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Alexa Fluor® 488 anti-human HLA-DR Antibody

Pricing & Availability
Clone
L243 (See other available formats)
Regulatory Status
RUO
Other Names
Major Histocompatibility Class II, MHC class II
Isotype
Mouse IgG2a, κ
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Product Citations
publications
1_L243_Alx488_072607
Human peripheral blood lymphocytes stained with L243 Alexa Fluor® 488
  • 1_L243_Alx488_072607
    Human peripheral blood lymphocytes stained with L243 Alexa Fluor® 488
  • 2_4_Human_LN_CD69_HLA-DR
    Confocal image of human lymph node sample acquired using the IBEX method of highly multiplexed antibody-based imaging: HLA-DR (cyan) in Cycle 3, CD69 (red) 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).
  • 3_9_Human_LN_HLA-DR_Va72
    Confocal image of human lymph node sample acquired using the IBEX method of highly multiplexed antibody-based imaging: HLA-DR (purple) in Cycle 3 and Vα7.2 (yellow) 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).
  • 4_20_Human_Liver_HLADR_Vimentin
    Confocal image of human liver sample acquired using the IBEX method of highly multiplexed antibody-based imaging: HLA-DR (cyan) in Cycle 2 and Vimentin (red) in Cycle 4. 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).
Compare all formats See Alexa Fluor® 488 spectral data
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307619 25 tests $100
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307620 100 tests $240
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307656 100 µg $258
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Description

HLA-DR is a heterodimeric cell surface glycoprotein comprised of a 36 kD α (heavy) chain and a 27 kD β (light) chain. It is expressed on B cells, activated T cells, monocytes/macrophages, dendritic cells, and other non-professional APCs. In conjunction with the CD3/TCR complex and CD4 molecules, HLA-DR is critical for efficient peptide presentation to CD4+ T cells.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Human, Cynomolgus, Rhesus
Reported Reactivity
African Green, Baboon, Chimpanzee, Dog, Common Marmoset, Squirrel Monkey, Cotton-topped Tamarin
Antibody Type
Monoclonal
Host Species
Mouse
Formulation
µg size: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
test sizes: 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
µg sizes: 0.5 mg/mL
test sizes: lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.)
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 using the µg size, the suggested use of this reagent is ≤1.0 µg per million cells in 100 µl volume. For flow cytometric staining using test sizes, 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

The L243 monoclonal antibody reacts with the HLA-DR antigen, a member of MHC class II molecules. It does not cross react with HLA-DP and HLA-DQ. Clone L243 binds a conformational epitope on HLA-DRa which depends on the correct folding of the aß heterodimer.19

Additional reported applications (for the relevant formats) include: immunoprecipitation8, Western blotting8, in vitro blocking of mixed lymphocyte reactions9,10, depeletion of MHC class II cells7, immunohistochemical staining of acetone-fixed frozen sections4,5, and spatial biology (IBEX)21,22. For sensitive functional assays, we recommend using the Ultra-LEAF™ purified antibody (Endotoxin < 0.01 EU/µg, Azide-Free, 0.2 µm filtered) (Cat. No. 307648, 307665 - 307669).

