CD99 antigen-like 2 (CD99-L2) is a 45 kDa type I transmembrane glycoprotein in the CD99 family of molecules (1‑3). The major form of human CD99-L2 cDNA encodes a 262 amino acid (aa) precursor with a 25 aa predicted signal sequence, a 160 aa extracellular domain (ECD), a 21 aa transmembrane (TM) segment, and a 56 aa cytoplasmic region (4). This form is called the long form, or isoform E3’-E4’-E3-E4. Other isoforms include the muscle E3-E4, missing aa 45‑93, and E4, a short form missing aa 45‑116 (1, 4). E3-E4 and E3’-E3-E4 forms are the major isoforms in mouse and rat, respectively (1). Human forms that diverge at Pro 45 (145 aa precursor) or Met 180 (173 aa precursor) have been sequenced, and would be predicted to lack a TM segment (4, 5). None of the forms contain predicted N-linked glycosylation sites within the ECD, but O-linked glycosylation is likely (1, 2). The ECD of the human CD99-L2 isoform E3-E4 shares 85%, 75% and 70% aa identity with the corresponding forms of mouse, rat, and bovine CD99-L2, respectively. The human CD99 and CD99-L2 ECDs share only about 35% aa identity, but both contain three conserved acidic motifs and are thought to originate from the same ancestral gene (1, 2). The nearly ubiquitous expression of CD99-L2 is similar to that of CD99. Human CD99-L2 cDNA is detected in most organs, but not in thymus (1). In the mouse, protein is detectable in lung, thymocytes, mouse leukocytes and vascular endothelial cells (1, 3, 6). The endothelial cell CD99-L2 is reported to mediate cell aggregation and neutrophil or monocyte, but not lymphocyte, extravasation to inflamed tissue in vivo (3, 6).
Human CD99‑L2 Antibody
R&D Systems | Catalog # AF5185
Key Product Details
Species Reactivity
Validated:
Human
Cited:
Human, Transgenic Mouse
Applications
Validated:
Western Blot
Cited:
Immunohistochemistry, Immunoprecipitation
Label
Unconjugated
Antibody Source
Polyclonal Sheep IgG
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Product Specifications
Immunogen
Chinese hamster ovary cell line CHO-derived recombinant human CD99‑L2
Val20-Ala188
Accession # Q8TCZ2
Val20-Ala188
Accession # Q8TCZ2
Specificity
Detects human CD99‑L2 in direct ELISAs and Western blots.
Clonality
Polyclonal
Host
Sheep
Isotype
IgG
Scientific Data Images for Human CD99‑L2 Antibody
Detection of Human CD99‑L2 by Western Blot.
Western blot shows lysates of HUVEC human umbilical vein endothelial cells. PVDF Membrane was probed with 1 µg/mL of Sheep Anti-Human CD99-L2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5185) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). A specific band was detected for CD99-L2 at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.Detection of CD99-L2 by Western Blot
Validation of LC-MS/MS measurements with selected proteinsT(4;14) MM cell lines were treated with MMSET shRNAs for 3 days. (A) Whole cell lysates were prepared from infected cells and protein levels were analyzed by Western-blot. b-actin was used as an internal control. KMS12BM is a non-t(4;14) MM cell line, others are t(4;14) MM cell lines. (B) Determination of SLAMF7 expression in MM cell lines by indirect flow cytometric analysis. Ab control, isotype mouse IgG. ShLuc, control shRNA; Sh#4 and Sh#5, MMSET shRNAs. KMS11, KMS18, KMS28BM, NCI-H929 and OPM-2 are t(4;14) positive, other cell lines are t(4;14) negative. Results indicated SLAMF7 was universally over-expressed in t(4;14) MM cell lines and down-regulated by MMSET knockdown treatments. (C) Determination of SLAMF7 expression in primary MM cells by direct flow cytometric analysis. Ab control, mouse IgG2b-FITC. Patient samples N089, N104 and N140 are t(4;14) positive, and patient samples N048, N051 and N084 are t(4;14) negative. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/23900284), licensed under a CC-BY license. Not internally tested by R&D Systems.Applications for Human CD99‑L2 Antibody
Application
Recommended Usage
Western Blot
1 µg/mL
Sample: HUVEC human umbilical vein endothelial cells
Sample: HUVEC human umbilical vein endothelial cells
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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.
Calculators
Background: CD99-L2
References
- Suh, Y.H. et al. (2003) Gene 307:63.
- Park, S.H. et al. (2005) Gene 353:177.
- Schenkel, A.R. et al. (2007) Cell Commun. Adhes. 14:227.
- Swissprot Accession # Q8TCZ2.
- Entrez Accession # EAW99391.
- Bixel, G. et al. (2007) Blood 109:5327.
Long Name
CD99 Molecule-like 2
Alternate Names
CD99B, CD99L2, MIC2L1
Gene Symbol
CD99L2
UniProt
Additional CD99-L2 Products
Product Documents for Human CD99‑L2 Antibody
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human CD99‑L2 Antibody
For research use only
Related Research Areas
Citations for Human CD99‑L2 Antibody
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Cellular Response to Hypoxia Protocols
- R&D Systems Quality Control Western Blot Protocol
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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