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PerCP/Cyanine5.5 anti-human CD45 Antibody

Pricing & Availability
Clone
HI30 (See other available formats)
Regulatory Status
RUO
Workshop
IV N816
Other Names
LCA, T200
Isotype
Mouse IgG1, κ
Ave. Rating
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Product Citations
publications
HI30_PerCPCyanine55_CD45_Antibody_080818
Human peripheral blood lymphocytes were stained with HI30 PerCP/Cyanine5.5 (filled histogram), or mouse IgG1, κ PerCP/Cyanine5.5 isotype control (open histogram).
  • HI30_PerCPCyanine55_CD45_Antibody_080818
    Human peripheral blood lymphocytes were stained with HI30 PerCP/Cyanine5.5 (filled histogram), or mouse IgG1, κ PerCP/Cyanine5.5 isotype control (open histogram).
  • HI30_PerCPCyanine55_CD45_Antibody_2_101024
    Multiplexed IHC staining of PerCP/Cyanine5.5 anti-CD45 (clone 2D1) on formalin-fixed paraffin-embedded human tonsil tissue, validated for use on the Cellscape™. The tissue was iteratively stained with PerCP/Cyanine5.5 anti-CD45 (clone 2D1, red) and PE anti-CD20 (green) for one hour at room temperature. Nuclei were counterstained with Hoechst 33342. Images were captured with a 20X objective. Scale bar: 50 µm
Compare all formats See PerCP/Cyanine5.5 spectral data
Cat # Size Price Quantity Check Availability Save
304027 25 tests $159
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304028 100 tests $340
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Description

CD45 is a 180-240 kD single chain type I membrane glycoprotein also known as leukocyte common antigen (LCA) and T200. It is a tyrosine phosphatase expressed on the plasma membrane of all hematopoietic cells, except erythrocytes and platelets. CD45 is a signaling molecule that regulates a variety of cellular processes including cell growth, differentiation, cell cycle, and oncogenic transformation. CD45 plays a critical role in T and B cell antigen receptor-mediated activation by dephosphorylating substrates including p56Lck, p59Fyn, and other Src family kinases. CD45 non-covalently associates with lymphocyte phosphatase-associated phosphoprotein (LPAP) on T and B lymphocytes. CD45 has been reported to bind galectin-1 and to be associated with several other cell surface antigens including CD1, CD2, CD3, and CD4.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Human
Reported Reactivity
Chimpanzee
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 PerCP/Cyanine5.5 under optimal conditions.
Concentration
Lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.)
Storage & Handling
The CD45 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 - Community Verified
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.

* PerCP/Cyanine5.5 has a maximum absorption of 482 nm and a maximum emission of 690 nm.

Excitation Laser
Blue Laser (488 nm)
Application Notes

Additional reported applications (for the relevant formats) include: immunohistochemical staining of acetone-fixed frozen tissue sections and formalin-fixed paraffin-embedded tissue sections9, inhibition of CD45 functions4, immunofluorescence11, Western blotting3, and spatial biology (IBEX)16,17.

It was found that the HI30 clone and the 2D1 clone can cross block each other's binding.

Additional Product Notes

For the use of this antibody in spatial biology (SB), we have partnered with Bruker Spatial Biology Biosciences for demonstration of this antibody on their next-generation ChipCytometry instrument called the CellScape™. The CellScape platform is an end-to-end solution for highly multiplexed spatial omics. Combining an advanced, purpose-built imaging system with easy-to-use fluidics for walk-away automation, the CellScape system will accelerate your exploration into the rapidly evolving field of spatial biology. More information on the the Bruker Spatial Biology CellScape and a complete list of our antibodies that have been demonstrated on the instrument can be found here.

