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Purified anti-human CD66b Antibody

Pricing & Availability
Clone
6/40c (See other available formats)
Regulatory Status
RUO
Other Names
CEACAM8, CD67, CGM6, NCA-95
Isotype
Mouse IgG1, κ
Ave. Rating
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Product Citations
publications
A_6slash40c_pure_CD66b_Antibody_012918
Human peripheral blood granulocytes was stained with CD66b (clone 6/40c) purified (filled histogram) or mouse IgG1, κ isotype control (clone MOPC-21) purified (open histogram) followed by Poly4053 PE.
  • A_6slash40c_pure_CD66b_Antibody_012918
    Human peripheral blood granulocytes was stained with CD66b (clone 6/40c) purified (filled histogram) or mouse IgG1, κ isotype control (clone MOPC-21) purified (open histogram) followed by Poly4053 PE.
  • B_6slash40c_PURE_CD66b_Antibody_IHC-P_032018
    Human paraffin-embedded spleen tissue slices were prepared with a standard protocol of deparaffinization and rehydration. Antigen retrieval was done with Sodium Citrate H.I.E.R., 1x 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 purified anti-human CD66b (clone 6/40c) antibody overnight at 4°C. On the next day, tissue was incubate with Alexa Fluor® 594 Goat anti-mouse IgG (clone poly4053) antibody (red). Nuclei were counter-stained with DAPI (blue). The image was scanned with a 10X objective and stitched with MetaMorph® software.
  • C_6slash40c_PURE_CD66b_Antibody_SB_111623
    SeqIF™ (sequential immunofluorescence) staining on COMET™ of Purified anti-CD66b (clone 6/40c, yellow) on formalin-fixed paraffin-embedded human tonsil tissue at 5 µg/mL. Alexa Fluor™ Plus 647 Goat anti-Mouse IgG antibody (Lunaphore, Cat. No. DR647MS) was used as a secondary antibody. Nuclei were counterstained with DAPI (blue). Tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing.
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392902 100 µg 141 CHF
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Description

CD66b is a 95-100 kD glycosylphosphatidylinositol (GPI)-linked protein also known as CD67, CGM6, and NCA-95. CD66b is a member of the immunoglobulin superfamily, carcinoembryonic antigen (CEA)-like subfamily. CD66b, expressed on granulocytes, has been reported to induce activation in neutrophils and to be involved in heterophilic adhesion with CD66c.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Recombinant soluble human CEACAM8-Fc produced in HEK293 cells
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/ml
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
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 ≤ 0.5 µg per million cells in 100 µl volume. For immunohistochemistry, 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.

Additional Product Notes

For the use of this antibody in spatial biology applications, we have partnered with Lunaphore Technologies for demonstration of our antibodies on the COMET™. The COMET™ platform is an automated, end-to-end spatial biology solution developed for rapid and flexible multiplex tissue profiling. More information on the COMET™ and a complete list of our antibodies that have been demonstrated on the COMET™ can be found here.

Application References

(PubMed link indicates BioLegend citation)
  1. Singer, et al. 2014. PLoS One. 9(4):e94106. PMID: 24743304
  2. Klaile, et al. 2013. Respir Res. 14:85. PMID 23941132
Product Citations
  1. Fox-Fisher I, et al. 2022. Med (N Y). 3:468. PubMed
  2. Vondra S, et al. 2023. Cell Rep. 42:111977. PubMed
  3. He M, et al. 2021. J Gynecol Oncol. e32:32. PubMed
  4. Riquelme E et al. 2019. Cell. 178(4):795-806 . PubMed
  5. Zheng Y, et al. 2022. iScience. 25:103785. PubMed
RRID
AB_2728422 (BioLegend Cat. No. 392902)

Antigen Details

Structure
Ig superfamily, CEA antigen group, GPI-linked glycoprotein
Distribution

Granulocytes

Function
Cell adhesion, neutrophil activation
Ligand/Receptor
CD66c
Cell Type
Granulocytes
Biology Area
Immunology
Molecular Family
Adhesion Molecules, CD Molecules
Antigen References
  1. Kuijpers T, et al. 1993. J Immunol. 151:4934.
  2. Kuroki M, et al. 1992. J Leuk Biol. 52:551.
Gene ID
1088 View all products for this Gene ID
UniProt
View information about CD66b 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: 2    Revision Date: 11.16.2023

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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