PE anti-mouse/rat/human CD27 Antibody

Product Details

Verified Reactivity

Mouse, Rat, Human

Antibody Type

Monoclonal

Host Species

Armenian Hamster

Immunogen

Armenian hamster fibroblast line ARHO12 transfected with mouse CD27 cDNA

Formulation

Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide

Preparation

The antibody was purified by affinity chromatography, and conjugated with PE under optimal conditions.

Concentration

0.2 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 - 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 ≤ 0.25 µg per million cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.

Excitation Laser

Blue Laser (488 nm)
Green Laser (532 nm)/Yellow-Green Laser (561 nm)

Application Notes

Additional reported applications (for the relevant formats) include: functional assay^1,3 and immunohistomecial staining of acetone-fixed frozen sections². The Ultra-LEAF™ purified antibody (Endotoxin < 0.01 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays (Cat. Nos. 124243 & 124244).

Additional Product Notes

For the use of this antibody in spatial biology (SB), we have partnered with Bruker Spatial Biology Biosciences for demonstration of this antibody on their next-generation ChipCytometry instrument called the CellScape™. The CellScape platform is an end-to-end solution for highly multiplexed spatial omics. Combining an advanced, purpose-built imaging system with easy-to-use fluidics for walk-away automation, the CellScape system will accelerate your exploration into the rapidly evolving field of spatial biology. More information on the the Bruker Spatial Biology CellScape and a complete list of our antibodies that have been demonstrated on the instrument can be found here.

Application References

(PubMed link indicates BioLegend citation)

1. Gravestein LA, et al. 1995. Int. Immunol. 7:551. (FA)
2. Gravestein LA, et al. 1996. J. Exp. Med. 184:675. (IHC)
3. Takeda K, et al. 2000. J. Immunol. 164:1741. (FA)
4. Welner RS, et al. 2009. J. Immunol. 183:7768. PubMed
5. Vicetti Miguel RC, et al. 2012. J. Immunol. 189:3449. PubMed
6. White CA, et al. 2014. J Immunol. 193:5933. PubMed
7. Iwata S, et al. 2015. Lupus. 24:695. PubMed

Product Citations

1. Dong MB, et al. 2020. Cell. 178(5):1189-1204.e23.. PubMed
2. Liu L, et al. 2022. Nat Commun. 13:6740. PubMed
3. Wu R, et al. 2022. Nat Immunol. 23:1536. PubMed
4. Fu B et al. 2017. Immunity. 47(6):1100-1113 . PubMed
5. Liu X, et al. 2021. eLife. 0.416666666666667. PubMed
6. Beatson RE, et al. 2021. Cell Rep Med. 2:100473. PubMed
7. Xiong Y, et al. 2018. Int J Clin Exp Pathol. 3.803472222. PubMed
8. Cong J et al. 2018. Cell metabolism. 28(2):243-255 . PubMed
9. Tacconi C, et al. 2021. Cell Reports. 35(2):108993. PubMed
10. Shibata K, et al. 2011. Blood. 118:586. PubMed
11. Yu X, et al. 2019. Nat Commun. 10:574. PubMed
12. Ni J, et al. 2020. Immunity. 52(6):1075-1087.e8. PubMed
13. NULL, et al. 2022. Cell. 185:916. PubMed
14. Xiong Y, et al. 2019. BMC Immunol. 20:24. PubMed
15. Wagner AK, et al. 2022. iScience. 25:105137. PubMed
16. Wang J, et al. 2020. Cell. 183(7):1867-1883.e26. PubMed
17. Kujur W, et al. 2020. PLoS Pathog. 16:e1009132. PubMed

RRID

AB_1236459 (BioLegend Cat. No. 124209)
AB_1236459 (BioLegend Cat. No. 124210)

Antigen Details

Structure

TNFR superfamily, 45 kD

Distribution

Peripheral blood T cells, memory B cells, NK cells, thymocyte subset

Function

Costimulatory signal for T and B cell activation, T cell development

Interaction

TRAF2, TRAF5, CD27-binding protein (Siva)

Ligand/Receptor

CD70

Cell Type

B cells, NK cells, T cells, Thymocytes

Biology Area

Costimulatory Molecules, Immunology

Molecular Family

CD Molecules

Antigen References

1. Hintzen RQ, et al. 1994. Immunol. Today 15:307.
2. Akiba H, et al. 1998. J. Biol. Chem. 273:13353.
3. Kobata T, et al. 1995. P. Natl. Acad. Sci. USA 92:11249.
4. Hendriks J, et al. 2000. Nat. Immunol. 1:433
5. Hintzen RQ, et al. 1994. Int. Immunol. 6:477
6. Camerini D, et al. 1991. J. Immunol. 147:3165

Gene ID

21940 View all products for this Gene ID 939 View all products for this Gene ID 500318 View all products for this Gene ID

UniProt

View information about CD27 on UniProt.org

Documentation

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

Reviews

Related Protocols

• Cell Surface Flow Cytometry Staining Protocol

Related Pages & Pathways

Related FAQs

What type of PE do you use in your conjugates?

We use R-PE in our conjugates.

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