Application References

(PubMed link indicates BioLegend citation)
  1. Knapp W, et al. 1989. Leucocyte Typing IV. Oxford University Press. New York.
  2. Kishihara K, et al. 1993. Cell 74:143.
  3. Esser M, et al. 2001. J. Virol. 75:6173. (WB)
  4. Yamada T, et al. 2002. J. Biol. Chem. 277:28830.
  5. Nagano M, et al. 2007. Blood 110:151.
  6. Jiang Q, et al. 2008. Blood 112:2858. PubMed
  7. Morozov A, et al. 2010. Clin Cancer Res. 16:5630. PubMed
  8. Yoshino N, et al. 2000. Exp. Anim. (Tokyo) 49:97. (FC)
  9. Friedman T, et al. 1999. J. Immunol. 162:5256. (IHC)
  10. Oeztuerk-Winder F, et al. 2012. EMBO J. 31:3431. (FC) PubMed
  11. Rees LE, et al. 2003. Clin. Exp. Immunol. 134:497. (IF)
  12. Lee J, et al. 2015. J Exp Med. 212:385. PubMed
  13. Breton G, et al. 2015. J Exp Med. 212:401. PubMed
  14. Marquardt N, et al. 2015. J Immunol. 6:2467. PubMed
  15. Bushway ME, et al. 2014. Biol Reprod. 90(5): 110. (IF) PubMed
  16. Radtke AJ, et al. 2020. Proc Natl Acad Sci USA. 117:33455-33465. (SB) PubMed
  17. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
  1. Duraiswamy J, et al. 2021. Cancer Cell. 39:1623. PubMed
  2. Gur C, et al. 2022. Cell. 185:1373. PubMed
  3. Gao Y, et al. 2023. iScience. 26:106729. PubMed
  4. Changsheng H, et al. 2023. Sci Adv. 9:eade4186. PubMed
  5. Kaczanowska S, et al. 2021. Cell. 184(8):2033-2052.e21. PubMed
  6. Junker F, et al. 2021. Cytometry A. 99:832. PubMed
  7. Jarosch S, et al. 2022. STAR Protoc. 3:101374. PubMed
  8. Zhou L, et al. 2021. MAbs. 13:1964420. PubMed
  9. Jing Li et al. 2018. Immunity. 48(4):773-786 . PubMed
  10. Caggiati A, et al. 2017. Aesthet Surg J. 10.1093/asj/sjw211. PubMed
  11. Gerhauser C et al. 2018. Cancer cell. 34(6):996-1011 . PubMed
  12. Cai D, et al. 2021. Front Mol Biosci. 8:686803. PubMed
  13. Godoy-Pacheco A, et al. 2022. Oncol Lett. 24:288. PubMed
  14. Santos J, et al. 2013. J Transl Med. 11:18. PubMed
  15. Imai Y, et al. 2021. JID Innov. 1:100003. PubMed
  16. Jarosch S, et al. 2021. Cell Rep Methods. 1:100104. PubMed
  17. Schupp J, et al. 2021. Int J Mol Sci. 22:00. PubMed
  18. Evans WS, et al. 2020. Exp Physiol. 105:1408. PubMed
  19. Kanderova V, et al. 2016. Mol Cell Proteomics. 15: 1246-1261. PubMed
  20. Ziegler CGK, et al. 2021. Cell. 184(18):4713-4733.e22. PubMed
  21. Soh KT, et al. 2020. Cytometry B Clin Cytom. 98:161. PubMed
  22. Roug A, et al. 2014. Cytotherapy. 16:392. PubMed
  23. Xu Y, et al. 2021. Cell Reports. 36(6):109524. PubMed
  24. Giraud M, et al. 2012. Proc Natl Acad Sci U S A. 109:535. PubMed
  25. Phansalkar R, et al. 2021. Elife. 10:. PubMed
  26. Lavaert M, et al. 2020. Frontiers in Immunology. 1.610416667. PubMed
  27. Johnson AJ, et al. 2021. Cancer Immunol Res. 9:1047. PubMed
RRID
AB_893338 (BioLegend Cat. No. 304027)
AB_893338 (BioLegend Cat. No. 304028)

Antigen Details

Structure
Tyrosine phosphatases, type I transmembrane protein, 180-240 kD (multiple isoforms)
Distribution

Hematopoietic cells, not expressed in circulating erythrocytes or platelets

Function
TCR and BCR mediated activation
Ligand/Receptor
Galectin-1, CD2, CD3, CD4
Cell Type
Hematopoietic stem and progenitors, Mesenchymal Stem Cells
Biology Area
Cell Biology, Immunology, Inhibitory Molecules, Innate Immunity, Neuroscience, Neuroscience Cell Markers, Stem Cells
Molecular Family
CD Molecules
Antigen References
  1. Thomas M. 1989. Annu. Rev. Immunol. 7:339.
  2. Trowbridge I, et al. 1994. Annu. Rev. Immunol.12:85.
Gene ID
5788 View all products for this Gene ID
UniProt
View information about CD45 on UniProt.org

Related FAQs

How stable is PerCP/Cyanine5.5 tandem as compared to PerCP alone?

