ケーススタディ 4: アプリケーション (XP®モノクローナル抗体)
#4060 Phospho-Akt (Ser473) (D9E) Rabbit mAb
XMT®は、従来のハイブリドーマ技術では成し得なかった様々なB細胞 (抗体産生細胞) の開発を可能にし、より包括的なスクリーニングを可能にしました。CSTの厳格な品質管理とXMT®を組み合わせることにより、様々なアプリケーションで優れたパフォーマンスを発揮するXP®モノクローナル抗体の選別が可能になりました。#4060 Phospho-Akt (Ser473) (D9E) XP® Rabbit mAbを、従来の方法では不可能だったすべてのアプリケーションで優れたパフォーマンスを発揮する一例としてご紹介します (Table 1 参照)。
Figure 1. Western blot analysis of extracts from PC-3 cells, untreated or LY294002/wortmannin-treated, and NIH/3T3 cells, serumstarved or PDGF-treated, using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (upper) or Akt (pan) (C67E7) Rabbit mAb #4691 (lower).
Figure 2A. Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060.
Figure 2B. Immunohistochemical analysis of paraffin-embedded PTEN heterozygous mutant mouse endometrium using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060. (Tissue section courtesy of Dr. Sabina Signoretti, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA.).
Figure 2C. Immunohistochemical analysis using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060 on SignalSlide® Phospho-Akt (Ser473) IHC Controls #8101 (paraffin-embedded LNCaP cells, untreated (left) or LY294002-treated (right)).
Figure 3. Confocal immunofluorescent analysis of C2C12 cells, LY294002-treated (left) or insulin-treated (right), using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060 (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Figure 4. Flow cytometric analysis of Jurkat cells, untreated (green) or treated with LY294002, wortmannin and U0126 (blue), using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060 compared to a nonspecific negative control antibody (red).
| No. | Antibody | Reactivity | WB | IP | IHC | Flow | IF |
|---|---|---|---|---|---|---|---|
| 4060 | Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb | H, M, R, Hm, Mk, Dm, Z, B, (C, X, Dg, Pg) | ++++ | ++++ | ++++ | ++++ | ++++ |
| 4058 | Phospho-Akt (Ser473) (193H12) Rabbit mAb | H, M, R | +++ | +++ | - | +++ | +++ |
| 3787 | Phospho-Akt (Ser473) (736E11) Rabbit mAb | H, M, (R) | - | - | ++ | - | - |
| 9271 | Phospho-Akt (Ser473) Antibody | H, M, R, Hm, Dm, B, Dg, Pg, (Mk, C, X, Hr) | +++ | ++ | - | ++ | ++ |
| 4051 | Phospho-Akt (Ser473) (587F11) Mouse mAb | H, M, R, Hm, (Mk) | +++ | +++ | - | - | - |
Table 1. Summary of Phospho-Akt (Ser473) Antibodies available from CST. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Applications Key: WB=Western Blot IP=Immunoprecipitation IHC=Immunohistochemistry Flow=Flow Cytometry IF=Immunofluorescence
Testing Data Key: (++++)=Very Highly Recommended (+++)=Highly Recommended (++)=Recommended (-)=Not Recommended N/T=Not Tested
Reactivity Key: H=Human M=Mouse R=Rat Hm=Hamster Mk=Monkey C=Chicken Dm=D.melanogaster X=Xenopus Z=Zebrafish B=Bovine Dg=Dog Pg=Pig Hr=Horse
Species enclosed in parentheses are predicted to react based on 100% sequence homology.