PE anti-human CD57 Antibody

Product Details

Verified Reactivity

Human

Antibody Type

Monoclonal

Host Species

Mouse

Immunogen

Membrane extract of human lymphoblastoid cell line HSB-2.

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 PE 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 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 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)
Green Laser (532 nm)/Yellow-Green Laser (561 nm)

Application Notes

Additional reported applications for the relevant formats include: Western blotting¹.

Additional Product Notes

This product has been verified for IHC-P (Immunohistochemistry - formalin-fixed paraffin-embedded tissues) on the NanoString GeoMx® Digital Spatial Profiler. The GeoMx® enables researchers to perform spatial analysis of protein and RNA targets in FFPE and fresh frozen human and mouse samples. For more information about our spatial biology products and the GeoMx® platform, please visit our spatial biology page.

Application References

(PubMed link indicates BioLegend citation)

1. Yoshihara Y, et al. 1991. J. Cell Biol. 115:731. (WB)
2. Abo T, et al. 1981. J. Immunol. 127:1024.
3. Abo T, et al. 1982. J. Immunol. 129:1752.
4. Abo T, et al. 1982. J. Immunol. 129:1758.

Product Citations

1. Jarosch S, et al. 2022. STAR Protoc. 3:101374. PubMed
2. Scherpenisse M, et al. 2021. MBio. 12:. PubMed
3. Salumets A, et al. 2022. Aging Cell. 21:e13607. PubMed
4. Jia L, et al. 2022. Ann Transl Med. 10:482. PubMed

RRID

AB_2562758 (BioLegend Cat. No. 359611)
AB_2562759 (BioLegend Cat. No. 359612)

Antigen Details

Structure

Oligosaccharide antigenic determinant present on a variety of proteins, lipids, and chrondroitin sulfate proteoglycans, 100-115 kD. The antigen is conserved across species.

Distribution

CD57 antigen is expressed on a subset of peripheral blood lymphocytes including a subset of NK cells and a subset of CD8⁺ T cells. Also expressed on neural cells and striated muscle.

Function

Unknown function, may be involved in cell-matrix interactions.

Ligand/Receptor

Binds to L-selectin and P-selectin in a calcium-dependent manner, also binds to second globular domain of E8 laminin fragment.

Cell Type

Lymphocytes, NK cells, T cells

Biology Area

Costimulatory Molecules, Immunology

Molecular Family

CD Molecules

Antigen References

1. Schubert J, et al. 1989. In Leucocyte Typing IV (Knapp W, ed) Oxford University Press Oxford pp 711-714.
2. Palmer BE, et al. 2005. J. Immunol. 175:8415.
3. Schachner M, et al. 1995. Trends Neurosci. 18:183.
4. Wood KL, et al. 2005. Clin. Immunol. 117:294.

Gene ID

27087 View all products for this Gene ID

UniProt

View information about CD57 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