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Purified anti-human/mouse/rat CD278 (ICOS) Antibody

Pricing & Availability
Clone
C398.4A (See other available formats)
Regulatory Status
RUO
Other Names
Inducible COStimulatory molecule, H4
Isotype
Armenian Hamster IgG
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Product Citations
publications
A_C398_4A_Purified_072007
PHA-stimulated human peripheral blood lymphocytes (3 days) stained with purified C398.4A, followed by biotinylated anti-Armenian hamster IgG and Sav-PE
  • A_C398_4A_Purified_072007
    PHA-stimulated human peripheral blood lymphocytes (3 days) stained with purified C398.4A, followed by biotinylated anti-Armenian hamster IgG and Sav-PE
  • B_C398pt4A_PURE_CD278_Antibody_SB_111623
    SeqIF™ (sequential immunofluorescence) staining on COMET™ of Purified anti-CD278 (ICOS) (clone C398.4A, yellow) on formalin-fixed paraffin-embedded human tonsil tissue at 5 µg/mL. Alexa Fluor® 647 Goat anti-Armenian hamster IgG antibody was used as secondary antibody. Nuclei were counterstained with DAPI (blue). Tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing.
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313502 500 µg 270 CHF
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Description

ICOS, also known as inducible costimulatory molecule and H4, is a 47-57 kD protein. This protein is homologous to the CD28/CTLA-4 proteins. ICOS is expressed on activated T cells and a subset of thymocytes. It is able to costimulate T cells proliferation. In addition, ICOS is involved in humoral immune responses (B cell germinal center formation). The ICOS ligand is B7h/B7RP-1 or B7-H2. ICOS stimulation has been shown to potentiate TCR-mediated IL-4 and IL-10 production and has been proposed to play a role in Th2 cell development.

Product Details
Technical Data Sheet (pdf)

Product Details

Reactivity
Human,Mouse,Rat
Antibody Type
Monoclonal
Host Species
Armenian Hamster
Immunogen
Mouse T cell clone D10.G4.1
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/ml
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

FC - Quality tested
IP, Costim - Reported in the literature, not verified in house
SB - Community 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 ≤ 1.0 µg per 106 cells in 100 µl volume. It is recommended that reagents be titrated for optimal performance in the particular application.

Application Notes
  1. The C398.4A antibody is useful for flow cytometric analysis and is able to costimulate T cell activation and proliferation. Additional reported applications (for the relevant formats) include: immunoprecipitation1, in vitro costimulation of T cell activation1,3,4, and spatial biology (IBEX)5,6. The LEAF™ purified antibody (Endotoxin < 0.1 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays (Cat. No. 313512).
Additional Product Notes

For the use of this antibody in spatial biology applications, we have partnered with Lunaphore Technologies for demonstration of our antibodies on the COMET™. The COMET™ platform is an automated, end-to-end spatial biology solution developed for rapid and flexible multiplex tissue profiling. More information on the COMET™ and a complete list of our antibodies that have been demonstrated on the COMET™ can be found here.

