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

Product Details

Verified Reactivity

Human, Mouse, Rat

Reported Reactivity

African Green, Baboon, Cynomolgus, Rhesus, Pig

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 activation^1,3,4, 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. Hu J, et al. 2019. Am J Transl Res. 3.043055556. PubMed
2. Rouers A, et al. 2021. Cell Rep Med. 2:100278. PubMed
3. Friebel E, et al. 2020. Cell. 181(7):1626-1642.e20. PubMed
4. Wagner J et al. 2019. Cell. 177(5):1330-1345 . PubMed
5. Sullivan KD, et al. 2021. Cell Reports. 36(7):109527. PubMed
6. Son YM, et al. 2020. Eur J Immunol. 50:1067. PubMed
7. Mann ER, et al. 2020. Sci Immunol. :5. PubMed
8. Overacre-Delgoffe AE, et al. 2017. Cell. 169:1130. PubMed
9. Kim SJ, et al. 2017. Nat Immunol. 18:1016. PubMed
10. Sprouse ML, et al. 2018. JCI Insight. 3:e97322. PubMed
11. Tripathi T, et al. 2019. Immunohorizons. 0.201388889. PubMed
12. Sun Y, et al. 2020. J Immunol. 205:3358. 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. Del Alcazar D, et al. 2019. Cell Rep. 28:3047. PubMed
16. Brenchley J, et al. 2012. Blood. 120:4172. PubMed
17. Gubin MM, et al. 2018. Cell. 175:1014. PubMed
18. Macmillan ML, et al. 2021. Blood Adv. 5:1425. PubMed
19. Chng MHY, et al. 2020. Immunity. 51(6):1119-1135.e5.. PubMed
20. Levin SD, et al. 2020. Front Immunol. 10:3086. PubMed
21. Bengsch B et al. 2018. Immunity. 48(5):1029-1045 . PubMed
22. Roussey JA, et al. 2017. J Immunol. 199:3535. PubMed
23. Schwabenland M, et al. 2021. Immunity. . PubMed
24. Kennedy-Darling J, et al. 2021. Eur J Immunol. 51:1262. PubMed
25. Stensland ZC, et al. 2022. iScience. 25:103626. PubMed
26. Kaw S, et al. 2020. EMBO J. 39:e105594. PubMed
27. Prabakaran T, et al. 2021. EBioMedicine. 66:103314. 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

Documentation

• Technical Data Sheet (pdf)
• Certificate of Analysis
• Safety Data Sheet

Reviews

Related Pages & Pathways

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.

Other Formats

Certificate of Analysis