DIG Luminescent Detection Kit 地高辛发光检测试剂盒
英文名称： DIG Luminescent Detection Kit
型号:null    产品货号： 11363514910
价格:请致电：010-57128832,18610462672
品牌: roche

DIG Luminescent Detection Kit 地高辛发光检测试剂盒，CSPD化学发光法检测, 灵敏度可达0.03pg同源DNA; 可检测50张100cm2杂交膜。

Application

Highly sensitive and specific detection of DIG-labeled nucleic acids in Southern and northern blots and plaque or colony lifts with anti-DIG-AP conjugate and the chemiluminescence substrate CSPD.
Chemiluminescent detection with CSPD (following the instructions of the kit) is as sensitive as radioactive methods, but requires much shorter exposure times.

Product Description

Sensitivity: 0.03 pg of homologous DNA or 0.1 pg of homologous RNA is detected in a Southern or dot blot after <30-min exposure time. Single-copy genes are detected in 0.3 µg mammalian genomic DNA.



Figure 1: Detection of DIG-labeled nucleic acids with CSPD. Human genomic DNA was digested with Eco RI, separated on a 1 agarose gel, and blotted onto positively charged nylon membranes. The blots were hybridized with 50 ng/ml DIG-labeled β-actin RNA. Chemiluminescent detection was performed according to the standard DIG chemiluminescent-detection procedure, using CSPD at a 0.25-mM final concentration.

Background Information

Enzymatic dephosphorylation of the dioxetane CSPD by alkaline phosphatase leads to the metastable phenolate anion, which decomposes and emits light at 477 nm. The light emission increases with time until a constant intensity is attained.

Contents

  1. DIG-labeled Control DNA, 50 µl, [5 µg/ml]
  2. DNA Dilution Buffer, 1 ml
  3. Anti-digoxigenin-AP, Fab fragments, 100 µl, [750 U/ml]
  4. Blocking Reagent, 2 x 50 g
  5. CSPD, 1 ml, [11.6 mg/ml]

Principle

  • DIG-labeled nucleic-acid probes are used for hybridization in various blot applications.
  • After hybridization, the blots are submitted to immunological detection using a highly specific anti-digoxigenin-antibody, conjugated to alkaline phosphatase.
  • The addition of the chemiluminescence substrate CSPD leads to a distinct luminescent signal that is recorded on X-ray or Polaroid film.
  • Exposure time varies between 5 and 20 min, and because of the long-lasting glow, multiple exposures can be taken over several days.
  • Blots developed with CSPD can be easily stripped and reprobed.


Figure 2: CSPD reaction.

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