Recombinant Mouse CD99-Fc Chimera (carrier-free)

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Regulatory Status
RUO
Other Names
CD99 antigen, Paired immunoglobin-like type 2 receptor-ligand, PILR-L
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CD99_Mouse_RECOM_BA_1_030423
Immobilized recombinant mouse CD99-Fc Chimera at 5 µg/mL binds to His tagged-recombinant mouse PILRα in a dose dependent manner. HRP anti-His Tag (Cat. No. 652504) was used to detect the binding. The ED50 for this effect is 0.3 – 6.0 µg/mL.
  • CD99_Mouse_RECOM_BA_1_030423
    Immobilized recombinant mouse CD99-Fc Chimera at 5 µg/mL binds to His tagged-recombinant mouse PILRα in a dose dependent manner. HRP anti-His Tag (Cat. No. 652504) was used to detect the binding. The ED50 for this effect is 0.3 – 6.0 µg/mL.
  • CD99_Mouse_RECOM_BA_2_030423
    Stability testing for recombinant mouse CD99-Fc Chimera. Recombinant mouse CD99-Fc Chimera was aliquoted in PBS at 0.2 mg/mL and one aliquot was kept at 4°C (Control), and another was frozen and thawed four times (4x Freeze/Thaw). The samples were tested by their ability to bind to His tagged-recombinant mouse PILRα in a dose dependent manner. HRP anti-His Tag (Cat. No. 652504) was used to detect the binding. The ED50 for this effect is 0.3 – 6.0 µg/mL.
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798904 25 µg 195 CHF
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798906 100 µg 790 CHF
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Description

Mouse CD99 (mCD99) is a highly O-glycosylated transmembrane protein that is expressed in endothelial cells and various immune cells, including thymocytes, lymphocytes, monocytes, and neutrophils. CD99 is encoded by the pseudoautosomal gene MIC2. This gene is found in the pseudoautosomal region 1 (PAR1) at the end of the short arm of chromosomes X and Y. The mouse CD99 contains 175 amino acid (aa) with a 28 aa signal sequence, a 109 aa extracellular domain (aa 29-137), a 21 aa transmembrane segment (aa 138-158), and a 17 aa cytoplasmic region (aa 159-175). Mouse CD99 shares 45% and 50% aa sequence identity with the human and rat proteins, respectively. As a cellular ligand for the paired immunoglobulin-like type 2 receptors (PILRs), mCD99 is also called paired immunoglobin-like type 2 receptor-ligand (PILR-L). Mouse CD99 binding to mPILRα relies on the recognition of the Sialylated O-linked sugar chains of mouse CD99, widely involved in the regulation of immune responses. In addition, CD99 regulates various crucial biological processes, including cell adhesion, apoptosis, cell differentiation, and especially plays an important role in the transendothelial migration (TEM) of leukocytes, the critical event for inflammation. CD99 not only serves as a robust marker in cancer, but also as a potential therapeutic target for the treatment of cancers. CD99 influences processes and malignancy associated with several tumors. CD99 is frequently overexpressed in Ewing sarcoma (EWS) and specific subtypes of leukemia. The blockade and knockdown of CD99 in EWS tumor cells promote the inhibition of EWS tumor growth and metastasis. CD99 also modulates the expression of heat shock protein 70 (HSP70), thereby promoting natural killer (NK)-cell activity against acute lymphoblastic leukemia cells. In contrast, CD99-forced expression showed a dramatic negative impact on the survival of osteosarcoma cells, likely through the regulation of caveolin-1 and c-Src kinase activity.

Product Details
Technical Data Sheet (pdf)

Product Details

Source
Recombinant mouse CD99-Fc Chimera, amino acid sequence containing Ala27-Gly137 (Accession No. Q8VCN6.1) with a linker (GSSR), and a C-terminal hIgG1-Fc (Pro100-Lys330) was expressed in CHO cells.
Molecular Mass
The 346 amino acid recombinant protein has a predicted molecular mass of approximately 36.88 kD. The DTT-reduced and non-reduced protein migrates at approximately 50 kD and 90 kD by SDS-PAGE, respectively. The predicted N-terminal amino acid is Ala.
Purity
> 95%, as determined by Coomassie stained SDS-PAGE.
Formulation
0.22 µm filtered protein solution is in PBS
Endotoxin Level
Less than 0.1 EU per µg cytokine as determined by the LAL method.
Concentration
25 µg size is bottled at 200 µg/mL. 100 µg size and larger sizes are lot-specific and bottled at the concentration indicated on the vial. To obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.
Storage & Handling
Unopened vial can be stored between 2°C and 8°C for up to 2 weeks, at -20°C for up to six months, or at -70°C or colder until the expiration date. For maximum results, quick spin vial prior to opening. The protein can be aliquoted and stored at -20°C or colder. Stock solutions can also be prepared at 50 - 100 µg/mL in appropriate sterile buffer, carrier protein such as 0.2 - 1% BSA or HSA can be added when preparing the stock solution. Aliquots can be stored between 2°C and 8°C for up to one week and stored at -20°C or colder for up to 3 months. Avoid repeated freeze/thaw cycles.
Activity
Immobilized recombinant mouse CD99-Fc Chimera at 5 µg/mL binds to His tagged-recombinant mouse PILRα in a dose dependent manner. HRP anti-His Tag (Cat. No. 652504) was used to detect the binding. The ED50 for this effect is 0.3 – 6.0 µg/mL.
Application

