Alexa Fluor® 594 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
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Product Citations
publications
1_HI30_A594_CD45_Antibody_IHCP_070816
Human paraffin-embedded tonsil tissue slices were prepared with a standard protocol of deparaffination and rehydration. Antigen retrieval was done with Tris-Buffered Saline 20X (1.0M, pH7.4) at 95°C for 40 minutes. The tissue was washed with PBS/ 0.05% Tween20 twice for five minutes and blocked with 5% FBS and 0.2% Gelatin for 30 minutes. Then, the tissue was stained with 5 µg/ml of anti-human CD45 (clone HI30) Alexa Fluor® 594 (red) at 4°C overnight. Nuclei were counterstained with DAPI (blue). The image was captured with a 10X objective.
  • 1_HI30_A594_CD45_Antibody_IHCP_070816
    Human paraffin-embedded tonsil tissue slices were prepared with a standard protocol of deparaffination and rehydration. Antigen retrieval was done with Tris-Buffered Saline 20X (1.0M, pH7.4) at 95°C for 40 minutes. The tissue was washed with PBS/ 0.05% Tween20 twice for five minutes and blocked with 5% FBS and 0.2% Gelatin for 30 minutes. Then, the tissue was stained with 5 µg/ml of anti-human CD45 (clone HI30) Alexa Fluor® 594 (red) at 4°C overnight. Nuclei were counterstained with DAPI (blue). The image was captured with a 10X objective.
  • 19_Human_Liver_Keratin7_CD44_CD45
    Confocal image of human liver sample acquired using the IBEX method of highly multiplexed antibody-based imaging: Cytokeratin 7 (green) in Cycle 1, CD44 (blue) in Cycle 4, and CD45 (magenta) 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® 594 spectral data See high resolution IHC-P data...
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304060 100 µg 248 CHF
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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 (pdf)

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.
Preparation
The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 594 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

IHC-P - Quality tested
SB - Reported in the literature, not verified in house

Recommended Usage

Each lot of this antibody is quality control tested by formalin-fixed paraffin-embedded immunohistochemical staining. For immunohistochemistry, a concentration range of 5.0 - 10 μg/ml is suggested. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 594 has an excitation maximum of 590 nm, and a maximum emission of 617 nm.


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

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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

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. 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. Chou DB, et al. 2020. Nat Biomed Eng. 0.440277778. PubMed
RRID
AB_2629599 (BioLegend Cat. No. 304060)

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

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
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
Go To Top Version: 2    Revision Date: 04.21.2022

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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.

 

BioLegend Inc., 8999 BioLegend Way, San Diego, CA 92121 www.biolegend.com
Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

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.

  • APC anti-human CD45

  • Biotin anti-human CD45

  • FITC anti-human CD45

  • PE anti-human CD45

  • PE/Cyanine5 anti-human CD45

  • Purified anti-human CD45

  • APC/Cyanine7 anti-human CD45

  • PE/Cyanine7 anti-human CD45

  • Alexa Fluor® 488 anti-human CD45

  • Alexa Fluor® 647 anti-human CD45

  • Pacific Blue™ anti-human CD45

  • Alexa Fluor® 700 anti-human CD45

  • PerCP anti-human CD45

  • PerCP/Cyanine5.5 anti-human CD45

  • Brilliant Violet 421™ anti-human CD45

  • Brilliant Violet 570™ anti-human CD45

  • Brilliant Violet 510™ anti-human CD45

  • Brilliant Violet 605™ anti-human CD45

  • Brilliant Violet 650™ anti-human CD45

  • Purified anti-human CD45 (Maxpar® Ready)

  • Brilliant Violet 785™ anti-human CD45

  • Brilliant Violet 711™ anti-human CD45

  • PE/Dazzle™ 594 anti-human CD45

  • Alexa Fluor® 594 anti-human CD45

  • APC/Fire™ 750 anti-human CD45

  • Pacific Blue™ anti-human CD45

  • APC anti-human CD45

  • PE/Cyanine7 anti-human CD45

  • PE/Dazzle™ 594 anti-human CD45

  • TotalSeq™-A0391 anti-human CD45

  • TotalSeq™-B0391 anti-human CD45

  • TotalSeq™-C0391 anti-human CD45

  • PerCP/Cyanine5.5 anti-human CD45

  • APC/Fire™ 750 anti-human CD45

  • PE/Fire™ 640 anti-human CD45

  • APC/Fire™ 810 anti-human CD45

  • Spark YG™ 570 anti-human CD45

  • PE/Fire™ 700 anti-human CD45

  • FITC anti-human CD45

  • PerCP anti-human CD45

  • Alexa Fluor® 660 anti-human CD45 Antibody

  • Spark Violet™ 538 anti-human CD45

  • Spark YG™ 593 anti-human CD45

  • GMP APC/Fire™ 750 anti-human CD45

  • Spark Violet™ 538 anti-human CD45

  • GMP APC anti-human CD45

  • Spark UV™ 387 anti-human CD45

  • GMP Pacific Blue™ anti-human CD45

  • GMP PerCP anti-human CD45

  • GMP FITC anti-human CD45

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