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品質管理 - フローサイトメトリー (F)



  • 最適希釈率を決定するために、様々な希釈率で試験
  • アイソタイプコントロールとの比較により、一次抗体の非特異結合を判断
  • 既知のポジティブ/ネガティブ細胞株を用いて特異性を確認
  • パスウェイ特異的な阻害剤や活性化剤処理により、特異性を確認
  • ブロッキングペプチド、siRNA、発現ベクターを用い、染色の特異性を確認
  • フォスファターゼ処理を行い、抗体のリン酸化特異性を確認
  • ロット間の比較試験を行い再現性を確保
  • 推奨プロトコールと最適条件を決定


Titration of BrdU (Bu20a) Mouse mAb #5292.

BrdU (Bu20a) Mouse mAb #5292: Flow cytometric analysis of Jurkat cells, unincorporated (red) or after 30 minutes of BrdU incorporation (blue), using serial dilutions of #5292. The fold-induction ratio is shown in green. Optimal concentration of #5292 was determined to be 0.044 µg/ml.

Isotype Control

Isotype Control Example for #3900.

Rabbit (DA1E) mAb IgG XP® Isotype Control #3900: Flow cytometric analysis of SH-SY5Y cells using CREB (D76D11) Rabbit mAb Antibody #4820 (blue) compared to concentration matched #3900 (red).

Activation-state Specificity

Activation-state Specificity Example for #4324.

Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) #4324: Flow cytometric analysis of human whole blood, untreated (red) or treated with Human Granulocyte Colony Stimulating Factor (hG-CSF) #8930 (blue), using #4324.

Inhibitor Treatment

Inhibitor Treatment Example for #4344.

Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) #4344: Flow cytometric analysis of Jurkat cells treated with U0126 #9903 (blue) or TPA #4174 (green), using #4344.

Positive and Negative Cell Lines

Positive and Negative Cell Lines Examples for #2707.

Zap-70 (136F12) Rabbit mAb (Alexa Fluor® 647 Conjugate) #2707: Flow cytometric analysis (A) of Ramos B (blue) and Jurkat T (green) cells using #2707. Two-color flow cytometric analysis (B) of a mixed population of T and B cells (Jurkat and Ramos, respectively) using Zap-70 Antibody and a CD3 antibody. CD3-negative B cells have little or no Zap-70 staining, while CD3-positive T cells stain brightly for Zap-70 protein.

品質管理 - フローサイトメトリー (F)