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FITC anti-human CD4 Antibody

Pricing & Availability
Clone
RPA-T4 (See other available formats)
Regulatory Status
RUO
Workshop
IV T114
Other Names
T4
Isotype
Mouse IgG1, κ
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Product Citations
publications
1_RPA-T4_FITC_120108.jpg
Human peripheral blood lymphocytes stained with RPA-T4 FITC
  • 1_RPA-T4_FITC_120108.jpg
    Human peripheral blood lymphocytes stained with RPA-T4 FITC
  • 2_11_Human_LN_CD163_CD4
    Confocal image of human lymph node sample acquired using the IBEX method of highly multiplexed antibody-based imaging: CD163 (red) in Cycle 3 and CD4 (blue) in Cycle 5. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
See FITC spectral data
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300505 25 tests 14€
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300506 100 tests 40€
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300538 500 tests 172€
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Description

CD4, also known as T4, is a 55 kD single-chain type I transmembrane glycoprotein expressed on most thymocytes, a subset of T cells, and monocytes/macrophages. CD4, a member of the Ig superfamily, recognizes antigens associated with MHC class II molecules, and participates in cell-cell interactions, thymic differentiation, and signal transduction. CD4 acts as a primary receptor for HIV, binding to HIV gp120. CD4 has also been shown to interact with IL-16.

Product Details
Technical Data Sheet (pdf)

Product Details

Reactivity
Human
Antibody Type
Monoclonal
Host Species
Mouse
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and BSA (origin USA)
Preparation
The antibody was purified by affinity chromatography, and conjugated with FITC under optimal conditions.
Concentration
Lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.)
Storage & Handling
The CD4 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
SB - Reported in the literature, not verified in house
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 5 µl per million cells in 100 µl staining volume or 5 µl per 100 µl of whole blood.

Excitation Laser
Blue Laser (488 nm)
Application Notes

The RPA-T4 antibody binds to the D1 domain of CD4 (CDR1 and CDR3 epitopes) and can block HIV gp120 binding and inhibit syncytia formation. Additional reported applications (for the relevant formats) include: immunohistochemistry of acetone-fixed frozen sections3,4,5, blocking of T cell activation1,2, and spatial biology (IBEX)10,11.  This clone was tested in-house and does not work on formalin fixed paraffin-embedded (FFPE) tissue. The Ultra-LEAF™ purified antibody (Endotoxin < 0.01 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays (Cat. No. 300569 - 300574).

Additional Product Notes

Iterative Bleaching Extended multi-pleXity (IBEX) is a fluorescent imaging technique capable of highly-multiplexed spatial analysis. The method relies on cyclical bleaching of panels of fluorescent antibodies in order to image and analyze many markers over multiple cycles of staining, imaging, and, bleaching. It is a community-developed open-access method developed by the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).

