Human/Mouse/Rat HTRA2/Omi Antibody AF1458: R&D Systems

Human/Mouse/Rat HTRA2/Omi Antibody

(21 citations)   
  • Species Reactivity
    Human, Mouse, Rat
  • Specificity
    Detects human, mouse, and rat full length and mitochondria-processed HTRA2/Omi.
  • Source
    Polyclonal Rabbit IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human HTRA2/Omi
    Ala134-Glu458
    Accession # O43464
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.25 µg/mL
    See below
  • Simple Western
    2.5 µg/mL
    See below
  • Immunocytochemistry
    5-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples

Detection of Human/Mouse/Rat HTRA2/Omi by Western Blot. Western blot shows lysates of PC‑12 rat adrenal pheochromocytoma cell line, Jurkat human acute T cell leukemia cell line, HeLa human cervical epithelial carcinoma cell line, L‑929 mouse fibroblast cell line, and C2C12 mouse myoblast cell line. PVDF membrane was probed with 0.25 µg/mL of Rabbit Anti-Human/Mouse/Rat HTRA2/Omi Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1458) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Specific bands were detected for HTRA2/Omi at approximately 36 and 49 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.

Immunocytochemistry

HTRA2/Omi in Jurkat Human Cell Line. HTRA2/Omi was detected in immersion fixed Jurkat human acute T cell leukemia cell line stimulated with staurosporin using Rabbit Anti-Human/Mouse/Rat HTRA2/Omi Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1458) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (yellow; Catalog # NL004) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Human and Mouse HTRA2/Omi by Simple WesternTM. Simple Western lane view shows lysates of C2C12 mouse myoblast cell line and HeLa human cervical epithelial carcinoma cell line, loaded at 0.2 mg/mL. Specific bands were detected for HTRA2/Omi at approximately 51 kDa (precursor) and 41 kDa (processed) (as indicated) using 2.5 µg/mL of Rabbit Anti-Human/Mouse/Rat HTRA2/Omi Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1458). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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.
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. The specificity of the protease is yet to be defined and no endogenous substrates are known to date.

  • 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
  • Entrez Gene IDs:
    27429 (Human); 64704 (Mouse)
  • Alternate Names:
    High temperature requirement protein A2; HtrA serine peptidase 2; HTRA2; HtrA-like serine protease; Omi; OMIOmi stress-regulated endoprotease; PARK13EC 3.4.21.108; PRSS25; PRSS25serine, 25; Serine protease 25; serine protease HTRA2, mitochondrial; Serine proteinase OMI
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