Application References

(PubMed link indicates BioLegend citation)
  1. Redoglia V, et al. 1996. Eur. J. Immunol. 26:2781. (FC IP Costim)
  2. Yagi J, et al. 2003. J. Immunol. 171:783. (FC)
  3. Arimura Y, et al. 2002. Int. Immunol. 14:555. (Costim)
  4. Arimura Y, et al. 2004. J. Biol. Chem. 279:11408. (Costim)
  5. Radtke AJ, et al. 2020. Proc Natl Acad Sci USA. 117:33455-33465. (SB) PubMed
  6. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
  1. Gubin MM, et al. 2018. Cell. 175:1014. PubMed
  2. Bengsch B et al. 2018. Immunity. 48(5):1029-1045 . PubMed
  3. Wagner J et al. 2019. Cell. 177(5):1330-1345 . PubMed
  4. Del Alcazar D, et al. 2019. Cell Rep. 28:3047. PubMed
  5. Overacre-Delgoffe AE, et al. 2017. Cell. 169:1130. PubMed
  6. Sprouse ML, et al. 2018. JCI Insight. 3:e97322. PubMed
  7. Roussey JA, et al. 2017. J Immunol. 199:3535. PubMed
  8. Kim SJ, et al. 2017. Nat Immunol. 18:1016. PubMed
  9. Hu J, et al. 2019. Am J Transl Res. 3.043055556. PubMed
  10. Tripathi T, et al. 2019. Immunohorizons. 0.201388889. PubMed
  11. Chng MHY, et al. 2020. Immunity. 51(6):1119-1135.e5.. PubMed
  12. Brenchley J, et al. 2012. Blood. 120:4172. PubMed
  13. Kinosada H, et al. 2017. PLoS Pathog. 10.1371/journal.ppat.1006120. PubMed
  14. Galbraith MD, et al. 2021. eLife. 10:00. PubMed
  15. Friebel E, et al. 2020. Cell. 181(7):1626-1642.e20. PubMed
  16. Schwabenland M, et al. 2021. Immunity. . PubMed
  17. Sullivan KD, et al. 2021. Cell Reports. 36(7):109527. PubMed
  18. Mann ER, et al. 2020. Sci Immunol. :5. PubMed
  19. Rouers A, et al. 2021. Cell Rep Med. 2:100278. PubMed
  20. Kennedy-Darling J, et al. 2021. Eur J Immunol. 51:1262. PubMed
  21. Levin SD, et al. 2020. Front Immunol. 10:3086. PubMed
  22. Sun Y, et al. 2020. J Immunol. 205:3358. PubMed
  23. Stensland ZC, et al. 2022. iScience. 25:103626. PubMed
  24. Prabakaran T, et al. 2021. EBioMedicine. 66:103314. PubMed
  25. Kaw S, et al. 2020. EMBO J. 39:e105594. PubMed
  26. Macmillan ML, et al. 2021. Blood Adv. 5:1425. PubMed
  27. Son YM, et al. 2020. Eur J Immunol. 50:1067. PubMed
  28. Liu CJ, et al. 2020. Neuro Oncol. 22:1276. PubMed
  29. Spoerl S, et al. 2021. Eur J Immunol. 51:1436. PubMed
  30. McIlwain DR, et al. 2021. Cell Host Microbe. 29:1828. PubMed
  31. McCarthy EE, et al. 2022. Cell Rep. 39:110815. PubMed
  32. Poon MML, et al. 2023. Nat Immunol. 24:309. PubMed
  33. Hailemichael Y, et al. 2022. Cancer Cell. 40:509. PubMed
  34. Liu T, et al. 2023. Cardiovasc Res. 119:1046. PubMed
  35. Diray-Arce J, et al. 2023. Cell Rep Med. 4:101079. PubMed
RRID
AB_416326 (BioLegend Cat. No. 313502)

Antigen Details

Structure
CD28/CTLA-4, 47-57 kD
Distribution

Activated T cells, subset of thymocytes

Function
Costimulates T cell activation, proliferation, humoral immune response
Ligand/Receptor
B7h/B7RP-1/GL-50
Cell Type
T cells, Thymocytes, Tregs
Biology Area
Costimulatory Molecules, Immunology
Molecular Family
CD Molecules
Antigen References

1. Redoglia V, et al. 1996. Eur. J. Immunol. 26:2781.
2. Hutloff A, et al. 1999. Nature 397:263.
3. Buonfiglio D, et al. 2000. Eur. J. Immunol. 30:3463.
4. Coyle AJ, et al. 2000. Immunity 13:95.

Gene ID
100048841 View all products for this Gene ID 29851 View all products for this Gene ID 64545 View all products for this Gene ID
UniProt
View information about CD278 on UniProt.org

Related FAQs

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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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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