Human CD99 Antibody

Catalog # Availability Size / Price Qty
AF3968
AF3968-SP
Detection of CD99 in Human CD3+ Lymphocytes by Flow Cytometry.
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Product Details
Citations (2)
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Human CD99 Antibody Summary

Species Reactivity
Human
Specificity
Detects human CD99 in direct ELISAs and Western blots. In direct ELISAs, less than 5% cross‑reactivity with recombinant mouse CD99 is observed.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant human CD99
Asp23-Asp122
Accession # P14209
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.1 µg/mL
Recombinant Human CD99 Fc Chimera (Catalog # 3968-CD)
Flow Cytometry
0.25 µg/106 cells
See below
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 
Immunocytochemistry
5-15 µg/mL
See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Data Examples

Flow Cytometry Detection of CD99 in Human CD3+ Lymphocytes by Flow Cytometry. View Larger

Detection of CD99 in Human CD3+ Lymphocytes by Flow Cytometry. Human whole blood CD3+ lymphocytes were stained with Goat Anti-Human CD99 Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF3968, filled histogram) or control antibody (Catalog # AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107).

Immunocytochemistry CD99  in HepG2 Human Cell Line. View Larger

CD99 in HepG2 Human Cell Line. CD99 was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line using Goat Anti-Human CD99 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3968) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cell membranes. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: CD99

CD99 (also named MIC2, E2 and thymic leukemia antigen) is the founding member of the CD99 family of molecules. The CD99 family contains four members; CD99, CD99L2, XG and the pseudogene CD99L1 (1, 2, 3). Native human CD99 is 32 kDa in size and exists as a type I transmembrane glycoprotein. This is referred to as the long, or type I isoform. It is synthesized as a 185 amino acid (aa) precursor that contains a 22 aa signal sequence, a 100 aa extracellular domain (ECD), a 25 aa transmembrane segment, and a 38 aa cytoplasmic region (4). The ECD contains no identifiable motifs, N‑linked glycosylation sites, or cysteine residues; it does possess sites for O-linked glycosylation. The cytoplasmic region, albeit short, does have signal transduction capability (5). There are apparently multiple isoforms for human CD99. One shows a 16 aa deletion in the ECD (aa 34‑49), a second shows a 38 aa deletion in the cytoplasmic region (aa 122‑159), and a third exhibits a three aa truncation at the C-terminus (6, 7, 8). The best studied isoform shows an Asp‑Gly substitution for the C‑terminal 27 amino acids. This is referred to as the 28 kDa type II isoform (9). The type I and II isoforms have distinctive signal transduction pathways (FAK-src for type I; PI3K plus src-ERK1/2 for type II), and mediate clearly different biological outcomes (5, 9, 10). The two numbered isoforms may or may not co‑exist on the same cells. Peripheral T cells have only the long isoform, while double-positive thymocytes express both isotypes. What is unclear is the monomeric vs. dimeric status of CD99. In mouse, CD99 reportedly forms disulfide-linked homodimers (11). In human, however, CD99 is reportedly monomeric if only a type I isoform, and a covalent heterodimer if co‑expressing type I and II isoforms (12, 13). Cells known to express CD99 include fibroblasts, neutrophils, T cells, double-positive thymocytes, CD34+ stem cells, monocytes and endothelial cells (2, 12, 14, 15). Homophilic interaction between CD99 on the neutrophil and CD99 on the endothelial cell regulates the transendothelial migration of neutrophils during inflammation (16). Human CD99 is only 48% aa identical to mouse CD99 (17).

References
  1. Wilson, M.D. et al. (2006) Physiol Genomics 27:201. 
  2. Petri, B. and M.G. Bixel (2006) FEBS J. 273:4399. 
  3. Suh, Y.H. et al. (2003) Gene 307:63. 
  4. Gelin, C. et al. (1989) EMBO J. 8:3253. 
  5. Byun, H-J. et al. (2006) J. Biol. Chem. 281:34833. 
  6. GenBank Accession # EAW98698. 
  7. GenBank Accession # EAW98699. 
  8. GenBank Accession # EAW98700. 
  9. Hahn, H-J. et al. (1997) J. Immunol. 159:2250.
  10. Scotlandi, K. et al. (2007) Oncogene Apr 30; [Epub ahead of print].
  11. Park, S.H. et al. (2005) Gene 353:177.
  12. Schenkel, A.R. et al. (2002) Nat. Immunol. 3:143.
  13. Alberti, I. et al. (2002) FASEB J. 16:1946.
  14. Imbert, A-M. et al. (2006) Blood 108:2578.
  15. Dworzak, M.N. et al. (1994) Blood 83:415.
  16. Lou, O. et al. (2007) J. Immunol. 178:1136
  17. Shiratori, I. et al. (2004) J. Exp. Med. 199:525.
Entrez Gene IDs
4267 (Human); 673094 (Mouse); 652929 (Rat)
Alternate Names
12E7; antigen identified by monoclonal antibodies 12E7, F21 and O13; CD99 antigenY homolog; CD99 molecule; CD99; E2 antigen; HBA71; MIC2 (monoclonal 12E7); MIC2; MIC2Y; MSK5X; pilr-1; PILR-L; Protein MIC2; surface antigen MIC2; T-cell surface glycoprotein E2

Product Datasheets

Citations for Human CD99 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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  1. Ectodomain shedding of CD99 within highly conserved regions is mediated by the metalloprotease meprin ? and promotes transendothelial cell migration
    Authors: Tillmann Bedau
    FASEB J, 2016;0(0):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  2. Evolutionarily conserved paired immunoglobulin-like receptor alpha (PILRalpha) domain mediates its interaction with diverse sialylated ligands.
    Authors: Sun Y, Senger K, Baginski TK, Mazloom A, Chinn Y, Pantua H, Hamidzadeh K, Ramani SR, Luis E, Tom I, Sebrell A, Quinones G, Ma Y, Mukhyala K, Sai T, Ding J, Haley B, Shadnia H, Kapadia SB, Gonzalez LC, Hass PE, Zarrin AA
    J. Biol. Chem., 2012;287(19):15837-50.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry

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