TotalSeq™-Bn1320 anti-human CD8a Antibody

Pricing & Availability
Clone
C8/144B (See other available formats)
Regulatory Status
RUO
Other Names
T8, Leu2
Isotype
Mouse IgG1, κ
Barcode Sequence
GTGTACGCCTTGTAC
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Product Citations
publications
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372911 10 µg $369
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Description

CD8a is a 32-34 kD type I glycoprotein. It forms a homodimer (CD8a/a) or heterodimer (CD8a/b) with CD8b. CD8, also known as T8 and Leu2, is a member of the immunoglobulin superfamily found on the majority of thymocytes, a subset of peripheral blood T cells, and NK cells (which express almost exclusively CD8a homodimers). CD8 acts as a co-receptor with MHC class I-restricted T cell receptors in antigen recognition and T cell activation, and has been shown to play a role in thymic differentiation. Two domains in CD8a are important for function: the extracellular IgSF domain binds the α3 domain of MHC class I and the cytoplasmic CXCP motif binds the tyrosine kinase p56 Lck.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Human, Mouse, Rat
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
A 13 amino acid synthetic peptide from the C-terminal cytoplasmic domain of the alpha chain of the human CD8 molecule.
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and EDTA
Preparation
The antibody was purified by chromatography and conjugated with TotalSeq™-Bn oligomer under optimal conditions.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C. Do not freeze.
Application

SB - Quality tested

Recommended Usage

Each lot of this antibody is quality control tested by immunofluorescent staining in formalin-fixed, paraffin-embedded (FFPE) lymphoid tissue, and the oligomer sequence is confirmed by sequencing. TotalSeq™-Bn antibodies are compatible with the 10x Visium CytAssist Gene and Protein Expression Assay.

To maximize performance, it is strongly recommended that the reagent be titrated for each application, and that you centrifuge the antibody dilution at 14,000xg at 2 − 8°C for 10 minutes before use. Carefully pipette out the liquid avoiding the bottom of the tube when handling. To determine and optimize dilutions for the addition of Totalseq™-Bn antibodies into pre-designed antibody panels, refer to 10x Genomics Custom Add-on Antibody Optimization guide.


Buyer is solely responsible for determining whether Buyer has all intellectual property rights that are necessary for Buyer's intended uses of the BioLegend TotalSeq™ products. For example, for any technology platform Buyer uses with TotalSeq™, it is Buyer's sole responsibility to determine whether it has all necessary third party intellectual property rights to use that platform and TotalSeq™ with that platform.
Application Notes

Additional reported applications (for the relevant formats) include activation3, flow cytometry4, immunohistochemical staining of frozen tissue sections2, and Western blotting5.

Additional Product Notes

TotalSeq™-Bn reagents are designed to profile protein levels following an optimized protocol in spatial transcriptomics. Compatible spatial biology devices (e.g. Imaging System, 10x Genomics Visium Spatial CytAssist Gene and Protein Expression instruments and reagents) and sequencer (e.g. Illumina analyzers) are required. TotalSeq™-B reagents are not compatible with the 10x Genomics Visium system. The complete barcode sequence may be provided upon request. Please contact technical support for more information, or visit TotalSeq™-Bn Reagents for 10x Genomics Visium CytAssist Gene and Protein Assay

Application References

(PubMed link indicates BioLegend citation)
  1. McQuitty E, et al. 2014. J. Cutan. Pathol. 41:88. (IHC-P)
  2. Petukhova L, et al. 2010. Nature 466:113. (IHC-F)
  3. Clement M, et al. 2011. J. Immunol. 187:654. (Activ)
  4. MarTchal R, et al. 2010. BMC Cancer 10:340. (FC)
  5. Pontiggia L, et al. 2008. J. Invest. Dermatol. 129:480. (WB)
RRID
AB_3083195 (BioLegend Cat. No. 372911)

Antigen Details

Structure
Ig superfamily, homodimer or heterodimer with CD8β.
Distribution

Majority of thymocytes, T cell subset, NK cells.

Function
MHC class I co-receptor, thymic differentiation, T-cell activation.
Ligand/Receptor
MHC class I molecules.
Cell Type
NK cells, T cells, Thymocytes
Biology Area
Immunology
Molecular Family
CD Molecules
Antigen References

1. McQuitty E, et al. 2014. J. Cutan. Pathol. 41:88.

Gene ID
100387674 View all products for this Gene ID
UniProt
View information about CD8a 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: 1    Revision Date: 08/24/2023

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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