Endostatin (COL18A1) Mouse Monoclonal Antibody [Clone ID: 1837-46]

CAT#: BM4098

Endostatin (COL18A1) mouse monoclonal antibody, clone 1837-46, Purified

Specifications

Product Data

• Clone Name: 1837-46
• Applications: IHC
• Recommended Dilution: Immunohistochemistry on Frozen Sections: 5-10 µg/ml (1/40-1/80).
  Immunohistochemistry on Paraffin Sections: 5-10 µg/ml (1/40-1/80). Microwave pretreatment in citrate buffer is recommended for antigen retrieval in paraffin embedded tissues.
  Recommended Positive Control: Human tonsil.
• Reactivities: Human
• Host: Mouse
• Isotype: IgG2b
• Clonality: Monoclonal
• Immunogen: Recombinant Human Endostatin (the carboxy-terminal domain)
• Specificity: This Monoclonal antibody Clone 1937-46 is directed against the Endostatin portion (the carboxy-terminal domain) of Collagen XVIII.
• Formulation: Stock solution contains PBS, pH 7.2 with 5 mg/ml BSA as a stabilizer and 0.09% Sodium Azide as preservative
  State: Purified
  State: Lyophilized purified Ig fraction
• Reconstitution Method: Restore with 0.5 ml distilled water.
• Concentration: 0.4 mg/ml (after reconstitution)
• Purification: Affinity Chromatography
• Conjugation: Unconjugated
• Storage: Store lyophilized at 2-8°C for 6 months or at -20°C long term.
  After reconstitution store the antibody undiluted at 2-8°C for one month
  or (in aliquots) at -20°C long term.
  Avoid repeated freezing and thawing.
• Stability: Shelf life: one year from despatch.
• Gene Name: collagen type XVIII alpha 1 chain
• Database Link:
  Entrez Gene 80781 Human
  UniProt ID P39060
• Background: Endostatin is a powerful inhibitor of endothelial cell proliferation and angiogenesis and shows widespread distribution, mainly in vessel walls and some other basement membrane zones.
• Synonyms: COL18A1, Endostatin, KNO1
• Note: Protocol: Protocol for Immunohistochemistry on paraffin sections:
  1) Deparaffinize slides in chloroform for 30min and rehydrate to distilled water
  2) Antigen retrieval: microwave treatment (700W) for 15min in 300ml 0.01M citrate buffer pH 6.0.
  3) Incubate slides for 15min in 200ml methanol/4.8ml 30%H2O2
  4) Rinse slides 2x in distilled water and for 5min in TBS, pH 7.5
  5) Block with normal swine serum (NSS, Jackson Immunoresearch #014-000-121) diluted 1:10 in Tris buffered saline pH 7.5, containing 0.1% bovine serum albumin (BSA, Calbiochem). Cover sections with NSS and incubate for 15min in a humid chamber at room temperature (RT).
  6) Remove serum from sections (do not rinse)
  7) Incubate with primary antibody diluted in TBS/0.1%BSA, pH 7.5 for 60min in a humid chamber at room temperature
  8) Rinse sections in TBS, pH 7.5 for 5min, 2x
  9) Incubate with secondary antibody (rabbit anti mouse IgG+IgM-biotin, Dako E 0464) 1:400 diluted in TBS/0.1%BSA, pH 7.5 for 30min in a moist chamber, room temperature. Preincubate now StreptABComplex /HRPO in TBS/0.1%BSA, pH 7.5!
  10) Rinse sections in TBS, pH 7.5 for 5min, 2x
  11) Incubate with StreptABComplex /HRPO (DAKO, K 0377) for 30min in a humid chamber at room temperature
  12) Rinse sections in TBS, pH 7.5 for 5min, 2x
  13) DAB-staining Prepare 3,3 - diaminobenzidintetrahydrochloride (DAB) according the manufacturers instructions (e.g. DAB tablets) and incubate slides for 1-15 min. Check color reaction to avoid overstaining.
  14) Rinse sections in TBS, pH 7.5 for 5min, 2x
  15) Counterstain in haematoxylin for 1min and rinse in cold tap water for 10min
  16) Mount slides with coverslips