Anti-5-Fluorouracil (mouse IgG1, λ)

CatalogNo.NM-MA-007 data sheet

Specificity

  • 5-FU ; 5-Fluorouracil

Background

5-Fluorouracil (5-FU) is a pyrimidine analogue and inhibits an enzyme called thymidylate synthetase, which results in inhibition of DNA replication. Thus, 5-FU is used as a drug in the treatment of cancers including colorectal cancer, pancreatic cancer and skin cancer.

Clone

  • H3-17

Antigen

  • 5-Fluorouridine－BSA（5-FU-BSA）[5-FU]／[BSA]=10.2
  • Related Product : 5-FU-BSA (Cat#NM-MA-R001, 50 uL/Vial)

Form

  • This hybridoma (clone 5-FU) culture supernatant was collected and precipitated with ice-cold ammonium sulfate.After centrifugation, the pellet dissolved in small volume of double-distilled water wasdialysed against PBS.The dialysate was then frozen.
  • No preservative is contained.

Storage

  • This antibody is stored at -20 to -80℃.
  • It is stable for at least 1 year stored at or below -20℃.
  • It should be divided into small quantity to avoid freezing and thawing.

Applications

  • ELISA ; 1: 1000
  • Western blotting ; Not tested
  • Immunocytochemistry ; Not tested
  • Immunoprecipitation ; Not tested
  • Immunohistochemistry ; Not tested
  • Flow cytometry ; Not tested

Optimal dilutions/concentrations should be determined by end user.

Cross reactivity

  • Mouse

ELISA Results

ELISA Protocol

  1. 5-FU-BSA あるいは対照BSA 各々50 μg/50 μL PBS をELISA 用96 穴マイクロタイタープレートの各穴に分注し、37 ℃で一晩乾燥させてプレートに付着させます。（または4℃で一晩静置。）
  2. プレートを0.05% Tween 20 を含むPBS (PBS-T) 150μL で5 回洗浄後、2% FBS（牛胎児血清）溶液 150 μLを加え、37 ℃ で30 分間インキュベートすることにより、抗体の非特異的吸着をブロックします。
  3. PBS-T で5 回洗浄後、各濃度に希釈した抗体溶液（1 次抗体）100 μL を加え、37 ℃ で30 分間反応させます。
  4. PBS-T で5 回洗浄後、3000 倍に希釈した2 次抗体 (Peroxidase-F(ab’)2 fragment of Goat anti-mouse IgG,Zymed) 100 μL を加え、37 ℃ で30 分間反応させます。
  5. PBS-T で5 回、Citrate-phosphate buffer (pH 5.0)で1 回洗浄後、基質溶液 (o-Phenylene diamine 0.4 mg/mL, H2O2 0.06 %) 100 μL を加え、37 ℃ で正確に30 分間反応させます。
  6. 2.0 M H2SO4 50 μL で反応を停止させた後、492 nm での発色度を96 穴マイクロプレート用光度計で測定します。

Note

  • For research use only. Not for clinical diagnosis.

Related products

  • Anti-5FU プロテインGカラム精製品/Anti-5FU purified with protein G column (Cat#NM-MA-004)
  • BSA付き抗原/5-FU-BSA (Cat#NM-MA-R001)

Publication References

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