Human HTRA2/Omi Antibody

Detection of Human HTRA2/Omi by Western Blot. Western blot shows lysates of SK-Mel-28 human malignant melanoma cell line, MCF-7 human breast cancer cell line, HEK293 human embryonic kidney cell line, and HeLa human cervical epithelial carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human HTRA2/Omi Monoclonal Antibody (Catalog # MAB1458) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for HTRA2/Omi at approximately 35 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4.

HTRA2/Omi in K562 Human Cell Line. HTRA2/Omi was detected in immersion fixed K562 human chronic myelogenous leukemia cell line using Mouse Anti-Human HTRA2/Omi Monoclonal Antibody (Catalog # MAB1458) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Detection of HTRA2/Omi by Western Blot HtrA2/Omi expression increases following CMV infection.(A–B) Immunoblot analyses of HtrA2/Omi in lysates of mock-infected HF (M) or HF infected (MOI of 3) with Towne-BAC or delta UL37x1 for 24, 48, 72, 96, or 120 h and control cell lysates from HeLa cells transfected with HtrA2/Omi expression plasmid (+) or control plasmid (−). The expected migration of the 49 kDa immature, pro-HtrA2/Omi and 36 kDA mature, HtrA2/Omi was calculated based on migration of molecular weight markers (not shown). Immunoblot detection of HtrA2, beta -actin and IE1 are shown. (C–N) Immunofluorescent images of HtrA2/Omi (C, F) and cytochrome c (I, L) (red), and GFP fluorescence (green) (D, G, J, M) and merged images (E, H, K, N) at 72 h (C–H) or 96 h (I–N) postinfection (MOI of 0.001) with delta UL37x1 (C–E, I–K) or Towne-BAC (F–H, L–N). Original magnification ×1000. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/18769594), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of HTRA2/Omi by Immunocytochemistry/ Immunofluorescence HtrA2/Omi expression increases following CMV infection.(A–B) Immunoblot analyses of HtrA2/Omi in lysates of mock-infected HF (M) or HF infected (MOI of 3) with Towne-BAC or delta UL37x1 for 24, 48, 72, 96, or 120 h and control cell lysates from HeLa cells transfected with HtrA2/Omi expression plasmid (+) or control plasmid (−). The expected migration of the 49 kDa immature, pro-HtrA2/Omi and 36 kDA mature, HtrA2/Omi was calculated based on migration of molecular weight markers (not shown). Immunoblot detection of HtrA2, beta -actin and IE1 are shown. (C–N) Immunofluorescent images of HtrA2/Omi (C, F) and cytochrome c (I, L) (red), and GFP fluorescence (green) (D, G, J, M) and merged images (E, H, K, N) at 72 h (C–H) or 96 h (I–N) postinfection (MOI of 0.001) with delta UL37x1 (C–E, I–K) or Towne-BAC (F–H, L–N). Original magnification ×1000. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/18769594), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of HTRA2/Omi by Immunocytochemistry/ Immunofluorescence Transient overexpression of HtrA2/Omi induces early death in CMV-infected but not uninfected cells.(A–F) Immunofluorescent images of HtrA2/Omi (A–C) and GFP fluorescence (D–F) following cotransfection of HF with GFP expression plasmid together with empty vector, HtrA2/Omi, or HtrA2S306A expression plasmids. Original magnification ×400. (G) Immunoblot depicting 49 kDa, immature and 36 kDA, mature forms of HtrA2/Omi and HtrA2S306A as well as beta -actin control in transfected HeLa cell lysates. (H) Percentage of live HFs at 120 h post cotransfection of GFP expression plasmid with vector, HtrA2/Omi, or HtrA2S306A expression plasmids. (I) Number of viral plaques 10 days following cotransfection of delta UL37x1 or Towne-BAC DNA (500 ng) with 800 ng vector, HtrA2/Omi, or HtrA2S306A expression plasmids. (J) Number of live (intact) cells in cultures at 48 or 72 h following transfection with Towne-BAC DNA (300 ng) alone or with 333 ng of HtrA2 or HtrA2S306A expression plasmids. (K) Number of live (intact) cells (±range) at 48 or 72 h post transfection with Towne-BAC DNA (500 ng) and 300 ng of vector or HtrA2/Omi expression plasmid with or without addition of 10 µM UCF-101 from 6 h. