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Alexa Fluor® 647 anti-human CD57 Antibody

Pricing & Availability
Clone
HNK-1 (See other available formats)
Regulatory Status
RUO
Other Names
HNK-1, NK-1, Leu-7
Isotype
Mouse IgM, κ
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Product Citations
publications
1_HNK-1_AF647_CD57_Antibody_FC_1_111313
  • 1_HNK-1_AF647_CD57_Antibody_FC_1_111313
  • 2_HNK-1_AF647_CD57_Antibody_FC_2_111313
  • 3_HNK-1_A647_CD57_Antibody_IHC_031921
    Human paraffin-embedded cerebellum tissue slices were prepared with a standard protocol of deparaffinization and rehydration. Antigen retrieval was done with Citrate Buffered 1X (10mM, pH 6.0) at 95°C for 40 minutes. Tissue was washed with PBS/0.05% Tween 20 twice for five minutes and blocked with 5% FBS and 0.2% gelatin for 30 minutes. Then, the tissue was stained with 10 µg/mL of Alexa Fluor® 488 anti-TUBB3 (clone AA10) antibody (green) and Alexa Fluor® 647 anti-CD57 (clone HNK-1) antibody (red) over night at 4°C. The nuclei were counter-stained with DAPI. The image was scanned with a 10X objective and stitched with software.
Compare all formats See Alexa Fluor® 647 spectral data
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359613 25 tests 100€
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359614 100 tests 212€
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Description

CD57, also known as HNK-1, NK-1, and Leu-7 is a 100-115 kD oligosaccharide antigenic determinant expressed on a variety of proteins, lipids, and chondroitin sulfate proteoglycans. CD57 is expressed on a subset of peripheral blood lymphocytes, including NK cells and CD8+ T cells, and is also expressed on neural cells and striated muscle. CD57 is not expressed on red blood cells, granulocytes, monocytes, or platelets. While the function of CD57 is unknown, binding to L-selectin, P-selectin, and a fragment of laminin suggests that CD57 may be involved in cell-matrix interactions. CD57 is increased in some disease states associated with CD4/CD8 imbalances (AIDS, autoimmune disease, viral infections, and allograft transplants).

Product Details
Technical Data Sheet (pdf)

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 Alexa Fluor® 647 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
IHC-P - Verified
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. For immunohistochemistry on formalin-fixed paraffin-embedded tissue sections, a concentration range of  5.0 - 10 µg/mL is suggested. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 647 has a maximum emission of 668 nm when it is excited at 633 nm / 635 nm.


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

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Excitation Laser
Red Laser (633 nm)
Application Notes

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

Additional Product Notes

This product has been verified for IHC-F (Immunohistochemistry - frozen tissue sections) and 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
  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.
RRID
AB_2562857 (BioLegend Cat. No. 359613)
AB_2562857 (BioLegend Cat. No. 359614)

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

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.

Go To Top Version: 4    Revision Date: 01/24/2024

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

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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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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