Alexa Fluor® 488 anti-human IgD Antibody

Pricing & Availability
Clone
IA6-2 (See other available formats)
Regulatory Status
RUO
Other Names
Ig delta chain C region
Isotype
Mouse IgG2a, κ
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Product Citations
publications
1_IA6-2_AF488_1_022712
Human peripheral blood lymphocytes were stained with CD19 Brilliant Violet 421™ and IgD (clone IA6-2) Alexa Fluor® 488 (top) or mouse IgG2a, κ Alexa Fluor® 488 isotype control (bottom).
  • 1_IA6-2_AF488_1_022712
    Human peripheral blood lymphocytes were stained with CD19 Brilliant Violet 421™ and IgD (clone IA6-2) Alexa Fluor® 488 (top) or mouse IgG2a, κ Alexa Fluor® 488 isotype control (bottom).
  • 2_IA6-2_AF488_2_022712
  • 3_Human_LN_IgD_PD-1
    Confocal image of human lymph node sample acquired using the IBEX method of highly multiplexed antibody-based imaging: IgD (blue) in Cycle 2, PD-1 (green) in Cycle 5. 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).
  • 4_60_Mouse_Lymph_Node_PD1_CD23_IgD
    Mice were injected subcutaneously with sheep red blood cells in a volume of 25 µL per site on days 0 and 4 and harvested on day 11. Confocal image of C57BL/6 mouse lymph node acquired using the IBEX method of highly multiplexed antibody-based imaging: PD-1 (green) in Cycle 1, CD23 (magenta) in Cycle 7, and IgD (blue) in Cycle 10. 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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348215 25 tests £76
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348216 100 tests £170
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Description

IgD, a member of the immunoglobulin (Ig) family, is expressed in naïve B cells. It has 3 Ig-like domains and exists in a transmembrane and a soluble form. In general, IgD is not secreted and usually its expression is lost after the Ig isotype switch. After antigen binding, IgD signals through the CD79a/CD79b (Igα/Igβ) heterodimer, resulting in the activation of the B cell.

Product Details
Technical Data Sheet (pdf)

Product Details

Reactivity
Human
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Human IgD
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® 488 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.

* Alexa Fluor® 488 has a maximum emission of 519 nm when it is excited at 488 nm.


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

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Excitation Laser
Blue Laser (488 nm)
Application Notes

Additional reported applications (for the relevant formats) include: immunohistochemical staining of paraformaldehyde fixed frozen sections4 and spatial biology (IBEX)5,6.

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

(PubMed link indicates BioLegend citation)
  1. Chen K, et al. 2009. Nat. Immunol. 10:889.
  2. Lee CH, et al. 2005. J. Exp. Med. 203:63.
  3. Sutter JA, et al. 2008. Clin. Immunol. 126:282.
  4. Li H and Pauza CD. 2015. Eur. J. Immunol. 45:298. (IHC)
  5. Radtke AJ, et al. 2020. Proc Natl Acad Sci USA. 117:33455-33465. (SB) PubMed
  6. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
  1. Japp AS, et al. 2021. Cell. 184(3):827-839.e14. PubMed
  2. Harrington WE, et al. 2021. Cell Reports Medicine. 2(4):100253. PubMed
  3. Setliff I, et al. 2018. Cell Host Microbe. 23:845. PubMed
  4. Sajadi MM, et al. 2018. Cell. 173:1783. PubMed
  5. Powell WE, et al. 2018. Diabetologia. 61:1794. PubMed
RRID
AB_11150397 (BioLegend Cat. No. 348215)
AB_11150595 (BioLegend Cat. No. 348216)

Antigen Details

Structure
Exists in a transmembranal and a soluble form
Distribution

Naïve B cells

Function
Antigen binding, B cell activation
Interaction
The CD79a/CD79b heterodimer
Cell Type
B cells
Biology Area
Immunology
Antigen References

1. Geisberger R, et al. 2006. Immunology 118:429.
2. Weller S, et al. 2005. Eur. J. Immunol. 35:2789.
3. Brandtzaeg P and Johansen FE. 2005. Immunol. Rev. 206:32.

Gene ID
3495 View all products for this Gene ID
UniProt
View information about IgD 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.

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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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