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. Brodsky F. 1984. Immunogenetics 19:179.
  2. Robbins P, et al. 1987. Human Immunol. 18:301.
  3. Stites D, et al. 1986. Clin. Immunol. Immunopathol. 38:161.
  4. Warnke R, et al. 1980. J. Histochem. Cytochem. 28:771. (IHC)
  5. Engleman E, et al. 1981. P. Natl. Acad. Sci. USA 78:1791. (IHC)
  6. Zipf T, et al. 1981. Cancer Res. 41:4786.
  7. Goodier M, et al. 2000. J. Immunol. 165:139. (Depletion)
  8. Esser M, et al. 2001. J. Virol. 75:6173. (IP, WB)
  9. Kalka-Moll WM, et al. 2002. J. Immunol. 169:6149. (Block)
  10. Wang RF, et al. 1999. Science 284:1351. (Block)
  11. Zaba LC, et al. 2007. J. Exp. Med. 204:3183. PubMed
  12. Fujita H, et al. 2009. P. Natl. Acad. Sci. USA 106:21795. PubMed
  13. Charles N, et al. 2010. Nat. Med. 16:701. (FC) PubMed
  14. Goncalves RM, et al. 2010. Infect. Immun. 78:4763. PubMed
  15. Yoshino N, et al. 2000. Exp. Anim. (Tokyo) 49:97. (FC)
  16. Kim WK, et al. 2006. Am. J. Pathol. 168:822. (FC)
  17. Stein R, et al. 2011. Leuk. Lymphoma 52:273.
  18. Galkowska H, et al. 1996. Vet. Immunol. Immunopathol. 53:329.
  19. Moro M, et al. 2005. BMC Immunol. 6:24.
  20. Lauterbach N, et al. 2014. Mol Immunol. 59:19. PubMed
  21. Radtke AJ, et al. 2020. Proc Natl Acad Sci USA. 117:33455-33465. (SB) PubMed
  22. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
  1. Balmert SC, et al. 2022. iScience. 25:105045. PubMed
  2. Smith MH, et al. 2023. Nat Immunol. . PubMed
  3. Roberts A, et al. 2021. Sci Rep. 11:4030. PubMed
  4. Casasola-LaMacchia A, et al. 2021. Sci Rep. 11:1028. PubMed
  5. Lamichhane R, et al. 2020. Eur J Immunol. 50:178. PubMed
  6. Lindesmith LC, et al. 2020. Cell Mol Gastroenterol Hepatol. 0.586805556. PubMed
  7. Kim ST, et al. 2022. Nat Commun. 13:1970. PubMed
  8. Nazitto R, et al. 2021. J Immunol. 206:2949. PubMed
  9. Forbester JL, et al. 2020. J Virol. 94:. PubMed
  10. Salvi V, et al. 2018. JCI Insight. 3. PubMed
  11. Savage AK, et al. 2021. iScience. 24(5):102404. PubMed
  12. Chung SH, et al. 2021. Hum Gene Ther. 32:682. PubMed
  13. Walseng E, et al. 2008. J Biol Chem. 283:14717. PubMed
RRID
AB_493175 (BioLegend Cat. No. 307619)
AB_493175 (BioLegend Cat. No. 307620)
AB_493175 (BioLegend Cat. No. 307656)

Antigen Details

Structure
Ig superfamily, MHC class II, heterodimeric transmembrane protein, 36 kD heavy and 27 kD light chain
Distribution

B cells, activated T cells, monocytes/macrophages, dendritic cells, other APCs

Function
Peptide presentation
Ligand/Receptor
CD3/TCR, CD4
Cell Type
Antigen-presenting cells, B cells, Dendritic cells, Macrophages, Monocytes, T cells, Tregs
Biology Area
Immunology, Innate Immunity
Molecular Family
MHC Antigens
Antigen References

1. Levacher M, et al. 1990. Clin. Exp. Immunol. 81:177.
2. Terstappen L, et al. 1990. J. Leukocyte Biol. 48:138.
3. Edwards JA, et al. 1986. J. Immunol. 137:490.
4. van Es A, et al. 1984. Transplantation 37:65.
5. O'Doherty U, et al. 1994. Immunology 82:487.
6. Thomas R, et al. 1994. J. Immunol. 153:4016.
7. Grouard G, et al. 1996. Nature 384:364.

Gene ID
3122 View all products for this Gene ID 3123 View all products for this Gene ID
UniProt
View information about HLA-DR 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.

