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Alexa Fluor® 594 anti-mouse/human CD44 Antibody

Pricing & Availability
Clone
IM7 (See other available formats)
Regulatory Status
RUO
Other Names
Hermes, Pgp-1, H-CAM, HUTCH-1, ECMR III, gp85, Ly-24
Isotype
Rat IgG2b, κ
Ave. Rating
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Product Citations
publications
IM7_AF594_CD44_Antibody_1_IF_111416
bEND3 cells were fixed and blocked according to standard immunofluorescence protocols. Cells were stained with CD44 (clone IM7) Alexa Fluor® 594 and DAPI (blue).
  • IM7_AF594_CD44_Antibody_1_IF_111416
    bEND3 cells were fixed and blocked according to standard immunofluorescence protocols. Cells were stained with CD44 (clone IM7) Alexa Fluor® 594 and DAPI (blue).
  • IM7_AF594_CD44_Antibody_2_FC_111416
    C57BL/6 mouse splenocytes were stained with CD44 (clone IM7) Alexa Fluor® 594 (filled histogram). The data was acquired by BD LSRFortessa™ cell analyzer equipped with Yellow-Green Laser (561 nm).
  • IM7_AF594_CD44_Antibody_3_IHCP_111416
    Human paraffin-embedded intestine tissue was prepared with standard deparaffination, rehydration, antigen retrieval and blocking protocols. Tissue was stained with anti-human CD44 (clone IM7) Alexa Fluor® 594 (red) and DAPI (blue). See additional supplemental data for detailed information.
Compare all formats See Alexa Fluor® 594 spectral data See high resolution IF/IHC-P data...
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103054 100 µg 253 CHF
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Description

CD44 is a 80-95 kD glycoprotein also known as Hermes, Pgp1, H-CAM, or HUTCH. It is expressed on all leukocytes, endothelial cells, hepatocytes, and mesenchymal cells. As B and T cells become activated or progress to the memory stage, CD44 expression increases from low or mid levels to high levels. Thus, CD44 has been reported to be a valuable marker for memory cell subsets. High CD44 expression on Treg cells has been associated with potent suppressive function via high production of IL-10. CD44 is an adhesion molecule involved in leukocyte attachment to and rolling on endothelial cells, homing to peripheral lymphoid organs and to the sites of inflammation, and leukocyte aggregation.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Mouse, Human
Reported Reactivity
Chimpanzee, Baboon, Cynomolgus, Rhesus, Squirrel Monkey, Horse, Cow, Pig, Dog, Cat
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Dexamethasone-induced myeloid leukemia M1 cells
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

IF - Quality tested
FC, IHC-P - Verified
SB - Reported in the literature, not verified in house
Recommended Usage

Each lot of this antibody is quality control tested by immunofluorescence staining. For immunofluorescence microscopy, a concentration range of 2.5-10 μg/ml is recommended. For flow cytometric staining, the suggested use of this reagent is ≤0.25 µg per million cells in 100 µl volume. For immunohistochemical staining on formalin-fixed paraffin-embedded tissue sections, a concentration range of 5-10 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

Clone IM7 has been reported to recognize an epitope common to alloantigens and all isoforms of CD4417,18 that is located between amino acids 145 and 18620. This clone has been verified for immunocytochemistry (ICC) and frozen immunohistochemistry (IHC-F). Additional reported applications (for the relevant formats) include: immunohistochemistry of acetone-fixed frozen sections and formalin-fixed paraffin-embedded sections6,7, complement-mediated cytotoxicity1, immunoprecipitation1,3, in vivo inhibition of DTH4,5, and spatial biology (IBEX)23,24. The Ultra-LEAF™ purified antibody (Endotoxin < 0.01 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays (Cat. No. 103046, 103065 - 103069).

Cross-reactivity to ferret has been reported by a collaborator, but not verified in house.

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. Trowbridge IS, et al. 1982. Immunogenetics 15:299. (ICFC, IP, CMCD)
  2. Katoh S, et al. 1994. J. Immunol. 153:3440. (ELISA)
  3. Budd RC, et al. 1987. J. Immunol. 138:3120. (IP)
  4. Camp RL, et al. 1993. J. Exp. Med. 178:497. (Block)
  5. Weiss JM, et al. 1997. J. Cell Biol. 137:1137. (Block)
  6. Frank NY, et al. 2005. Cancer Res. 65:4320. (IHC) PubMed
  7. Cuff CA, et al. 2001. J. Clin. Invest. 108:1031. (IHC)
  8. Lee JW, et al. 2006. Nature Immunol. 8:181.
  9. Zhang N, et al. 2005. J. Immunol. 174:6967. PubMed
  10. Huabiao C, et al. 2005. J. Immunol. 175:591. PubMed
  11. Gui J, et al. 2007. Int. Immunol. 19:1201. PubMed
  12. Wang XY, et al. 2008. Blood 111:2436. PubMed
  13. Kenna TJ, et al. 2008. Blood 111:2091. PubMed
  14. Yamazaki J, et al. 2009. Blood PubMed
  15. Kmieciak M, et al. 2009. J. Transl. Med. 7:89. (FC) PubMed
  16. Chen YW, et al. 2010. Mol. Cancer Ther. 9:2879. PubMed
  17. Zheng Z, et al. 1995. J. Cell. Biol. 130:485.
  18. Wiranowska M, et al. 2010. Int. J. Cancer 127:532.
  19. Hirokawa Y, et al. 2014. Am J Physiol Gastrointerest Liver Physiol. 306:547. PubMed
  20. Sandmaier BM, et al. 1998. Blood 91:3494.
  21. Yang Y, et al. 2015. Hypertension. 65:1047. PubMed
  22. Peterson VM, et al. 2017. Nat. Biotechnol. 35:936. (PG)
  23. Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed
  24. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
  1. Campisi L, et al. 2022. Nature. 606:945. PubMed
  2. Puigdelloses M, et al. 2021. J Immunother Cancer. 9:. PubMed
RRID
AB_2563183 (BioLegend Cat. No. 103054)

