Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Western Blot, Neutralization, Flow Cytometry, CyTOF-ready

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2A Clone # 380903
View all CD97 Primary Antibodies »

Product Specifications

Immunogen

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

Specificity

Detects human CD97 in direct ELISAs and Western blots. In direct ELISAs and Western blots, no cross‑reactivity with recombinant mouse CD97 is observed.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2A

Endotoxin Level

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

Scientific Data Images for Human CD97 Antibody

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

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 # MAB2529). The ND50 is typically 0.5-2.5 µg/mL.
Detection of CD97 antibody in Human Whole Blood antibody by Flow Cytometry.

Detection of CD97 in Human Whole Blood by Flow Cytometry.

Human whole blood was stained with Mouse Anti-Human CD97 Monoclonal Antibody (Catalog # MAB2529, filled histogram) or isotype control antibody (Catalog # MAB0031, open histogram), followed by Phy-coerythrin-conjugated Anti-Mouse IgG F(ab')2Secondary Antibody (Catalog # F0102B).
Detection of CD97 by Western Blot

Detection of CD97 by Western Blot

Immunoblot image are shown (A), along with densitometric analysis of (B) PDGFR beta, (C) Cyr61, (D) CD97, (E) Glypican-1, (F) MUC18/CD146 and (G) uPAR. (H) A summary of proteins detected by mass spectrometry and immunoblot. Graphs represent mean ± SEM of n = 3 independent biological experiments, shown as 1,2,3. Unpaired t-test was used as statistical test. *P < 0.05, **P < 0.01, ***P < 0.001 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36171564), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of CD97 by Western Blot

Detection of CD97 by Western Blot

Immunoblot image are shown (A), along with densitometric analysis of (B) PDGFR beta, (C) Cyr61, (D) CD97, (E) Glypican-1, (F) MUC18/CD146 and (G) uPAR. (H) A summary of proteins detected by mass spectrometry and immunoblot. Graphs represent mean ± SEM of n = 3 independent biological experiments, shown as 1,2,3. Unpaired t-test was used as statistical test. *P < 0.05, **P < 0.01, ***P < 0.001 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36171564), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human CD97 Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

2.5 µg/106 cells
Sample: Human whole blood

Western Blot

1 µg/mL
Sample: Recombinant Human CD97 (Catalog # 2529-CD) under non-reducing conditions only

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 0.5-2.5 µg/mL in the presence of 4 µg/mL Recombinant Human CD97.

Reviewed Applications

Read 1 review rated 5 using MAB2529 in the following applications:

Flow Cytometry Panel Builder

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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 is 55% aa identical to 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

  1. McKnight, A.J. and S. Gordon (1998) J. Leukoc. Biol. 63:271.
  2. Stacey, M. et al. (2000) Trends Biochem. Sci. 25:284. 
  3. Stacey, M. et al. (2003) Blood 102:2916. 
  4. Gray, J.X. et al. (1996) J. Immunol. 157:5438. 
  5. Lin, H-H. et al. (2001) J. Biol. Chem. 276:24160.
  6. Hamann, J. et al. (1995) J. Immunol. 155:1942.
  7. Jaspars, L.H. et al. (2001) Tissue Antigens 57:325.
  8. Hamann, J. et al. (1996) J. Exp. Med. 184:1185.

Alternate Names

CD97

Entrez Gene IDs

976 (Human); 26364 (Mouse)

Gene Symbol

ADGRE5

UniProt

NP_001775.2

Additional CD97 Products

Product Documents for Human CD97 Antibody

Certificate of Analysis

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Product Specific Notices for Human CD97 Antibody

For research use only

Citations for Human CD97 Antibody

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  • Human CD97 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Peripheral blood lymphocytes (PBL)
    Species: Human
    Verified Customer | Posted 05/18/2022
    Human CD97 Antibody MAB2529

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