CD3 antibodies. PMA/Ionomycin treatment has been shown to downregulate surface CD4 expression. Receptor downregulation is a common biological phenomenon and so make sure that your stimulation treatment is not causing it in your sample type.
Go To Top Version: 8    Revision Date: 05.11.2021

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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