Other Formats

View All HLA-DR Reagents Request Custom Conjugation
Description Clone Applications
APC anti-human HLA-DR L243 FC
FITC anti-human HLA-DR L243 FC
PE anti-human HLA-DR L243 FC
PE/Cyanine5 anti-human HLA-DR L243 FC
Purified anti-human HLA-DR L243 FC,CyTOF®,IP,WB,Block,IHC-F
Biotin anti-human HLA-DR L243 FC
PE/Cyanine7 anti-human HLA-DR L243 FC
APC/Cyanine7 anti-human HLA-DR L243 FC
Alexa Fluor® 488 anti-human HLA-DR L243 FC,SB
Alexa Fluor® 647 anti-human HLA-DR L243 FC
Pacific Blue™ anti-human HLA-DR L243 FC
Alexa Fluor® 700 anti-human HLA-DR L243 FC
PerCP anti-human HLA-DR L243 FC
PerCP/Cyanine5.5 anti-human HLA-DR L243 FC
Brilliant Violet 605™ anti-human HLA-DR L243 FC
Brilliant Violet 421™ anti-human HLA-DR L243 FC
Brilliant Violet 570™ anti-human HLA-DR L243 FC
Brilliant Violet 711™ anti-human HLA-DR L243 FC
Brilliant Violet 785™ anti-human HLA-DR L243 FC
Brilliant Violet 510™ anti-human HLA-DR L243 FC
Ultra-LEAF™ Purified anti-human HLA-DR L243 FC,CyTOF®,IP,WB,Block,IHC-F
Brilliant Violet 650™ anti-human HLA-DR L243 FC
Purified anti-human HLA-DR (Maxpar® Ready) L243 FC,CyTOF®
PE/Dazzle™ 594 anti-human HLA-DR L243 FC
FITC anti-human HLA-DR L243 FC
APC/Fire™ 750 anti-human HLA-DR L243 FC
Pacific Blue™ anti-human HLA-DR L243 FC
APC anti-human HLA-DR L243 FC
PE/Dazzle™ 594 anti-human HLA-DR L243 FC
PE/Cyanine7 anti-human HLA-DR L243 FC
TotalSeq™-A0159 anti-human HLA-DR L243 PG
TotalSeq™-B0159 anti-human HLA-DR L243 PG
TotalSeq™-C0159 anti-human HLA-DR L243 PG
Brilliant Violet 750™ anti-human HLA-DR L243 FC
APC/Fire™ 750 anti-human HLA-DR L243 FC
PerCP/Cyanine5.5 anti-human HLA-DR L243 FC
APC/Fire™ 810 anti-human HLA-DR L243 FC
PE/Fire™ 640 anti-human HLA-DR L243 FC
PE anti-human HLA-DR L243 FC
Spark Violet™ 538 anti-human HLA-DR Antibody L243 FC
KIRAVIA Blue 520™ anti-human HLA-DR L243 FC
TotalSeq™-D0159 anti-human HLA-DR L243 PG
PE/Fire™ 810 anti-human HLA-DR L243 FC
GMP PE/Dazzle™ 594 anti-human HLA-DR L243 FC
Spark Violet™ 423 anti-human HLA-DR L243 FC
PerCP anti-human HLA-DR L243 FC
GMP FITC anti-human HLA-DR L243 FC
GMP APC anti-human HLA-DR L243 FC
GMP PE/Cyanine7 anti-human HLA-DR L243 FC
GMP Pacific Blue™ anti-human HLA-DR L243 FC
GMP APC/Fire™ 750 anti-human HLA-DR L243 FC
Spark Violet™ 500 anti-human HLA-DR L243 FC
GMP PerCP/Cyanine5.5 anti-human HLA-DR L243 FC
GMP PE anti-human HLA-DR L243 FC
Spark UV™ 387 anti-human HLA-DR L243 FC
Spark Blue™ 515 anti-human HLA-DR L243 FC
Spark NIR™ 685 anti-human HLA-DR L243 FC
PerCP/Fire™ 806 anti-human HLA-DR L243 FC
PE/Fire™ 700 anti-human HLA-DR L243 FC
Spark Blue™ 550 anti-human HLA-DR (Flexi-Fluor™) L243 FC
Spark Red™ 718 anti-human HLA-DR (Flexi-Fluor™) L243 FC
PE/Fire™ 744 anti-human HLA-DR L243 FC
Spark PLUS UV395™ anti-human HLA-DR L243 FC
Spark Violet™ 423 anti-human HLA-DR L243 FC
Go To Top Version: 5    Revision Date: 08/26/2022

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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