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Alexa Fluor® 488 anti-mouse IgD Antibody

Pricing & Availability
Clone
11-26c.2a (See other available formats)
Regulatory Status
RUO
Other Names
Immunoglobulin D
Isotype
Rat IgG2a, κ
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Product Citations
publications
11-26c.2a_AF488_1_021111
C57BL/6 splenocytes were stained with B220 APC and IgD (11-26c.2a) Alexa Fluor® 488 (top) or rat IgG2a, κ Alexa Fluor® 488 isotype control (bottom).
  • 11-26c.2a_AF488_1_021111
    C57BL/6 splenocytes were stained with B220 APC and IgD (11-26c.2a) Alexa Fluor® 488 (top) or rat IgG2a, κ Alexa Fluor® 488 isotype control (bottom).
  • 11-26c.2a_AF488_2_021111
  • 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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405717 25 µg 90€
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405718 100 µg 212€
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Description

Surface IgD is an important B cell differentiation marker.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Mouse
Antibody Type
Monoclonal
Host Species
Rat
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 488 under optimal conditions.
Concentration
0.5 mg/ml
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 ≤0.5 µg per million cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.

* 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

The 11-26c.2a antibody reacts with immunoglobulin D in all tested mouse haplotypes. The antibody binds membrane IgD expressed on most B cells. The 11-26c.2a antibody neither induces proliferation of splenic B cells nor induces B cell activation. Additional reported applications (for the relevant formats) include: immunohistochemical staining of acetone-fixed frozen sections2,3, and spatial biology (IBEX)10,11.

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. Nitschke L, et al. 1993. P. Natl. Acad. Sci. USA 90:1887. (FC)
  2. Weih D, et al. 2001. J. Immunol. 167:1909. (IHC)
  3. Koni PA, et al. 2001. J. Exp. Med. 193:741. (IHC)
  4. Ahuja A, et al. 2007. J. Immunol. 179:3351. (FC) PubMed
  5. Haynes NM, et al. 2007. J. Immunol. 179:5099. (FC)
  6. Good-Jacobson KL, et al. 2010. Nat. Immunol. 11:535. (FC) PubMed
  7. Tomayko MM, et al. 2010. J. Immunol. 185:7146. PubMed
  8. Park SY, et al. 2013. J. Immunol. 190:1094. PubMed
  9. Rouaud P, et al. 2014. J Exp Med. 211:975. PubMed
  10. Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed
  11. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
  1. Shenoy AT, et al. 2022. Cytometry A. 101:892. PubMed
  2. Voss LF, et al. 2023. Front Immunol. 13:1021370. PubMed
  3. Rouaud P, et al. 2014. J Exp Med. 211:975. PubMed
  4. Kim CJ, et al. 2018. Immunity. 49:1034. PubMed
  5. Sarapulov AV, et al. 2020. Front Immunol. 11:599. PubMed
  6. Ly A, et al. 2020. Cell Reports. 29(8):2257-2269.e6.. PubMed
  7. Glaros V, et al. 2021. Immunity. 54:2005. PubMed
  8. Béguelin W, et al. 2020. Cancer Cell. 37(5):655-673. PubMed
  9. Haase P, et al. 2020. Eur J Immunol. 50:558. PubMed
  10. Tuong ZK, et al. 2021. Cell Rep. 37:110132. PubMed
  11. Katsuyama T, et al. 2019. J Clin Invest. 129:5411. PubMed
RRID
AB_10730618 (BioLegend Cat. No. 405717)
AB_10730618 (BioLegend Cat. No. 405718)

Antigen Details

Structure
Ig family
Distribution

B cells

Function
B cell differentiation
Cell Type
B cells
Biology Area
Immunology
Gene ID
380797 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.

Go To Top Version: 3    Revision Date: 04.18.2022

For Research Use Only. Not for diagnostic or therapeutic 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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