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Alexa Fluor® 488 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
1_C398_4A_Alx488_072007
PHA-stimulated human peripheral blood lymphocytes (3 days) stained with C398.4A Alexa Fluor® 488
  • 1_C398_4A_Alx488_072007
    PHA-stimulated human peripheral blood lymphocytes (3 days) stained with C398.4A Alexa Fluor® 488
  • 6_Human_LN_ICOS_CD35
    Confocal image of human lymph node sample acquired using the IBEX method of highly multiplexed antibody-based imaging: ICOS (yellow) in Cycle 6, CD35 (purple) in Cycle 7. 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).
Compare all formats See Alexa Fluor® 488 spectral data
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313514 100 µg 229 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

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 and conjugated with Alexa Fluor® 488 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
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 ≤ 0.5 µg per 106 cells in 100 µl volume. It is recommended that reagents be titrated for optimal performance in the particular application.

* Alexa Fluor® 488 has a maximum emission of 519 nm when it is excited at 488 nm.


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

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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

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

(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. Matsui K, et al. 2015. PLoS One. 10: 0137195. PubMed
  2. Yue X, et al. 2019. Nat Commun. 10:2011. PubMed
  3. Shuwa HA, et al. 2021. Med. 2(6):720-735.e4. PubMed
  4. Masuda N, et al. 2021. BMC Cancer. 21:1269. PubMed
  5. Lederer K, et al. 2020. Immunity. 53(6):1281-1295.e5. PubMed
RRID
AB_2122584 (BioLegend Cat. No. 313514)

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.

Other Formats

View All CD278 Reagents Request Custom Conjugation
Description Clone Applications
Purified anti-human/mouse/rat CD278 (ICOS) C398.4A FC,IP,Costim,SB
Biotin anti-human/mouse/rat CD278 (ICOS) C398.4A FC
FITC anti-human/mouse/rat CD278 (ICOS) C398.4A FC
PE anti-human/mouse/rat CD278 (ICOS) C398.4A FC
APC anti-human/mouse/rat CD278 (ICOS) C398.4A FC
Alexa Fluor® 488 anti-human/mouse/rat CD278 (ICOS) C398.4A FC,SB
Alexa Fluor® 647 anti-human/mouse/rat CD278 (ICOS) C398.4A FC,IHC-F
PerCP/Cyanine5.5 anti-human/mouse/rat CD278 (ICOS) C398.4A FC
PE/Cyanine7 anti-human/mouse/rat CD278 (ICOS) C398.4A FC
Pacific Blue™ anti-human/mouse/rat CD278 (ICOS) C398.4A FC
Brilliant Violet 421™ anti-human/mouse/rat CD278 (ICOS) C398.4A FC
Brilliant Violet 510™ anti-human/mouse/rat CD278 (ICOS) C398.4A FC
APC/Cyanine7 anti-human/mouse/rat CD278 (ICOS) C398.4A FC
PE/Dazzle™ 594 anti-human/mouse/rat CD278 (ICOS) C398.4A FC
Alexa Fluor® 700 anti-human/mouse/rat CD278 (ICOS) C398.4A FC
APC/Fire™ 750 anti-human/mouse/rat CD278 (ICOS) C398.4A FC
Brilliant Violet 785™ anti-human/mouse/rat CD278 (ICOS) C398.4A FC
Brilliant Violet 605™ anti-human/mouse/rat CD278 (ICOS) C398.4A FC
Ultra-LEAF™ Purified anti-human/mouse/rat CD278 (ICOS) C398.4A FC,IP,Costim
GoInVivo™ Purified anti-human/mouse/rat CD278 (ICOS) C398.4A FC
Brilliant Violet 711™ anti-human/mouse/rat CD278 (ICOS) C398.4A FC
Brilliant Violet 650™ anti-human/mouse/rat CD278 (ICOS) C398.4A FC
TotalSeq™-B0171 anti-human/mouse/rat CD278 (ICOS) C398.4A PG
TotalSeq™-C0171 anti-human/mouse/rat CD278 (ICOS) C398.4A PG
TotalSeq™-A0171 anti-human/mouse/rat CD278 (ICOS) C398.4A PG
Brilliant Violet 750™ anti-human/mouse/rat CD278 (ICOS) C398.4A FC
TotalSeq™-D0171 anti-human/mouse/rat CD278 (ICOS) Antibody C398.4A PG
PE/Cyanine5 anti-human/mouse/rat CD278 (ICOS) C398.4A FC
Spark Red™ 718 anti-human/mouse/rat CD278 (ICOS) C398.4A FC
Go To Top Version: 5    Revision Date: 04.19.2022

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

 

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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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