Human CD97 Antibody

Catalog #: MAB25291 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB25291
MAB25291-SP

Cell Adhesion Mediated by CD97 and Neutralization by Human CD97 Antibody.

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Human CD97 Antibody Summary

Species Reactivity

Human

Specificity

Detects human CD97 in ELISAs. In sandwich immunoassays, no cross-reactivity or interference with recombinant human (rh) CD55, rhEGF, rhEMR2, or recombinant mouse CD97 is observed.

Source

Monoclonal Mouse IgG_2B Clone # 380905

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

Mouse myeloma cell line NS0-derived recombinant human CD97
Gln21-Gln398
Accession # NP_001775.2

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.

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Applications

Recommended Concentration

Sample

Human CD97 Sandwich Immunoassay

Recommended Concentration

Reagent

ELISA Capture (Matched Antibody Pair)

2-8 µg/mL

Use in combination with:

Detection Reagent: Human CD97 Biotinylated Antibody (Catalog # BAF2529)

Standard: Recombinant Human CD97 Protein, CF (Catalog # 2529-CD)

Neutralization

Measured by its ability to neutralize CD97-mediated adhesion of human red blood cells. Hamann, J. et al. (1996) J. Exp. Med. 184:1185. The Neutralization Dose (ND₅₀) is typically 0.25-1.25 µg/mL in the presence of 4 µg/mL Recombinant Human CD97.

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.

Scientific Data

Neutralization

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Cell Adhesion Mediated by CD97 and Neutralization by Human CD97 Antibody. Recombinant Human CD97 (Catalog # 2529-CD), immobil-ized onto a microplate, supports the adhesion of human red blood cells in a dose-dependent manner (orange line). Adhesion elicited by Recombinant Human CD97 (4 µg/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human CD97 Monoclonal Antibody (Catalog # MAB25291). The ND₅₀ is typically 0.25-1.25 µg/mL.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile 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.

Background: CD97

CD97 is a 95‑100 kDa member of a protein group known as the LNB-TM7 protein family that evolved from genes of the secretin receptor superfamily (1‑3). Molecules in this family are unique hybrid structures consisting of EGF-like modules coupled to class B G-protein 7-transmembrane (TM) domains by a glycosylated (mucin) stalk. Human CD97 is synthesized as an 835 amino acid (aa) precursor that contains a 20 aa signal sequence, a 532 aa extracellular domain (ECD), a 238 aa “transmembrane” region that includes seven TM segments, and a 45 aa cytoplasmic tail (4). Within the 532 aa ECD, the first 236 aa contains five EGF-like domains, the C-terminal four of which bind calcium, and a juxtamembrane 296 aa RGD-containing mucin stalk (4, 5). The stalk is both glycosylated and proteolytically cleaved (at aa 530) to create a noncovalently linked 65‑70 kDa glycosylated extracellular alpha -subunit and a 28 kDa 7-TM membrane-bound beta -subunit (4). There are two known alternate splice forms in human. Isoform # 1 contains four EGF-like domains (domain # 1, 2, 3 and 5), while isoform # 2 contains three EGF-like domains (domain # 1, 2 and 5) (1, 4, 6). The ECD in isoform 1 is 60 kDa in size, while the ECD in isoform 2 is 55 kDa in size (native molecular weight). The five EGF-like domain region in human shares 55% aa sequence identity with that in mouse. Cells known to express CD97 include monocytes, macrophages, T cells, select B cells, dendritic cells and, potentially, vascular and visceral smooth muscle cells (1, 7). There are at least two ligands for CD97. One is chrondroitin sulfate that binds only to the full-length (five domain) form of CD97. Binding is dependent on the presence of EGF-like domain # 4 (3). The second ligand for CD97 is CD55, a GPI-linked cell surface molecule with short consensus repeats that regulates complement activation on cell surfaces (1, 5, 7). CD97 EGF-like domains # 1 and 2 bind CD55 while domain # 5 stabilizes the CD97 molecule. The shortest CD97 isoform shows the strongest binding to CD55 (7, 8).

References