Application References
  1. Knapp W, et al. 1989. Leucocyte Typing IV. Oxford University Press. New York. (Activ)
  2. Moir S, et al. 1999. J. Virol. 73:7972. (Activ)
  3. Deng MC, et al. 1995. Circulation 91:1647. (IHC)
  4. Friedman T, et al. 1999. J. Immunol. 162:5256. (IHC)
  5. Mack CL, et al. 2004. Pediatr. Res. 56:79. (IHC)
  6. Lan RY, et al. 2006. Hepatology 43:729.
  7. Zenaro E, et al. 2009. J. Leukoc. Biol. 86:1393. (FC) PubMed
  8. Yoshino N, et al. 2000. Exp. Anim. (Tokyo) 49:97. (FC)
  9. Stoeckius M, et al. 2017. Nat. Methods. 14:865. (PG)
  10. Radtke AJ, et al. 2020. Proc Natl Acad Sci USA. 117:33455-33465. (SB) PubMed
  11. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
  1. Meng Y,et al. 2017. Cell Death Dis. . 10.1038/cddis.2017.505. PubMed
  2. Goletz C, et al. 2018. Front Immunol. 9:1614. PubMed
  3. Sibener LV et al. 2018. Cell. 174(3):672-687 . PubMed
  4. Ickrath P, et al. 2019. Biomed Rep. 10:119. PubMed
  5. Iio K, et al. 2019. Sci Rep. 9:813. PubMed
  6. Alhaj Hussen K, et al. 2017. Immunity. 47:680. PubMed
  7. Ickrath P, et al. 2018. Int J Mol Med. 42:1116. PubMed
  8. Hansen EC et al. 2016. eLife. 5 pii: e18447. PubMed
  9. Zhao Y, et al. 2020. Front Immunol. 2.572222222. PubMed
  10. Kang LJ, et al. 2020. Sci Rep. 10:5603. PubMed
  11. Mothe B, et al. 2020. Front Immunol. 1.029861111. PubMed
  12. Soldi R, et al. 2020. PLoS One. 15:e0235705. PubMed
  13. Handono K, et al. 2020. Eur J Dent. 0.961111111. PubMed
  14. Evans RDR, et al. 2020. Nat Commun. 3.491666667. PubMed
  15. Zenaro E, et al. 2009. J Leukoc Biol. 86:1393. PubMed
  16. Tamai Y, et al. 2013. J Immunol. 190:4382. PubMed
  17. Choudhry V, et al. 2006. Biochem Biophys Res Commun. 348:1107. PubMed
  18. Han L, et al. 2014. J Biol Chem. 289:25546. PubMed
  19. Luo X, et al. 2015. J Biol Chem. 290: 28675 - 28682. PubMed
  20. Wahl S, et al. 2016. Nature. 541:81-86. PubMed
  21. Rolandelli A, et al. 2017. Sci Rep. 7:40666. PubMed
  22. Gong B, et al. 2021. Mol Med Rep. 23:00. PubMed
  23. Vanoni G, et al. 2021. eLife. 10:00. PubMed
  24. Hamilton JR, et al. 2021. Cell Reports. 35(9):109207. PubMed
  25. Cheng Y, et al. 2021. Immunity. 54(8):1825-1840.e7. PubMed
  26. Qiu XM, et al. 2020. Reproduction. 251:159. PubMed
  27. Kobayashi Y, et al. 2020. Int J Oncol. 999:56. PubMed
  28. Zhao J, et al. 2021. Front Immunol. 12:658420. PubMed
  29. Panwar B, et al. 2021. Genome Res. 31:659. PubMed
  30. Scherpenisse M, et al. 2021. MBio. 12:. PubMed
  31. Chiu H, et al. 2021. iScience. 24:102748. PubMed
  32. Fang F, et al. 2021. Cell Rep. 37:109981. PubMed
  33. Yang L, et al. 2020. Genes Dis. 7:128. PubMed
  34. Pagel J, et al. 2020. Front Immunol. 11:565257. PubMed
  35. Zong D, et al. 2021. BMC Biol. 19:79. PubMed
  36. Kim MY, et al. 2021. JCI Insight. 6:. PubMed
  37. Kim ML, et al. 2021. iScience. 24:103509. PubMed
  38. Beatson RE, et al. 2021. Cell Rep Med. 2:100473. PubMed
  39. Le X, et al. 2021. J Thorac Oncol. 16:583. PubMed
  40. Kim MY, et al. 2022. Nat Commun. 13:3296. PubMed
  41. Fang F, et al. 2022. JCI Insight. 7:. PubMed
  42. Zelba H, et al. 2021. J Immunol. 206:580. PubMed
  43. Kothari H, et al. 2021. Sci Signal. 14:. PubMed
  44. Csomós K, et al. 2022. Nat Immunol. 23:1256. PubMed
  45. Su W, et al. 2022. Front Immunol. 13:952338. PubMed
  46. Kalim H, et al. 2020. Int J Rheum Dis. 23:620. PubMed
  47. Sun L, et al. 2020. J Diabetes Res. 2020:2583257. PubMed
  48. Mastrogiovanni M, et al. 2022. Sci Adv. 8:eabl5942. PubMed
  49. Reitinger C, et al. 2022. Front Immunol. 13:970290. PubMed
  50. Herter JM, et al. 2022. Strahlenther Onkol. Online ahead of print. PubMed
  51. Jiao Q, et al. 2020. Br J Dermatol. 182:648. PubMed
  52. Hagel J, et al. 2021. J Immunol. 206:3073. PubMed
  53. Saber MM, et al. 2022. J Immunol Res. 2022:7219207. PubMed
  54. Gartshteyn Y, et al. 2023. Life Sci Alliance. 6: . PubMed
  55. He T, et al. 2023. Immun Inflamm Dis. 11:e759. PubMed
  56. Iio K, et al. 2023. Immun Ageing. 20:8. PubMed
  57. Yen M, et al. 2022. Cell. 185:1414. PubMed
  58. Yang Y, et al. 2023. MBio. 14:e0328522. PubMed
  59. Mohammad TAM, et al. 2023. Int J Rheum Dis. 26:740. PubMed
  60. Li J, et al. 2023. Sci Rep. 13:7501. PubMed
RRID
AB_314073 (BioLegend Cat. No. 300505)
AB_314074 (BioLegend Cat. No. 300506)
AB_2562052 (BioLegend Cat. No. 300538)

