Alexa Fluor® 647 anti-mouse CD45 Antibody

Pricing & Availability
Clone
30-F11 (See other available formats)
Regulatory Status
RUO
Other Names
T200, Ly-5, LCA
Isotype
Rat IgG2b, κ
Ave. Rating
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Product Citations
publications
56_Mouse_Lymph_Node_CD45
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: CD45 (magenta) 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).
  • 56_Mouse_Lymph_Node_CD45
    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: CD45 (magenta) 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).
  • a)30-F11_Alx647_090507
    C57BL/6 mouse splenocytes stained with 30-F11 Alexa Fluor® 647
  • b)30-F11_A647_CD45_Antibody_ICC_031520
    C57 mouse bone marrow cells were fixed with 2% paraformaldehyde (PFA), and then stained with 10 µg/ml of CD11b (clone M1/70) Brilliant Violet 510™ (red) and 10 µg/ml of CD45 (clone 30-F11) Alexa Fluor® 647 (blue) for 30 minutes at room temperature. The image was captured by 60X objective.
  • c)30-F11_A647_CD45_Antibody_3D_IHC_05112021.png
    Paraformaldehyde-fixed (4%), mouse intestine tissue section was processed according to the Ce3DTM Tissue Clearing Kit protocol (cat. no. 427701). The section was costained with anti-Tubulin β 3 (TUBB3) Antibody (clone TUJ1) Alexa Fluor® 594 at 5 µg/mL (green), and anti-mouse CD45 Antibody (clone 30-F11) Alexa Fluor® 647 at 5 µg/mL (magenta). The section was then optically cleared and mounted in a sample chamber. The image was captured with a 20X objective using Zeiss 780 confocal microscope and processed by Imaris image analysis software.
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103123 25 µg £70
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103124 100 µg £157
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Description

CD45 is a 180-240 kD glycoprotein also known as the leukocyte common antigen (LCA), T200, or Ly-5. It is a member of the protein tyrosine phosphatase (PTP) family, expressed on all hematopoietic cells except mature erythrocytes and platelets. There are different isoforms of CD45 that arise from variable splicing of exons 4, 5, and 6, which encode A, B, and C determinants, respectively. CD45 plays a key role in TCR and BCR signal transduction. These isoforms are very specific to the activation and maturation state of the cell as well as cell type. The primary ligands for CD45 are galectin-1, CD2, CD3, CD4, TCR, CD22, and Thy-1.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Mouse
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Mouse thymus or spleen
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® 647 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
ICC, 3D IHC - Verified
SB - Community verified
IHC, 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 ≤ 0.25 µg per 106 cells in 100 µl volume. For 3D immunohistochemistry on formalin-fixed tissues, a concentration of 5.0 µg/mL is suggested. It is recommended that the reagent be titrated for optimal performance for other applications.

* Alexa Fluor® 647 has a maximum emission of 668 nm when it is excited at 633nm / 635nm.


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

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Excitation Laser
Red Laser (633 nm)
Application Notes

Clone 30-F11 reacts with all isoforms and both CD45.1 and CD45.2 alloantigens of CD45.

Additional reported applications (for relevant formats) include: immunoprecipitation3, complement-dependent cytotoxicity1,5, immunohistochemistry (acetone-fixed frozen sections, zinc-fixed paraffin-embedded sections and formalin-fixed paraffin-embedded sections)4,6, Western blotting7, and spatial biology (IBEX)10,11. The Ultra-LEAF™ purified antibody (Endotoxin < 0.01 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays (Cat. No. 103163 and 103164).

Additional Product Notes

For use in spatial biology, this antibody has been demonstrated for use in immunohistochemistry using IBEX (Reported in the literature, not verified in house) and the NanoString GeoMx® Digital Spatial Profiler.

IBEX: 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).

NanoString GeoMx®: This product has been verified for IHC-F (Immunohistochemistry - frozen tissue sections) on the NanoString GeoMx® Digital Spatial Profiler. The GeoMx® enables researchers to perform spatial analysis of protein and RNA targets in FFPE and fresh frozen human and mouse samples. For more information about our spatial biology products and the GeoMx® platform, please visit our spatial biology page.

