产品信息

• 产品组分信息

  本产品提供以下试剂或组分：

  NEB #	名称	组分货号	储存温度	数量	浓度
  M0240S     -20       Fpg M0240SVIAL -20 1 x 0.065 ml 8,000 units/ml   NEBuffer™ 1 B7001SVIAL -20 1 x 1.25 ml 10 X   Recombinant Albumin, Molecular Biology Grade B9200SVIAL -20 1 x 0.6 ml 20 mg/ml
  M0240L     -20       Fpg M0240LVIAL -20 1 x 0.32 ml 8,000 units/ml   NEBuffer™ 1 B7001SVIAL -20 1 x 1.25 ml 10 X   Recombinant Albumin, Molecular Biology Grade B9200SVIAL -20 1 x 0.6 ml 20 mg/ml

• 特性和用法

  单位定义

  1 单位指在 10 μl 反应体系中，37℃ 条件下，1 小时内能切割 10 pmol 含单个与胞嘧啶配对的 8-氧代鸟嘌呤的 34 bp 寡核苷酸双链所需要的酶量。

  反应条件

  1X NEBuffer™ 1
  Supplement with 100 µg/ml Recombinant Albumin, Molecular Biology Grade
  Incubate at 37°C

  1X NEBuffer™ 1
  10 mM Bis-Tris-Propane-HCl
  10 mM MgCl₂
  1 mM DTT
  (pH 7 @ 25°C)

  彗星实验稀释

  彗星试验的推荐稀释倍数：1:10³ 到 1:10⁴（4,5,6）。具体操作说明请登陆：http://cometassay.com。

  贮存溶液

  20 mM Tris-HCl
  50 mM NaCl
  0.5 mM EDTA
  200 µg/ml BSA
  50% Glycerol
  pH 8 @ 25°C

  热失活

  60°C for 10 minutes

  单位活性检测条件

  10 μl 总反应体系，含 1X NEBuffer 1、10 pmol 荧光标记寡核苷酸双链和 100 µg/ml BSA。

• 相关产品

  单独销售的组分

  • NEBuffer 1
  • 重组白蛋白, 分子生物学级（不含动物成分）

• 参考文献

  1. Tchou, J. et al. (1994). Substrate specificity of Fpg protein. J. Biol.Chem.. 269, 15318-15324.
  2. Hatahet, Z. et al. (1994). Newsubstrates for old enzymes. J. Biol.Chem.. 269, 18814-18820.
  3. Boiteux, S., O’Connor, T. and Laval, J. (1987). Formamidopyrimidine-DNA glycosylase of Escherichia coli: cloning and sequencing of the fpg structural gene and overproduction of the protein. EMBOJ.. 5, 18814-18820.
  4. Singh, N., McCoy, M., Tice, R. and Schneider, L. (1988). A simple technique for quantitation of low levels of DNA damage in individual cells. Exp.Cell Res.. 175, 184-191.
  5. Collins, A., Duthie, S. and Dobson, V. (1993). Direct enzymatic detection of endogenous oxidative base damage in human lymphocyte DNA. Carcinogenesis. 14, 1733-1735.
  6. Collins, A., Dusinska, M., Gedik, C. and Stetina, R. (1996). Oxidative damage to DNA: do we have a reliable biomarker?. Environ.Health Perspect.. 104, 465-469.
  7. Pflaum, M., Will, O., Mahler, H.-C. and Epe, B. (1998). DNA oxidation products determined with repair endonucleases in mammalian cells: types, basal levels and influence of cell proliferation. FreeRad. Res.. 29, 585-594.
  8. Hartwig, A., Dally, H. and Schlepegrell, R. (1996). Sensitive analysis of oxidative DNA damage in mammalian cells: use of the bacterial Fpg protein in combination with alkaline unwinding. Toxicol.Lett.. 88, 85-90.
  9. Czene, S. and Harms-Ringdahl, M. (1995). Detection of single strand breaks and formamidopyrimidine-DNA glycosylase-sensitive sites in DNA of cultured human fibroblasts. Mutat.Res.. 336, 235-242.

操作说明、说明书 & 用法

• 操作说明

  1. Comet Assay – Modified for Detection of Oxidized Bases Using the Repair Endonucleases Fpg, hOGG1 and Endonuclease III (Nth)

• 使用指南

  • DNA Damage and PreCR

工具 & 资源

• 选择指南

  • Activities of DNA Repair Enzymes and Structure-specific Endonucleases
  • DNA Repair Enzymes on Damaged and Non-standard Bases
  • Properties of DNA Repair Enzymes and Structure-specific Endonucleases

FAQs & 问题解决指南

• FAQs

  1. What is the activity of Fpg in the NEBuffers 1-4?
  2. Does Fpg cleave hydroxymethyl uracil?
  3. Does Fpg cut only one strand, or do they cause a double-strand break?
  4. What is the Molecular Weight of Fpg?
  5. What is the difference between using Fpg and hOOG1?
  6. Does the Fpg contain a tag?
  7. What buffer will work with both Endonuclease III and Fpg?
  8. What substrate is Fpg tested on?
  9. What is the activity of Fpg in rCutSmart Buffer?