Antigen Details

Structure
Variable splicing of CD44 gene generates many CD44 isoforms, 80-95 kD
Distribution

All leukocytes, epithelial cells, endothelial cells, hepatocytes, mesenchymal cells

Function
Leukocyte attachment and rolling on endothelial cells, stromal cells and ECM
Ligand/Receptor
Hyaluronan, MIP-1β, fibronectin, collagen
Cell Type
B cells, Endothelial cells, Epithelial cells, Leukocytes, Mesenchymal cells, Mesenchymal Stem Cells, Tregs
Biology Area
Cell Adhesion, Cell Biology, Immunology, Stem Cells
Molecular Family
Adhesion Molecules, CD Molecules
Antigen References

1. Barclay AN, et al. 1997. The Leukocyte Antigen FactsBook Academic Press.
2. Haynes BF, et al. 1991. Cancer Cells 3:347.
3. Goldstein LA, et al. 1989. Cell 56:1063.
4. Mikecz K, et al. 1995. Nat. Med. 1:558.
5. Hegde V, et al. 2008. J. Leukocyte Biol. 84:134.
6. Liu T, et al. 2009. Biol. Direct 4:40.

Gene ID
12505 View all products for this Gene ID 960 View all products for this Gene ID
UniProt
View information about CD44 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 CD44 Reagents Request Custom Conjugation
Description Clone Applications
APC anti-mouse/human CD44 IM7 FC
Biotin anti-mouse/human CD44 IM7 FC,ICC,IHC-P
FITC anti-mouse/human CD44 IM7 FC,ICC,IHC-P
PE/Cyanine5 anti-mouse/human CD44 IM7 FC
Purified anti-mouse/human CD44 IM7 FC,IHC-F,CyTOF®,ELISA,ICC,IHC-P,IP,CMCD,Stim
Brilliant Violet 605™ anti-mouse/human CD44 IM7 FC
PE anti-mouse/human CD44 IM7 FC
Alexa Fluor® 488 anti-mouse/human CD44 IM7 FC,ICC,IHC-P,SB
Alexa Fluor® 647 anti-mouse/human CD44 IM7 FC,ICC,SB
Pacific Blue™ anti-mouse/human CD44 IM7 FC
Alexa Fluor® 700 anti-mouse/human CD44 IM7 FC,SB
PE/Cyanine7 anti-mouse/human CD44 IM7 FC
APC/Cyanine7 anti-mouse/human CD44 IM7 FC
PerCP/Cyanine5.5 anti-mouse/human CD44 IM7 FC
PerCP anti-mouse/human CD44 IM7 FC
Brilliant Violet 421™ anti-mouse/human CD44 IM7 FC,ICC,IHC-P
Brilliant Violet 570™ anti-mouse/human CD44 IM7 FC
Brilliant Violet 785™ anti-mouse/human CD44 IM7 FC
Brilliant Violet 510™ anti-mouse/human CD44 IM7 FC
Ultra-LEAF™ Purified anti-mouse/human CD44 IM7 FC,CyTOF®,ELISA,IHC,IP,CMCD,Stim
Brilliant Violet 650™ anti-mouse/human CD44 IM7 FC
Purified anti-mouse/human CD44 (Maxpar® Ready) IM7 FC,CyTOF®
Alexa Fluor® 594 anti-mouse/human CD44 IM7 ICC,FC,IHC-P,SB
PE/Dazzle™ 594 anti-mouse/human CD44 IM7 FC
Brilliant Violet 711™ anti-mouse/human CD44 IM7 FC
APC/Fire™ 750 anti-mouse/human CD44 IM7 FC
TotalSeq™-A0073 anti-mouse/human CD44 IM7 PG
TotalSeq™-C0073 anti-mouse/human CD44 IM7 PG
TotalSeq™-B0073 anti-mouse/human CD44 IM7 PG
Spark YG™ 570 anti-mouse/human CD44 IM7 FC,IHC-P
Spark YG™ 593 anti-mouse/human CD44 IM7 FC
TotalSeq™-D0073 anti-mouse/human CD44 IM7 PG
Brilliant Violet 750™ anti-mouse/human CD44 IM7 FC
PerCP/Fire™ 806 anti-mouse/human CD44 IM7 FC
Spark Red™ 718 anti-mouse/human CD44 IM7 FC
PE/Fire™ 810 anti-mouse/human CD44 IM7 FC
Spark Blue™ 550 anti-mouse/human CD44 (Flexi-Fluor™) IM7 FC
Spark Red™ 718 anti-mouse/human CD44 (Flexi-Fluor™) IM7 FC
Go To Top Version: 9    Revision Date: 04.21.2022

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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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.

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