Application References

(PubMed link indicates BioLegend citation)
  1. Podd BS, et al. 2006. J. Immunol. 176:6532. (FC, CMCD) PubMed
  2. Haynes NM, et al. 2007. J. Immunol. 179:5099. (FC)
  3. Ledbetter JA, et al. 1979. Immunol. Rev. 47:63. (IP)
  4. Simon DI, et al. 2000. J. Clin. Invest. 105:293. (IHC)
  5. Seaman WE. 1983. J. Immunol. 130:1713. (CMCD)
  6. Cornet A, et al. 2001. P. Natl. Acad. Sci. USA 98:13306. (IHC)
  7. Tsuboi S and Fukuda M. 1998. J. Biol. Chem. 273:30680. (WB) PubMed
  8. Liu F, et al. 2012. Blood. 119:3295. PubMed
  9. Pelletier AN, et al. 2012. J. Immunol. 188:5561. 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. Yan Y, et al. 2021. Immunity. 54:499. PubMed
  2. Osterloh A, et al. 2016. Infect Immun . . PubMed
  3. Moderzynski K, et al. 2016. PLoS Negl Trop Dis. . PubMed
  4. Boal-Carvalho I, et al. 2020. EMBO Rep. 21:e50421. PubMed
  5. Cho J, et al. 2016. Nat Commun. 7:13373. PubMed
  6. Donato C, et al. 2020. Cell Reports. 32(10):108105. PubMed
  7. Hu X, et al. 2016. Nat Commun. 7:13095. PubMed
  8. Moore SM, et al. 2019. Transl Psychiatry. 9:24. PubMed
  9. Biffi G, et al. 2018. Cancer Discov. 2:282. PubMed
  10. Hu K, et al. 2015. PLoS One. 10: 0137123. PubMed
  11. Perner C, et al. 2020. Immunity. 53(5):1063-1077.e7. PubMed
  12. Sutton NR, et al. 2019. Arterioscler Thromb Vasc Biol. 40:61. PubMed
  13. Lin JR et al. 2018. eLife. 7 pii: e31657. PubMed
  14. Fang EF, et al. 2019. Nat Neurosci. 22:401. PubMed
  15. Uceda-Castro R, et al. 2022. Cell Rep Med. 3:100821. PubMed
  16. Ruhland MK, et al. 2020. Cancer Cell. 37(6):786-799.e5. PubMed
  17. Piedrafita G, et al. 2020. Nat Commun. 11:1429. PubMed
  18. Donato C, et al. 2021. STAR Protocols. 2(2):100480. PubMed
  19. Pineda CM, et al. 2019. J Cell Biol. 218:3212. PubMed
  20. Kim S, et al. 2022. Redox Biol. 54:102347. PubMed
  21. Kang MH, et al. 2020. Nat Commun. 3.186805556. PubMed
  22. Kuhn JA, et al. 2021. Elife. 10:. PubMed
  23. Levesque S, et al. 2019. Oncoimmunology. 8:e1657375. PubMed
  24. Simões FC, et al. 2020. Nat Commun. 0.875. PubMed
  25. Tummers B, et al. 2020. Immunity. 52(6):994-1006.e8. PubMed
  26. Huang L, et al. 2018. Exp Neurol. 300:41. PubMed
  27. Sakamoto K, et al. 2021. Immunity. 54:2321. PubMed
  28. Wang S, et al. 2021. Commun Biol. 22:. PubMed
  29. Shibuya M, et al. 2021. iScience. 24:103131. PubMed
  30. de Winde CM, et al. 2021. J Cell Sci. 134: . PubMed
  31. Lu YJ, et al. 2021. Cell Rep. 36:109696. PubMed
  32. Ovadya Y, et al. 2018. Nat Commun. 9:5435. PubMed
  33. Yomtoubian S, et al. 2020. Cell Reports. 30(3):755-770.e6.. PubMed
  34. Thacker VV, et al. 2020. Elife. 9:00. PubMed
  35. Sanders K, et al. 2015. Cancer Immunol Res. 3: 891-901. PubMed
  36. Clemente–Casares X, et al. 2017. Immunity. 47:974. PubMed
  37. Bakalar MH et al. 2018. Cell. 174(1):131-142 . PubMed
  38. Chen W, et al. 2016. Nat Commun. 7: 11302. PubMed
  39. Xiong J et al. 2018. Molecular cell. 69(4):689-698 . PubMed
  40. Williams IM, et al. 2018. J Clin Invest. 128:699. PubMed
  41. Koyama M et al. 2019. Immunity. 51(5):885-898 . PubMed
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  43. Oni TE, et al. 2020. J Exp Med. :217. PubMed
RRID
AB_493534 (BioLegend Cat. No. 103123)
AB_493533 (BioLegend Cat. No. 103124)