PerCP/Cyanine5.5 is quite photostable and also better than PerCP alone in withstanding fixation.

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 CD45 Reagents Request Custom Conjugation
Description Clone Applications
APC anti-human CD45 HI30 FC
Biotin anti-human CD45 HI30 FC
FITC anti-human CD45 HI30 FC
PE anti-human CD45 HI30 FC
PE/Cyanine5 anti-human CD45 HI30 FC
Purified anti-human CD45 HI30 FC,CyTOF®,IHC-P,WB,IHC-F,ICC
APC/Cyanine7 anti-human CD45 HI30 FC
PE/Cyanine7 anti-human CD45 HI30 FC
Alexa Fluor® 488 anti-human CD45 HI30 FC
Alexa Fluor® 647 anti-human CD45 HI30 FC
Pacific Blue™ anti-human CD45 HI30 FC
Alexa Fluor® 700 anti-human CD45 HI30 FC
PerCP anti-human CD45 HI30 FC
PerCP/Cyanine5.5 anti-human CD45 HI30 FC,SB
Brilliant Violet 421™ anti-human CD45 HI30 FC
Brilliant Violet 570™ anti-human CD45 HI30 FC
Brilliant Violet 510™ anti-human CD45 HI30 FC
Brilliant Violet 605™ anti-human CD45 HI30 FC
Brilliant Violet 650™ anti-human CD45 HI30 FC
Purified anti-human CD45 (Maxpar® Ready) HI30 FC,CyTOF®
Brilliant Violet 785™ anti-human CD45 HI30 FC
Brilliant Violet 711™ anti-human CD45 HI30 FC
PE/Dazzle™ 594 anti-human CD45 HI30 FC
Alexa Fluor® 594 anti-human CD45 HI30 IHC-P,SB
APC/Fire™ 750 anti-human CD45 HI30 FC
Pacific Blue™ anti-human CD45 HI30 FC
APC anti-human CD45 HI30 FC
PE/Cyanine7 anti-human CD45 HI30 FC
PE/Dazzle™ 594 anti-human CD45 HI30 FC
TotalSeq™-A0391 anti-human CD45 HI30 PG
TotalSeq™-B0391 anti-human CD45 HI30 PG
TotalSeq™-C0391 anti-human CD45 HI30 PG
PerCP/Cyanine5.5 anti-human CD45 HI30 FC
APC/Fire™ 750 anti-human CD45 HI30 FC
PE/Fire™ 640 anti-human CD45 HI30 FC
APC/Fire™ 810 anti-human CD45 HI30 FC
Spark YG™ 570 anti-human CD45 HI30 IHC-P
PE/Fire™ 700 anti-human CD45 HI30 FC
FITC anti-human CD45 HI30 FC
PerCP anti-human CD45 HI30 FC
Alexa Fluor® 660 anti-human CD45 Antibody HI30 FC
Spark Violet™ 538 anti-human CD45 HI30 FC
Spark YG™ 593 anti-human CD45 HI30 FC
GMP APC/Fire™ 750 anti-human CD45 HI30 FC
Spark Violet™ 538 anti-human CD45 HI30 FC
GMP APC anti-human CD45 HI30 FC
Spark UV™ 387 anti-human CD45 HI30 FC
GMP Pacific Blue™ anti-human CD45 HI30 FC
GMP PerCP anti-human CD45 HI30 FC
GMP FITC anti-human CD45 HI30 FC
PE anti-human CD45 HI30 FC
GMP PE/Dazzle™ 594 anti-human CD45 HI30 FC
GMP PerCP/Cyanine5.5 anti-human CD45 HI30 FC
Spark Blue™ 515 anti-human CD45 HI30 FC
TotalSeq™-D0391 anti-human CD45 HI30 PG
GMP PE/Cyanine7 anti-human CD45 HI30 FC
Spark Violet™ 500 anti-human CD45 HI30 FC
GMP PE anti-human CD45 HI30 FC
PE/Fire™ 810 anti-human CD45 HI30 FC
Spark PLUS UV395™ anti-human CD45 HI30 FC
Spark NIR™ 685 anti-human CD45 HI30 FC
Spark Violet™ 423 anti-human CD45 HI30 FC
Go To Top Version: 4    Revision Date: 10/10/2024

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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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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