Product Datasheet. AKT1 [p Ser473] Antibody NB Unit Size: 0.1 mg. Store at -20C. Avoid freeze-thaw cycles. Publications: 3

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1 Product Datasheet AKT1 [p Ser473] Antibody NB Unit Size: 0.1 mg Store at -20C. Avoid freeze-thaw cycles. Publications: 3 Protocols, Publications, Related Products, Reviews, Research Tools and Images at: Updated 9/6/2018 v.20.1 Earn rewards for product reviews and publications. Submit a publication at Submit a review at

2 NB AKT1 [p Ser473] Antibody Product Information Unit Size Concentration Storage Clonality Preservative Isotype Purity 0.1 mg 1 mg/ml Store at -20C. Avoid freeze-thaw cycles. Polyclonal 0.01% Sodium Azide IgG Immunogen affinity purified Buffer 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, ph 7.2 Product Description Host Rabbit Gene ID 207 Gene Symbol Species Reactivity Notes Specificity/Sensitivity Immunogen Product Application Details Applications AKT1 Human, Mouse, Rat Reactivity against AKT1 from other species may occur, but has not yet been tested AKT [Ser473] This whole rabbit serum was prepared by repeated immunizations with a synthetic peptide corresponding to a C-terminus region near aa of human, mouse, rat and chicken AKT1 proteins conjugated to KLH. Western Blot, ELISA, Flow Cytometry, Immunocytochemistry/Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin Recommended Dilutions Western Blot 1:200-1:1000, Flow Cytometry 1:10-1:1000, ELISA 1: :60000, Immunohistochemistry 1:100-1:500, Immunocytochemistry/Immunofluorescence 1:10-1:500, Immunohistochemistry- Paraffin 1:10-1:500 Application Notes Images For immunohistochemistry no pre-treatment of sample is required. Western Blot: Akt1 [p Ser473] Antibody [NB ] - Lane 1: AKT1 Recombinant Protein. Lane 2: AKT1 Mutant Human Recombinant Protein. Lane 3: AKT1 (phosphatase treated) Human Recombinant Protein. Load: 50 ng per lane. Primary antibody: AKT ps473 antibody at 1:1,000 for overnight at 4C. Secondary antibody: Peroxidase rabbit secondary antibody at 1:40,000 for 30 min at RT. Block: Blocking Buffer for Fluorescent Western Blotting for 30 min at RT. Predicted/Observed size: 56 kda, 56 kda for Gli-2. Other band(s): Gli-2 splice variants and isoforms. Page 1 of 4 v.20.1 Updated 9/6/2018

3 Immunocytochemistry/Immunofluorescence: Akt1 [p Ser473] Antibody [NB ] - Tissue: cardiomyocytes infected with adenovirus expressing with wild-type AKT. Fixation: 0.5% PFA. Antigen retrieval: not required. Primary antibody: AKT ps473 antibody at 1:40 for 1 h at RT. Secondary antibody: texas-red conjugated rabbit secondary antibody at 1:10,000 for 45 min at RT. Localization: AKT ps473 is nuclear. Staining: AKT ps473 as green fluorescent signal with texas-red conjugated phalloidin (red) to label filamentous actin. Page 2 of 4 v.20.1 Updated 9/6/2018 Immunohistochemistry: Akt1 [p Ser473] Antibody [NB ] - Tissue: human breast carcinoma. Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required. Primary antibody: Akt ps473 antibody at 100 dilution for 1 h at RT. Secondary antibody: Dako's Techmate streptavidin-biotin reagents at 1:10,000 for 45 min at RT. Localization: Akt ps473 is nuclear and occasionally cytoplasmic. Western Blot: Akt1 [p Ser473] Antibody [NB ] - Lane 1: nuclear extract from cells infected with adenovirus expressing nuclear-targeted AKT kinase. Load: 35 ug per lane. Primary antibody: AKT ps473 antibody at 1:200 dilution for overnight at 4C. Secondary antibody: IRDye800 rabbit secondary antibody at 1:10,000 for 45 min at RT. Block: 5% BLOTTO overnight at 4C. Predicted/Observed size: 56 kda for AKT ps473. Other band(s): unspecific. Immunohistochemistry-Paraffin: Akt1 [p Ser473] Antibody [NB ] - Rabbit anti-akt ps473 was used at a 1:100 dilution to detect AKT in human breast tumor tissue. The staining is much stronger than the weak basal level of phosphorylation in normal breast.signal was developed using Dako's Techmate streptavidinbiotin reagents. Tissue was formalinfixed and paraffin embedded.

4 Immunohistochemistry-Paraffin: Akt1 [p Ser473] Antibody [NB ] - Rabbit used at a 1:100 dilution to detect AKT in normal human breast tissue. AKT is weakly phosphorylated in normal tissue in the breast. The phosphorylated AKT is clearly localized in the cytoplasm.signal was developed using Dako's Techmate streptavidin-biotin reagents. Tissue was formalin-fixed and paraffin embedded. Page 3 of 4 v.20.1 Updated 9/6/2018 Publications Jung CK, Jung SH, Yim SH et al. Predictive micrornas for lymph node metastasis in endoscopically resectable submucosal colorectal cancer. Oncotarget. May :00AM [PMID: ] (IHC, Human) Cuesto G, Jordan-alvarez S, Enriquez-Barreto L et al. GSK3B Inhibition Promotes Synaptogenesis in Drosophila and Mammalian Neurons. PLoS ONE Mar 13 [PMID: ] (WB, Human) Ansell SM, Tang H, Kurtin PJ et al. Temsirolimus and rituximab in patients with relapsed or refractory mantle cell lymphoma: a phase 2 study. Lancet Oncol Mar 24. [PMID: ]

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