Bioassay

Application Notes

BioLegend carrier-free recombinant proteins provided in liquid format are shipped on blue-ice. Our comparison testing data indicates that when handled and stored as recommended, the liquid format has equal or better stability and shelf-life compared to commercially available lyophilized proteins after reconstitution. Our liquid proteins are validated in-house to maintain activity after shipping on blue ice and are backed by our 100% satisfaction guarantee. If you have any concerns, contact us at tech@biolegend.com.

Antigen Details

Structure
Homodimer
Distribution

Expressed on leukocytes and endothelial cell contacts. Detected in a wide range of T-cells, including CD4-/CD8- and CD4+/CD8+ thymocytes 

Function
T-cell adhesion, participates in T-cell recruitment into inflamed skin, and leukocyte extravasation. Acts at the same site as, but independently of, PECAM1
Interaction
Annexin II, E2H, Ferritin heavy chain 1, karyopherin beta 1, and TRIP13
Ligand/Receptor
Paired immunoglobulin like receptors (PILRs)
Bioactivity
Measured by its ability to bind recombinant mouse PILRα in functional ELISA
Cell Type
B cells, Basophils, Dendritic cells, Fibroblasts, Macrophages, Mast cells, NK cells, Platelets, T cells
Biology Area
Immunology
Molecular Family
CD Molecules
Antigen References
  1. Goodfellow PN, et al. 1988. Philos Trans R Soc Lond B Biol Sci. 322:145-54.
  2. Husak Z, et al. 2012. Cell Death Dis. 3:e425.
  3. Olivia L, et al. 2007. J Immunol. 178:1136-1143.
  4. Manara MC, et al. 2006. Mol Biol Cell. 17:1910-21.
  5. Park SH, et al. 2005. Gene. 353:177-188.
  6. Pasello M, et al. 2018. J Cell Commun Signal. 12:55-68.
  7. Rocchi A, et al. 2010. J Clin Invest. 120:668-680.
  8. Schenkel AR, et al. 2002. Nat Immunol. 3:143-150.
  9. Scotlandi K, et al. 2006. Eur J Cancer. 42:91-6.
  10. Watson RL, et al. 2015. J Exp Med. 212:1021-1041.
Gene ID
673094 View all products for this Gene ID
UniProt
View information about CD99 on UniProt.org

Related FAQs

Why choose BioLegend recombinant proteins?

     • Each lot of product is quality-tested for bioactivity as indicated on the data sheet.
     • Greater than 95% Purity or higher, tested on every lot of product.
     • 100% Satisfaction Guarantee for quality performance, stability, and consistency.
     • Ready-to-use liquid format saves time and reduces challenges associated with reconstitution.
     • Bulk and customization available. Contact us.
     • Learn more about our Recombinant Proteins.

How does the activity of your recombinant proteins compare to competitors?

We quality control each and every lot of recombinant protein. Not only do we check its bioactivity, but we also compare it against other commercially available recombinant proteins. We make sure each recombinant protein’s activity is at least as good as or better than the competition’s. In order to provide you with the best possible product, we ensure that our testing process is rigorous and thorough. If you’re curious and eager to make the switch to BioLegend recombinants, contact your sales representative today!

What is the specific activity or ED50 of my recombinant protein?

The specific activity range of the protein is indicated on the product datasheets. Because the exact activity values on a per unit basis can largely fluctuate depending on a number of factors, including the nature of the assay, cell density, age of cells/passage number, culture media used, and end user technique, the specific activity is best defined as a range and we guarantee the specific activity of all our lots will be within the range indicated on the datasheet. Please note this only applies to recombinants labeled for use in bioassays. ELISA standard recombinant proteins are not recommended for bioassay usage as they are not tested for these applications.

Have your recombinants been tested for stability?

Our testing shows that the recombinant proteins are able to withstand room temperature for a week without losing activity. In addition the recombinant proteins were also found to withstand four cycles of freeze and thaw without losing activity.

Does specific activity of a recombinant protein vary between lots?

Specific activity will vary for each lot and for the type of experiment that is done to validate it, but all passed lots will have activity within the established ED50 range for the product and we guarantee that our products will have lot-to-lot consistency. Please conduct an experiment-specific validation to find the optimal ED50 for your system.

How do you convert activity as an ED50 in ng/ml to a specific activity in Units/mg?

Use formula Specific activity (Units/mg) = 10^6/ ED50 (ng/mL)

Go To Top Version: 1    Revision Date: 03.03.2023

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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