Antigen Details

Structure
Protein tyrosine phosphatase (PTP) family, 180-240 kD
Distribution

All hematopoietic cells except mature erythrocytes and platelets

Function
Phosphatase, T and B cell activation
Ligand/Receptor
Galectin-1, CD2, CD3, CD4, TCR, CD22, Thy-1
Cell Type
B cells, Dendritic cells, Mesenchymal Stem Cells, Tregs
Biology Area
Cell Biology, Immunology, Inhibitory Molecules, Innate Immunity, Neuroscience, Neuroscience Cell Markers, Stem Cells
Molecular Family
CD Molecules
Antigen References

1. Barclay A, et al. 1997. The Leukocyte Antigen FactsBook Academic Press.
2. Trowbridge IS, et al. 1993. Annu. Rev. Immunol. 12:85.
3. Kishihara K, et al. 1993. Cell 74:143.
4. Pulido R, et al. 1988. J. Immunol. 140:3851.

Gene ID
19264 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-mouse CD45 30-F11 FC
Biotin anti-mouse CD45 30-F11 FC
FITC anti-mouse CD45 30-F11 FC
PE anti-mouse CD45 30-F11 FC
PE/Cyanine5 anti-mouse CD45 30-F11 FC
Purified anti-mouse CD45 30-F11 FC,IHC-F,CyTOF®,IP,CMCD,IHC,WB
PE/Cyanine7 anti-mouse CD45 30-F11 FC
APC/Cyanine7 anti-mouse CD45 30-F11 FC
Alexa Fluor® 488 anti-mouse CD45 30-F11 FC,SB
Alexa Fluor® 647 anti-mouse CD45 30-F11 FC,ICC,IHC,3D IHC,SB
Pacific Blue™ anti-mouse CD45 30-F11 FC
Alexa Fluor® 700 anti-mouse CD45 30-F11 FC,SB
PerCP/Cyanine5.5 anti-mouse CD45 30-F11 FC
PerCP anti-mouse CD45 30-F11 FC
Alexa Fluor® 594 anti-mouse CD45 30-F11 IHC-F,FC,3D IHC
Brilliant Violet 421™ anti-mouse CD45 30-F11 FC,SB
Brilliant Violet 570™ anti-mouse CD45 30-F11 FC
Brilliant Violet 510™ anti-mouse CD45 30-F11 FC
Brilliant Violet 605™ anti-mouse CD45 30-F11 FC
Purified anti-mouse CD45 (Maxpar® Ready) 30-F11 FC,CyTOF®
PE/Dazzle™ 594 anti-mouse CD45 30-F11 FC
Brilliant Violet 711™ anti-mouse CD45 30-F11 FC
Brilliant Violet 785™ anti-mouse CD45 30-F11 FC
Brilliant Violet 650™ anti-mouse CD45 30-F11 FC
APC/Fire™ 750 anti-mouse CD45 30-F11 FC
Brilliant Violet 750™ anti-mouse CD45 30-F11 FC
TotalSeq™-A0096 anti-mouse CD45 30-F11 PG
TotalSeq™-B0096 anti-mouse CD45 30-F11 PG
Ultra-LEAF™ Purified anti-mouse CD45 30-F11 FC,CyTOF®,IP,CMCD,IHC,WB
Spark Blue™ 550 anti-mouse CD45 30-F11 FC
Spark NIR™ 685 anti-mouse CD45 30-F11 FC
TotalSeq™-C0096 anti-mouse CD45 30-F11 PG
Spark YG™ 570 anti-mouse CD45 30-F11 IHC-F
PE/Fire™ 640 anti-mouse CD45 30-F11 FC
APC/Fire™ 810 anti-mouse CD45 30-F11 FC
PE/Fire™ 700 anti-mouse CD45 30-F11 FC
Spark Violet™ 538 anti-mouse CD45 30-F11 FC
Spark YG™ 593 anti-mouse CD45 30-F11 FC
Spark Blue™ 574 anti-mouse CD45 Antibody 30-F11 FC
Spark UV™ 387 anti-mouse CD45 30-F11 FC
Spark Red™ 718 anti-mouse CD45 (Flexi-Fluor™) 30-F11 FC
Go To Top Version: 7    Revision Date: 01/31/2024

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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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