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/18769594), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of HTRA2/Omi by Western Blot HtrA2/Omi expression increases following CMV infection.(A–B) Immunoblot analyses of HtrA2/Omi in lysates of mock-infected HF (M) or HF infected (MOI of 3) with Towne-BAC or delta UL37x1 for 24, 48, 72, 96, or 120 h and control cell lysates from HeLa cells transfected with HtrA2/Omi expression plasmid (+) or control plasmid (−). The expected migration of the 49 kDa immature, pro-HtrA2/Omi and 36 kDA mature, HtrA2/Omi was calculated based on migration of molecular weight markers (not shown). Immunoblot detection of HtrA2, beta -actin and IE1 are shown. (C–N) Immunofluorescent images of HtrA2/Omi (C, F) and cytochrome c (I, L) (red), and GFP fluorescence (green) (D, G, J, M) and merged images (E, H, K, N) at 72 h (C–H) or 96 h (I–N) postinfection (MOI of 0.001) with delta UL37x1 (C–E, I–K) or Towne-BAC (F–H, L–N). Original magnification ×1000. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/18769594), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of HTRA2/Omi by Western Blot Transient overexpression of HtrA2/Omi induces early death in CMV-infected but not uninfected cells.(A–F) Immunofluorescent images of HtrA2/Omi (A–C) and GFP fluorescence (D–F) following cotransfection of HF with GFP expression plasmid together with empty vector, HtrA2/Omi, or HtrA2S306A expression plasmids. Original magnification ×400. (G) Immunoblot depicting 49 kDa, immature and 36 kDA, mature forms of HtrA2/Omi and HtrA2S306A as well as beta -actin control in transfected HeLa cell lysates. (H) Percentage of live HFs at 120 h post cotransfection of GFP expression plasmid with vector, HtrA2/Omi, or HtrA2S306A expression plasmids. (I) Number of viral plaques 10 days following cotransfection of delta UL37x1 or Towne-BAC DNA (500 ng) with 800 ng vector, HtrA2/Omi, or HtrA2S306A expression plasmids. (J) Number of live (intact) cells in cultures at 48 or 72 h following transfection with Towne-BAC DNA (300 ng) alone or with 333 ng of HtrA2 or HtrA2S306A expression plasmids. (K) Number of live (intact) cells (±range) at 48 or 72 h post transfection with Towne-BAC DNA (500 ng) and 300 ng of vector or HtrA2/Omi expression plasmid with or without addition of 10 µM UCF-101 from 6 h. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/18769594), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of HTRA2/Omi by Immunocytochemistry/ Immunofluorescence Transient overexpression of HtrA2/Omi induces early death in CMV-infected but not uninfected cells.(A–F) Immunofluorescent images of HtrA2/Omi (A–C) and GFP fluorescence (D–F) following cotransfection of HF with GFP expression plasmid together with empty vector, HtrA2/Omi, or HtrA2S306A expression plasmids. Original magnification ×400. (G) Immunoblot depicting 49 kDa, immature and 36 kDA, mature forms of HtrA2/Omi and HtrA2S306A as well as beta -actin control in transfected HeLa cell lysates. (H) Percentage of live HFs at 120 h post cotransfection of GFP expression plasmid with vector, HtrA2/Omi, or HtrA2S306A expression plasmids. (I) Number of viral plaques 10 days following cotransfection of delta UL37x1 or Towne-BAC DNA (500 ng) with 800 ng vector, HtrA2/Omi, or HtrA2S306A expression plasmids. (J) Number of live (intact) cells in cultures at 48 or 72 h following transfection with Towne-BAC DNA (300 ng) alone or with 333 ng of HtrA2 or HtrA2S306A expression plasmids. (K) Number of live (intact) cells (±range) at 48 or 72 h post transfection with Towne-BAC DNA (500 ng) and 300 ng of vector or HtrA2/Omi expression plasmid with or without addition of 10 µM UCF-101 from 6 h. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/18769594), licensed under a CC-BY license. Not internally tested by R&D Systems.