• Western blot analysis of extracts from PC-3 cells, untreated or LY294002/wortmannin-treated, and NIH/3T3 cells, serum-starved or PDGF-treated, using Phospho-Akt (Ser473) (D9E) XP ® Rabbit mAb (upper) or Akt (pan) (C67E7) Rabbit mAb #4691 (lower). • Immunohistochemical analysis of paraffin-embedded MDA-MB-468 xenograft using Phospho-Akt (Ser473) (D9E) XP ® Rabbit mAb (left) or PTEN (138G6) Rabbit mAb #9559 (right).

Note the presence of P-Akt staining in the PTEN deficient MDA-MB-468 cells. • Immunohistochemical analysis of paraffin-embedded human breast carcinoma comparing SignalStain ® Antibody Diluent #8112 (left) to TBST/5% normal goat serum (right) using Phospho-Akt (Ser473) (D9E) XP ® Rabbit mAb #4060. • Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-Akt (Ser473) (D9E) XP ® Rabbit mAb. • Immunohistochemical analysis using Phospho-Akt (Ser473) (D9E) XP ® Rabbit mAb on SignalSlide® Phospho-Akt (Ser473) IHC Controls #8101 (paraffin-embedded LNCaP cells, untreated (left) or LY294002-treated (right)). • Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-Akt (Ser473) (D9E) XP ® Rabbit mAb. • Immunohistochemical analysis of paraffin-embedded PTEN heterozygous mutant mouse endometrium using Phospho-Akt (Ser473) (D9E) XP ® Rabbit mAb. (Tissue section courtesy of Dr.

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Sabina Signoretti, Brigham and Women's Hospital, Harvard Medical School, Boston, MA.) • Immunohistochemical analysis of paraffin-embedded U-87MG xenograft, untreated (left) or lambda phosphatase-treated (right), using Phospho-Akt (Ser473) (D9E) XP ® Rabbit mAb. • Confocal immunofluorescent analysis of C2C12 cells, LY294002-treated (left) or insulin-treated (right), using Phospho-Akt (Ser473) (D9E) XP ® Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor ® 555 phalloidin #8953 (red).

Jan 25, 2008  Akt is a protein serine/threonine kinase that is involved in the regulation of diverse cellular processes. Phosphorylation of Akt at regulatory residues Thr-308 and Ser-473 leads to its full activation. The protein phosphatase 2A (PP2A) has long been known to negatively regulate Akt activity. The PI3K-PKB/Akt pathway is highly conserved, and its activation is tightly controlled via a multistep process (as shown in Fig. 1) Activated receptors directly stimulate class 1A PI3Ks bound via their regulatory subunit or adapter molecules such as the insulin receptor substrate (IRS) proteins. Instrukciya po razvedeniyu hloramina.

Blue pseudocolor = DRAQ5 ®#4084 (fluorescent DNA dye). • Flow cytometric analysis of Jurkat cells, untreated (green) or treated with LY294002 #9901, wortmannin #9951 and U0126 #9903 (blue), using Phospho-Akt (Ser473) (D9E) XP ® Rabbit mAb compared to a nonspecific negative control antibody (red). Western Blotting Protocol For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween ® 20 at 4°C with gentle shaking, overnight. NOTE: Please refer to primary antibody datasheet or product webpage for recommended antibody dilution. Solutions and Reagents From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: Western Blotting Application Solutions Kit NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water. • 20X Phosphate Buffered Saline (PBS): () To prepare 1 L 1X PBS: add 50 ml 20X PBS to 950 ml dH 2O, mix.