Antigen Details

Structure
Ig superfamily, type I transmembrane glycoprotein, 55 kD
Distribution

T cell subset, majority of thymocytes, monocytes/macrophages

Function
MHC class II co-receptor, lymphocyte adhesion, thymic differentiation, HIV receptor
Ligand/Receptor
MHC class II molecules, HIV gp120, IL-16
Cell Type
Dendritic cells, Macrophages, Monocytes, T cells, Thymocytes, Tregs
Biology Area
Immunology
Molecular Family
CD Molecules
Antigen References

1. Center D, et al. 1996. Immunol. Today 17:476.
2. Gaubin M, et al. 1996. Eur. J. Clin. Chem. Clin. Biochem. 34:723.

Gene ID
920 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.
If an antibody clone has been previously successfully used in IBEX in one fluorescent format, will other antibody formats work as well?

It’s likely that other fluorophore conjugates to the same antibody clone will also be compatible with IBEX using the same sample fixation procedure. Ultimately a directly conjugated antibody’s utility in fluorescent imaging and IBEX may be specific to the sample and microscope being used in the experiment. Some antibody clone conjugates may perform better than others due to performance differences in non-specific binding, fluorophore brightness, and other biochemical properties unique to that conjugate.

Will antibodies my lab is already using for fluorescent or chromogenic IHC work in IBEX?

Fundamentally, IBEX as a technique that works much in the same way as single antibody panels or single marker IF/IHC. If you’re already successfully using an antibody clone on a sample of interest, it is likely that clone will have utility in IBEX. It is expected some optimization and testing of different antibody fluorophore conjugates will be required to find a suitable format; however, legacy microscopy techniques like chromogenic IHC on fixed or frozen tissue is an excellent place to start looking for useful antibodies.

Are other fluorophores compatible with IBEX?

Over 18 fluorescent formats have been screened for use in IBEX, however, it is likely that other fluorophores are able to be rapidly bleached in IBEX. If a fluorophore format is already suitable for your imaging platform it can be tested for compatibility in IBEX.

The same antibody works in one tissue type but not another. What is happening?

Differences in tissue properties may impact both the ability of an antibody to bind its target specifically and impact the ability of a specific fluorophore conjugate to overcome the background fluorescent signal in a given tissue. Secondary stains, as well as testing multiple fluorescent conjugates of the same clone, may help to troubleshoot challenging targets or tissues. Using a reference control tissue may also give confidence in the specificity of your staining.

How can I be sure the staining I’m seeing in my tissue is real?

In general, best practices for validating an antibody in traditional chromogenic or fluorescent IHC are applicable to IBEX. Please reference the Nature Methods review on antibody based multiplexed imaging for resources on validating antibodies for IBEX.

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Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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