21 Citations: Showing 1 - 10
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Species
Applications
Sample Type
  1. PARL mediates Smac proteolytic maturation in mitochondria to promote apoptosis
    Authors: S Saita, H Nolte, KU Fiedler, H Kashkar, AS Venne, RP Zahedi, M Krüger, T Langer
    Nat. Cell Biol, 2017;0(0):.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  2. Protease Omi cleaving Hax-1 protein contributes to OGD/R-induced mitochondrial damage in neuroblastoma N2a cells and cerebral injury in MCAO mice.
    Authors: Wu J, Li M, Cao L, Sun M, Chen D, Ren H, Xia Q, Tao Z, Qin Z, Hu Q, Wang G
    Acta Pharmacol Sin, 2015;36(9):1043-52.
    Species: Mouse
    Sample Type: Tissue Homogenates
    Application: WB
  3. Protease Omi facilitates neurite outgrowth in mouse neuroblastoma N2a cells by cleaving transcription factor E2F1.
    Authors: Ma Q, Hu Q, Xu R, Zhen X, Wang G
    Acta Pharmacol Sin, 2015;36(8):966-75.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  4. Heat shock inhibition of CDK5 increases NOXA levels through miR-23a repression.
    Authors: Morey T, Roufayel R, Johnston D, Fletcher A, Mosser D
    J Biol Chem, 2015;290(18):11443-54.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  5. PINK1 kinase catalytic activity is regulated by phosphorylation on serines 228 and 402.
    Authors: Aerts L, Craessaerts K, de Strooper B, Morais V
    J Biol Chem, 2015;290(5):2798-811.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  6. Influenza A virus protein PB1-F2 translocates into mitochondria via Tom40 channels and impairs innate immunity.
    Authors: Yoshizumi T, Ichinohe T, Sasaki O, Otera H, Kawabata S, Mihara K, Koshiba T
    Nat Commun, 2014;5(0):4713.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  7. The ubiquitin-conjugating enzymes UBE2N, UBE2L3 and UBE2D2/3 are essential for Parkin-dependent mitophagy.
    Authors: Geisler S, Vollmer S, Golombek S, Kahle P
    J Cell Sci, 2014;127(0):3280-93.
  8. Neural-specific deletion of Htra2 causes cerebellar neurodegeneration and defective processing of mitochondrial OPA1.
    Authors: Patterson, Victoria, Zullo, Alfred J, Koenig, Claire, Stoessel, Sean, Jo, Hakryul, Liu, Xinran, Han, Jinah, Choi, Murim, DeWan, Andrew T, Thomas, Jean-Leo, Kuan, Chia-Yi, Hoh, Josephin
    PLoS ONE, 2014;9(12):e115789.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
  9. p53-mediated activation of the mitochondrial protease HtrA2/Omi prevents cell invasion.
    Authors: Yamauchi S, Hou Y, Guo A, Hirata H, Nakajima W, Yip A, Yu C, Harada I, Chiam K, Sawada Y, Tanaka N, Kawauchi K
    J Cell Biol, 2014;204(7):1191-207.
    Species: Mouse
    Sample Type: Whole Cells
    Application: IF
  10. PINK1 is degraded through the N-end rule pathway.
    Authors: Yamano, Koji, Youle, Richard
    Autophagy, 2013;9(11):1758-69.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  11. The accumulation of misfolded proteins in the mitochondrial matrix is sensed by PINK1 to induce PARK2/Parkin-mediated mitophagy of polarized mitochondria.
    Authors: Jin, Seok Min, Youle, Richard
    Autophagy, 2013;9(11):1750-7.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  12. PINK1/Parkin-mediated mitophagy is dependent on VDAC1 and p62/SQSTM1.
    Authors: Geisler S, Holmstrom KM, Skujat D, Fiesel FC, Rothfuss OC, Kahle PJ, Springer W
    Nat. Cell Biol., 2010;12(2):119-31.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  13. Enhanced HtrA2/Omi expression in oxidative injury to retinal pigment epithelial cells and murine models of neurodegeneration.
    Authors: Ding X, Patel M, Shen D, Herzlich AA, Cao X, Villasmil R, Klupsch K, Tuo J, Downward J, Chan CC
    Invest. Ophthalmol. Vis. Sci., 2009;50(10):4957-66.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  14. Novel mitochondrial substrates of omi indicate a new regulatory role in neurodegenerative disorders.
    Authors: Johnson F, Kaplitt MG,
    PLoS ONE, 2009;4(9):e7100.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  15. Identification of a novel protein MICS1 that is involved in maintenance of mitochondrial morphology and apoptotic release of cytochrome c.
    Authors: Oka T, Sayano T, Tamai S, Yokota S, Kato H, Fujii G, Mihara K
    Mol. Biol. Cell, 2008;19(6):2597-608.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  16. HtrA2 regulates beta-amyloid precursor protein (APP) metabolism through endoplasmic reticulum-associated degradation.
    Authors: Huttunen HJ, Guenette SY, Peach C, Greco C, Xia W, Kim DY, Barren C, Tanzi RE, Kovacs DM
    J. Biol. Chem., 2007;282(38):28285-95.
    Species: Hamster
    Sample Type: Cell Lysates
    Application: WB
  17. Yersinia YopP-induced apoptotic cell death in murine dendritic cells is partially independent from action of caspases and exhibits necrosis-like features.
    Authors: Grobner S, Autenrieth SE, Soldanova I, Gunst DS, Schaller M, Bohn E, Muller S, Leverkus M, Wesselborg S, Autenrieth IB, Borgmann S
    Apoptosis, 2006;11(11):1959-68.
    Species: Mouse
    Sample Type: Cell Culture Supernates
    Application: WB
  18. Role and regulation of nodal/activin receptor-like kinase 7 signaling pathway in the control of ovarian follicular atresia.
    Authors: Wang H, Jiang JY, Zhu C, Peng C, Tsang BK
    Mol. Endocrinol., 2006;20(10):2469-82.
    Species: Rat
    Sample Type: Cell Lysates
    Application: WB
  19. Induction of BIM(EL) following growth factor withdrawal is a key event in caspase-dependent apoptosis of 661W photoreceptor cells.
    Authors: Gomez-Vicente V, Doonan F, Donovan M, Cotter TG
    Eur. J. Neurosci., 2006;24(4):981-90.
    Species: Mouse
    Sample Type: Whole Cells
    Application: ICC
  20. Roscovitine-induced up-regulation of p53AIP1 protein precedes the onset of apoptosis in human MCF-7 breast cancer cells.
    Authors: Wesierska-Gadek J, Gueorguieva M, Horky M
    Mol. Cancer Ther., 2005;4(1):113-24.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  21. Motoneuron resistance to apoptotic cell death in vivo correlates with the ratio between X-linked inhibitor of apoptosis proteins (XIAPs) and its inhibitor, XIAP-associated factor 1.
    Authors: Perrelet D, Perrin FE, Liston P, Korneluk RG, MacKenzie A, Ferrer-Alcon M, Kato AC
    J. Neurosci., 2004;24(15):3777-85.
    Species: Rat
    Sample Type: Tissue Homogenates
    Application: WB
Expand to show all 21 Citations
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