Human HTRA2/Omi Antibody (229926)

R&D Systems | Catalog # MAB1458

R&D Systems

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Western Blot, Immunocytochemistry

Cited:

Western Blot, Immunocytochemistry, ELISA Detection

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 229926
View all HTRA2/Omi Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human HTRA2/Omi
Ala134-Glu458
Accession # O43464

Specificity

Detects the mitochondria-processed form of human HTRA2/Omi.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Scientific Data Images for Human HTRA2/Omi Antibody (229926)

Detection of Human HTRA2/Omi antibody by Western Blot.

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

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

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

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

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

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

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.

Applications for Human HTRA2/Omi Antibody (229926)

Application
Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed K562 human chronic myelogenous leukemia cell line

Western Blot

1 µg/mL
Sample: 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

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: HTRA2/Omi

HTRA2/Omi is the mammalian homologue of bacterial high temperature requirement protein (HtrA). HTRA2/Omi localizes to the mitochondria and is processed to expose an amino-terminal Reaper-like motif similar to SMAC/Diablo. HTRA2/Omi is released from the mitochondria in response to apoptotic insult and can interact with the BIR2 or BIR3 domains of XIAP to relieve caspase-IAP inhibition. This effect can be measured by reversing XIAP-BIR2 (R&D Systems, Catalog # 786-XB) inhibition of Caspase-7 (R&D Systems, Catalog # 823-C7) cleavage of a fluorogenic peptide (DEVD-AFC, MP Bio, Catalog # AFC-138). IC50 values for this effect are typically between 0.2 and 1.5 μM. HTRA2/Omi is trimeric and functions as a serine protease. The serine protease activity may play a more central role in apoptosis than its IAP antagonizing function. A PDZ domain regulates the serine protease activity by blocking access to the active site.

References

  1. Suzuki, Y. et al. (2001) Mol. Cell. 8:613.
  2. van Loo, G. et al. (2002) Cell Death & Diff. 9:20.
  3. Hedge, R. et al. (2001) J. Biol. Chem. 277:432.
  4. Verhagen, A. et al. (2001) J. Biol. Chem. 277:445.
  5. Martins, L. et al. (2002) J. Biol. Chem. 277:439.
  6. Silke, J., and A. Verhagen (2002) Cell Death & Diff. 9:362.
  7. Savopoulos, J. et al. (2000) Protein Expression & Purification 19:227.

Long Name

High Temperature Requirement Protein-2

Alternate Names

Omi, PRSS25

Entrez Gene IDs

27429 (Human); 64704 (Mouse)

Gene Symbol

HTRA2

UniProt

O43464

Additional HTRA2/Omi Products

Product Documents for Human HTRA2/Omi Antibody (229926)

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Product Specific Notices for Human HTRA2/Omi Antibody (229926)

For research use only

Citations for Human HTRA2/Omi Antibody (229926)

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Protocols

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Associated Pathways

Apoptosis Signaling Pathway
Apoptosis Signaling Pathway Thumbnail