PE anti-human CD35 Antibody

Pricing & Availability
Clone
E11 (See other available formats)
Regulatory Status
RUO
Workshop
III 204
Other Names
C3b/C4b receptor, complement receptor type 1 (CR1)
Isotype
Mouse IgG1, κ
Ave. Rating
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Product Citations
publications
1_E11_PE_040810.jpg
Human peripheral blood lymphocytes stained with E11 PE
  • 1_E11_PE_040810.jpg
    Human peripheral blood lymphocytes stained with E11 PE
  • 6_Human_LN_ICOS_CD35
    Confocal image of human lymph node sample acquired using the IBEX method of highly multiplexed antibody-based imaging: ICOS (yellow) in Cycle 6, CD35 (purple) in Cycle 7. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
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333405 25 tests 88€
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333406 100 tests 194€
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Description

CD35 is a type I single chain of glycoprotein, also known as C3b/C4b receptor, Complement Receptor type 1 or CR1. Four molecular weight allotypes (160kD, 190kD, 220kD, and 250kD) have been described. CD35 is expressed on granulocytes, monocytes, B cells, erythrocytes, and follicular dendritic cells, as well as subsets of NK and T cells. CD35 binds complement C3b, C4b, or iC3, and iC4, and plays important roles in both innate and adoptive immune response via mediating phagocytosis by granulocytes and monocytes. CD35 has also been reported to inhibit T-cell proliferation.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human
Reported Reactivity
African Green, Baboon, Cynomolgus, Rhesus
Antibody Type
Monoclonal
Host Species
Mouse
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, please enter the lot number in our Concentration and Expiration Lookup or Certificate of Analysis online tools.)
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 - Reported in the literature, not verified in house

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: spatial biology (IBEX)4,5.

Additional Product Notes

Iterative Bleaching Extended multi-pleXity (IBEX) is a fluorescent imaging technique capable of highly-multiplexed spatial analysis. The method relies on cyclical bleaching of panels of fluorescent antibodies in order to image and analyze many markers over multiple cycles of staining, imaging, and, bleaching. It is a community-developed open-access method developed by the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).

Application References
  1. Hogg N, et al. 1984. Eur. J. Immunol. 14:236
  2. Zabeleta J, et al. 1998. Clin Diag. Lab. Immunol. 5:690
  3. Pascual M, et. al. 1994. Eur. J. Immunol. 24:702
  4. Radtke AJ, et al. 2020. Proc Natl Acad Sci USA. 117:33455-33465. (SB) PubMed
  5. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
  1. Flores E, et al. 2017. Infect Immun . 10.1128/IAI.01043-16. PubMed
  2. Bakke S,et al. 2017. J Immunol. 10.4049/jimmunol.1700302. PubMed
  3. Crane A, et al. 2018. Alzheimers Dement. 14:243. PubMed
  4. Magri G et al. 2017. Immunity. 47(1):118-134 . PubMed
  5. Stojkov D, et al. 2017. J Cell Biol. 216:4073. PubMed
RRID
AB_2085021 (BioLegend Cat. No. 333405)
AB_2292231 (BioLegend Cat. No. 333406)

Antigen Details

Structure
Type I glycoprotein with four allotypes, 160kD, 190kD, 220kD, 250kD
Distribution

Granulocytes, monocytes, B cells, erythrocytes, follicular dendritic cells, subsets of NK and T cells

Function
Adhesion, mediate phagocytosis, inhibit T-cell proliferation
Ligand/Receptor
C3b, iC3, C4b, iC4
Cell Type
B cells, Dendritic cells, Erythrocytes, Granulocytes, Monocytes, NK cells, T cells
Biology Area
Cell Biology, Complement, Immunology, Neuroinflammation, Neuroscience
Molecular Family
CD Molecules
Antigen References

1. Zola Heddy, et al. Eds. 2007. Leukocyte and Stromal Cell Molecules:The CD markers. WILEY-LISS
2. Klickstein LB, et al. 1988. J. Exp. Med. 168:1699

Gene ID
1378 View all products for this Gene ID
UniProt
View information about CD35 on UniProt.org

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.

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*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/ordering#license). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products, reverse engineer functionally similar materials, or to provide a service to third parties without written approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.

 

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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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