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PE anti-IRF4 Antibody

Pricing & Availability
Clone
IRF4.3E4 (See other available formats)
Regulatory Status
RUO
Other Names
Interferon regulatory factor 4
Isotype
Rat IgG1, κ
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Product Citations
publications
1_IRF4.3E4_PE_IRF4_Antibody_FC_011614.jpg
PHA-stimulated (3 days) or freshly isolated human peripheral blood lymphocytes were stained with CD4 APC, this fixed and permeabilized with True-Nuclear™ Transcription Factor Buffer Set (Cat. No. 424401). Cells were then stained with IRF4 (clone IRF4.3E4) PE (open histogram with solid line for fresh cells, filled histogram for stimulated cells) or rat IgG1, κ PE isotype control (dash line for fresh cell, dot line for stimulated cells). Data shown was gated on CD4+ lymphocyte population.
  • 1_IRF4.3E4_PE_IRF4_Antibody_FC_011614.jpg
    PHA-stimulated (3 days) or freshly isolated human peripheral blood lymphocytes were stained with CD4 APC, this fixed and permeabilized with True-Nuclear™ Transcription Factor Buffer Set (Cat. No. 424401). Cells were then stained with IRF4 (clone IRF4.3E4) PE (open histogram with solid line for fresh cells, filled histogram for stimulated cells) or rat IgG1, κ PE isotype control (dash line for fresh cell, dot line for stimulated cells). Data shown was gated on CD4+ lymphocyte population.
  • 2_7_Human_LN_CD31_CD138_IRF4
    Confocal image of human lymph node sample acquired using the IBEX method of highly multiplexed antibody-based imaging: CD31 (magenta) in Cycle 2, CD138 (blue) in Cycle 2, and IRF4 (green) in Cycle 6. 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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646403 25 µg $129
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646404 100 µg $335
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Description

The IRF family consists of at least nine members. IRF4 and IRF8 are highly homologous to each other and also redundant in function. IRF4 is critical for Th2 and Th17 development. Together with TRF8, it plays an essential role in macrophage and dendritic cell development and function. IRF4 is also reported to be essential for pre-B cell development, receptor editing, germinal center reactor and plasma cell differentiation.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Mouse, Human
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
GST fusion protein containing C-terminal of murine IRF4
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography and conjugated with PE under optimal conditions.
Concentration
0.2 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

ICFC - Quality tested
SB - Reported in the literature, not verified in house
Recommended Usage

Each lot of this antibody is quality control tested by intracellular immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is ≤0.25 µg per million cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.

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)3,4.

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. Zheng Y, et al. 2009. Nature 458:351
  2. Yin SY, et al. 2011. Exp Cell Res. 317:2210. PubMed
  3. Radtke AJ, et al. 2020. Proc Natl Acad Sci USA. 117:33455-33465. (SB) PubMed
  4. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
  1. Rivas MA, et al. 2021. Nat Immunol. 22:240. PubMed
  2. Hu Q, et al. 2022. Cell Rep. 40:111035. PubMed
  3. Arnold J, et al. 2015. Cell Death Differ. 23: 853-864. PubMed
  4. Webb LMC, et al. 2021. Aging Cell. 20:e13295. PubMed
  5. Lee JL, et al. 2022. Aging Cell. 21:e13692. PubMed
  6. Utley A, et al. 2020. Cell Rep. 31:107815. PubMed
  7. Holly Bachus et al. 2018. Immunity. 50(1):225-240 . PubMed
  8. Luo W et al. 2018. Immunity. 48(2):313-326 . PubMed
  9. Leylek R, et al. 2019. Cell Rep. 29:3736. PubMed
  10. Kurniawan H, et al. 2020. Cell Metabolism. 31(5):920-936. PubMed
  11. Trefzer A, et al. 2021. Cell Reports. 34(6):108748. PubMed
  12. Shih T, et al. 2019. Front Immunol. 10:1652. PubMed
  13. Yang BH, et al. 2020. Cell Reports. 27(12):3629-3645.e6.. PubMed
  14. Hering L, et al. 2020. Front Immunol. 1.747222222. PubMed
  15. Szodoray P, et al. 2021. Cell Reports. 36(6):109525. PubMed
  16. Lightman SM, et al. 2020. Cell Reports. 1.086805556. PubMed
RRID
AB_2563004 (BioLegend Cat. No. 646403)
AB_2563004 (BioLegend Cat. No. 646404)

Antigen Details

Structure
Molecular weight is approximately 51 kD.
Distribution

IRF4 is reported to be expressed exclusively in the immune system.

Biology Area
Cell Biology, Transcription Factors
Molecular Family
Nuclear Markers
Antigen References

1. Lu R. 2008. Trends Immunol 29:487.

Gene ID
3662 View all products for this Gene ID
UniProt
View information about IRF4 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.

Go To Top Version: 2    Revision Date: 04/21/2022

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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