Analysis Note

ControlADAM 10 is ubiquitously expressed. MDA-MB231 (Human breast cancer cell line known to express high levels of ADAM 10).

Application

Western blot:
1:500 - 1:2,000 dilution. Jurkat whole cell lysate can be used as a positive control and an 85 kDa band can be detected which may represent precursor. A 60 kDa faint band was detected in some cell lines including Jurkat, which appears to be the processed mature protein. Overloading of the primary antibody can lead to multiple bands appearing in some preparations. The presence of glycosylated forms and breakdown products can lead to other bands. A prominent proform at near 80-85 is usually seen, with bands at 60, and below 50 kDa in most cases. The immunogen peptide is available (Cat. No. AG533) for performing antibody blocking studies.

Optimal working dilutions must be determined by end user.

Immunohistochemistry:
A previous lot of this antibody was used to detect ADAM10 in formalin-fixed, paraffin-embedded renal tissue and renal cell carcinoma tissue (Gutwein, P., et al. EUROPEAN JOURNAL OF CANCER. 45(2009):478–489).

Immunofluorescence:
A previous lot of this antibody was used to detect ADAM10 in RCC4 (renal cell carcinoma) and A498 cells (Gutwein, P., et al. EUROPEAN JOURNAL OF CANCER. 45(2009):478–489).

This Anti-ADAM 10 Antibody, C-terminus is validated for use in WB for the detection of ADAM 10.

General description

ADAM10 is a member of the ADAM (a disintegrin and metalloprotease like domain) family. It cleaves the membrane-bound precursor of TNF-alpha at 76-Ala-|-Val-77 to its mature soluble form. It is responsible for the proteolytic release of several cell-surface proteins, including heparin binding epidermal growth-like factor, ephrin-A2 and for constitutive and regulated alpha-secretase cleavage of amyloid precursor protein (APP). It contributes to the normal cleavage of the cellular prion protein and is involved in the cleavage of the adhesion molecule L1 at the cell surface and in released membrane vesicules, suggesting a vesicle-based protease activity.Tumor-necrosis factor alpha is a proinflammatory cytokine and contributes to a variety of inflammatory disease responses and programmed cell death. Notch receptor and its ligand participate in cell fate decisions during vertebrate development and are associated with several human disorders, including a T-cell lymphoma. TNA-alpha, notch and its ligand delta are all membrane-bond molecules, which are cleaved by proteases to release mature proteins or functional receptor. ADAM10-, a metalloprotease-disintegrin in the family of mammalian ADAM (for a disintegrin and metalloproteinase domain), was recently identified to cleave TNF-alpha, notch and its ligand delta (1-3). The genes encoding human, mouse, and bovine ADAM10 were recently cloned and designated ADAM10, kuzbanian (KUZ), and MADM, respectively, (1,2,4). ADAM10mRNA is expressed in a variety of human and bovine tissues.

Immunogen

Epitope: a.a. 732-748 of human ADAM 10

Raised against a peptide corresponding to amino acids 732-748 of human ADAM 10 (Rosendahl, M.S., 1997). This sequence is identical to those of bovine and rat origins and differs from that of mouse ADAM 10 by one amino acid (Pan, D., 1997; Howard, L., 1996).

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Linkage

Replaces: AB19031

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Format: Purified

Quality

Routinely evaluated by Western Blot on RAW264.7 lysates.

Western Blot: 1:500 dilution of this lot detected ADAM 10 on 10 µg of RAW264.7 lysates

Specificity

Recognizes ADAM 10, C-terminus.

Storage and Stability

Stable for 1 year at -20°C in undiluted aliquots from date of receipt.

Handling Recommendations:
Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Target description

80-85 kDa