120-HD) for 15 minutes and MCF-7 human breast cancer cell line untreated or treated with 100 ng/mL Recombinant Human IGF-1 (291-G1) for 15 minutes. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-Akt (S473) Pan Specific Antigen Affinity-purified Polyclonal Antibody (Catalog # AF887), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). A specific band was detected for Phospho-Akt (S473) Pan Specific at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_lightbox"> 120-HD) for 15 minutes and MCF-7 human breast cancer cell line untreated or treated with 100 ng/mL Recombinant Human IGF-1 (291-G1) for 15 minutes. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-Akt (S473) Pan Specific Antigen Affinity-purified Polyclonal Antibody (Catalog # AF887), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). A specific band was detected for Phospho-Akt (S473) Pan Specific at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." alt="Western blot shows lysates of NIH-3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 100 ng/mL Human PDGF (120-HD) for 15 minutes and MCF-7 human breast cancer cell line untreated or treated with 100 ng/mL Recombinant Human IGF-1 (291-G1) for 15 minutes. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-Akt (S473) Pan Specific Antigen Affinity-purified Polyclonal Antibody (Catalog # AF887), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). A specific band was detected for Phospho-Akt (S473) Pan Specific at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_thumb" />

CTS005) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." class="big_lightbox"> CTS005) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." class="big_thumb" />

F0110). To facilitate intracellular staining, cells were fixed with formaldehyde (FC004) and permeabilized with Methanol." class="big_lightbox"> F0110). To facilitate intracellular staining, cells were fixed with formaldehyde (FC004) and permeabilized with Methanol." class="big_thumb" />

291-G1) for 15 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-Akt (S473) at approximately 60 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-Akt (S473) Pan Specific Antigen Affinity-purified Polyclonal Antibody (Catalog # AF887). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system." class="big_lightbox"> 291-G1) for 15 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-Akt (S473) at approximately 60 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-Akt (S473) Pan Specific Antigen Affinity-purified Polyclonal Antibody (Catalog # AF887). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system." class="big_thumb" />

HAF007). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4." class="big_lightbox"> HAF007). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4." alt="Western blot shows lysates of MCF-7 human breast cancer cell line, MBA-MB-123 human breast cancer cell line, and TS1 mouse helper T cell line. PVDF membrane was probed with 0.2 µg/mL Mouse Anti-Human/Mouse/Rat Akt Pan Specific Monoclonal Antibody (Catalog # MAB2055) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4." class="big_thumb" />

1775-KS), recombinant human Akt2, and recombinant human Akt3 (5 ng/lane). PVDF membrane was probed with 0.2 µg/mL Mouse Anti-Human/Mouse/Rat Akt Pan Specific Monoclonal Antibody (Catalog # MAB2055) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4." class="big_lightbox"> 1775-KS), recombinant human Akt2, and recombinant human Akt3 (5 ng/lane). PVDF membrane was probed with 0.2 µg/mL Mouse Anti-Human/Mouse/Rat Akt Pan Specific Monoclonal Antibody (Catalog # MAB2055) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4." class="big_thumb" />

NL007) and counter-stained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips." class="big_lightbox"> NL007) and counter-stained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips." class="big_thumb" />

MAB6815). Both antibodies were used at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to plasma membranes and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips." class="big_lightbox"> MAB6815). Both antibodies were used at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to plasma membranes and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips." class="big_thumb" />

VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in epithelial cells. Staining was performed using our IHC Staining with VisUCyte HRP Polymer Detection Reagents." class="big_lightbox"> VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in epithelial cells. Staining was performed using our IHC Staining with VisUCyte HRP Polymer Detection Reagents." class="big_thumb" />

VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in lymphocytes. Staining was performed using our IHC Staining with VisUCyte HRP Polymer Detection Reagents." class="big_lightbox"> VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in lymphocytes. Staining was performed using our IHC Staining with VisUCyte HRP Polymer Detection Reagents." class="big_thumb" />

MAB0041, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')₂Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin." class="big_lightbox"> MAB0041, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')₂Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin." class="big_thumb" />

222-AB) for 20 minutes. PVDF membrane was probed with 0.2 µg/mL of Mouse Anti-Human Phospho-Akt (S473) Pan Specific Monoclonal Antibody (Catalog # MAB887) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Phospho-Akt (S473) at approximately 65 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_lightbox"> 222-AB) for 20 minutes. PVDF membrane was probed with 0.2 µg/mL of Mouse Anti-Human Phospho-Akt (S473) Pan Specific Monoclonal Antibody (Catalog # MAB887) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Phospho-Akt (S473) at approximately 65 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." alt="Western blot shows lysates of Jurkat human acute T cell leukemia cell line, MCF-7 human breast cancer cell line, and NIH-3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 100 nm Calyculin for 30 minutes, 1 µg/mL insulin for 5 minutes, or 100 ng/mL Recombinant Human PDGF-AB (Catalog # 222-AB) for 20 minutes. PVDF membrane was probed with 0.2 µg/mL of Mouse Anti-Human Phospho-Akt (S473) Pan Specific Monoclonal Antibody (Catalog # MAB887) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Phospho-Akt (S473) at approximately 65 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_thumb" />

291-G1) for 20 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-Akt (S473) at approximately 65 kDa (as indicated) using 2 µg/mL of Mouse Anti-Human/Mouse Phospho-Akt (S473) Pan Specific Monoclonal Antibody (Catalog # MAB887). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system." class="big_lightbox"> 291-G1) for 20 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-Akt (S473) at approximately 65 kDa (as indicated) using 2 µg/mL of Mouse Anti-Human/Mouse Phospho-Akt (S473) Pan Specific Monoclonal Antibody (Catalog # MAB887). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system." class="big_thumb" />

HAF008). A specific band was detected for Akt Pan Specific, at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_lightbox"> HAF008). A specific band was detected for Akt Pan Specific, at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." alt="Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, Jurkat human acute T cell leukemia cell line, HepG2 human hepatocellular carcinoma cell line, MCF-7 human breast cancer cell line, and KG-1 human myeloid leukemia cell lines. PVDF membrane was probed with 0.1 µg/mL of Human Akt Pan Specific, Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2055) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Akt Pan Specific, at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_thumb" />