HTRA2/Omi Antibody [Unconjugated]

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.

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.

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.

• Reactivity: Hu, Mu, Rt
• Applications: WB, Simple Western, ICC/IF
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: LYOPH

HTRA2/Omi Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human HTRA2/Omi
  Ala134-Glu458
  Accession # O43464
• Specificity: Detects human, mouse, and rat full length and mitochondria-processed HTRA2/Omi.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: HTRA2
• Purity: Immunogen affinity purified
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 5-15 ug/mL
  • Simple Western 2.5 ug/mL
  • Western Blot 0.25 ug/mL
• Application Notes: In Simple Western only 10-15 uL of the recommended dilution is used per data point.

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Purity: Immunogen affinity purified
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for HTRA2/Omi Antibody [Unconjugated]

• 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

Background

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.

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for HTRA2/Omi Antibody (AF1458)(23)

Publications using AF1458

• R Aishwarya, S Alam, CS Abdullah, M Morshed, SS Nitu, M Panchatcha, S Miriyala, CG Kevil, MS Bhuiyan Pleiotropic effects of mdivi-1 in altering mitochondrial dynamics, respiration, and autophagy in cardiomyocytes Redox Biol, 2020-07-26;36(0):101660. 2020-07-26 [PMID: 32750667] (Western Blot, Rat)
• ÉM Szeg?, A Dominguez-, E Gerhardt, A König, DJ Koss, W Li, R Pinho, C Fahlbusch, M Johnson, P Santos, A Villar-Piq, T Thom, S Rizzoli, M Schmitz, J Li, I Zerr, J Attems, O Jahn, TF Outeiro Cytosolic Trapping of a Mitochondrial Heat Shock Protein Is an Early Pathological Event in Synucleinopathies Cell Rep, 2019-07-02;28(1):65-77.e6. 2019-07-02 [PMID: 31269451] (Western Blot, Rat)
• S Saita, H Nolte, KU Fiedler, H Kashkar, AS Venne, RP Zahedi, M Krüger, T Langer PARL mediates Smac proteolytic maturation in mitochondria to promote apoptosis Nat. Cell Biol, 2017-03-13;0(0):. 2017-03-13 [PMID: 28288130] (Western Blot, Human)
• Wu J, Li M, Cao L, Sun M, Chen D, Ren H, Xia Q, Tao Z, Qin Z, Hu Q, Wang G Protease Omi cleaving Hax-1 protein contributes to OGD/R-induced mitochondrial damage in neuroblastoma N2a cells and cerebral injury in MCAO mice. Acta Pharmacol Sin, 2015-08-24;36(9):1043-52. 2015-08-24 [PMID: 26299953] (Western Blot, Mouse)
• Ma Q, Hu Q, Xu R, Zhen X, Wang G Protease Omi facilitates neurite outgrowth in mouse neuroblastoma N2a cells by cleaving transcription factor E2F1. Acta Pharmacol Sin, 2015-08-01;36(8):966-75. 2015-08-01 [PMID: 26238290] (Western Blot, Human, Mouse)
• Morey T, Roufayel R, Johnston D, Fletcher A, Mosser D Heat shock inhibition of CDK5 increases NOXA levels through miR-23a repression. J Biol Chem, 2015-03-31;290(18):11443-54. 2015-03-31 [PMID: 25829494] (Western Blot, Human)
• 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 Neural-specific deletion of Htra2 causes cerebellar neurodegeneration and defective processing of mitochondrial OPA1. PLoS ONE, 2014-12-22;9(12):e115789. 2014-12-22 [PMID: 25531304] (Western Blot, Mouse)
• Aerts L, Craessaerts K, de Strooper B, Morais V PINK1 kinase catalytic activity is regulated by phosphorylation on serines 228 and 402. J Biol Chem, 2014-12-19;290(5):2798-811. 2014-12-19 [PMID: 25527497] (Western Blot, Human)
• Yoshizumi T, Ichinohe T, Sasaki O, Otera H, Kawabata S, Mihara K, Koshiba T Influenza A virus protein PB1-F2 translocates into mitochondria via Tom40 channels and impairs innate immunity. Nat Commun, 2014-08-20;5(0):4713. 2014-08-20 [PMID: 25140902] (Western Blot, Human)
• Geisler S, Vollmer S, Golombek S, Kahle P The ubiquitin-conjugating enzymes UBE2N, UBE2L3 and UBE2D2/3 are essential for Parkin-dependent mitophagy. J Cell Sci, 2014-06-06;127(0):3280-93. 2014-06-06 [PMID: 24906799] (ICC, Human)
• Yamauchi S, Hou Y, Guo A, Hirata H, Nakajima W, Yip A, Yu C, Harada I, Chiam K, Sawada Y, Tanaka N, Kawauchi K p53-mediated activation of the mitochondrial protease HtrA2/Omi prevents cell invasion. J Cell Biol, 2014-03-24;204(7):1191-207. 2014-03-24 [PMID: 24662565] (IHC, Mouse)
• Jin , Seok Min, Youle , Richard The accumulation of misfolded proteins in the mitochondrial matrix is sensed by PINK1 to induce PARK2/Parkin-mediated mitophagy of polarized mitochondria. Autophagy, 2013-09-05;9(11):1750-7. 2013-09-05 [PMID: 24149988] (Western Blot, Human)
• Yamano , Koji, Youle , Richard PINK1 is degraded through the N-end rule pathway. Autophagy, 2013-04-17;9(11):1758-69. 2013-04-17 [PMID: 24121706] (Western Blot, Human)
• Geisler S, Holmstrom KM, Skujat D, Fiesel FC, Rothfuss OC, Kahle PJ, Springer W PINK1/Parkin-mediated mitophagy is dependent on VDAC1 and p62/SQSTM1. Nat. Cell Biol., 2010-01-24;12(2):119-31. 2010-01-24 [PMID: 20098416] (ICC, Human)
• Johnson F, Kaplitt MG Novel mitochondrial substrates of omi indicate a new regulatory role in neurodegenerative disorders. PLoS ONE, 2009-09-18;4(9):e7100. 2009-09-18 [PMID: 19763263] (Western Blot, Human)
• Ding X, Patel M, Shen D, Herzlich AA, Cao X, Villasmil R, Klupsch K, Tuo J, Downward J, Chan CC Enhanced HtrA2/Omi expression in oxidative injury to retinal pigment epithelial cells and murine models of neurodegeneration. Invest. Ophthalmol. Vis. Sci., 2009-05-14;50(10):4957-66. 2009-05-14 [PMID: 19443712] (ICC, IHC-Fr, Human, Mouse)
• Oka T, Sayano T, Tamai S, Yokota S, Kato H, Fujii G, Mihara K Identification of a novel protein MICS1 that is involved in maintenance of mitochondrial morphology and apoptotic release of cytochrome c. Mol. Biol. Cell, 2008-04-16;19(6):2597-608. 2008-04-16 [PMID: 18417609] (ICC, Human)
• Huttunen HJ, Guenette SY, Peach C, Greco C, Xia W, Kim DY, Barren C, Tanzi RE, Kovacs DM HtrA2 regulates beta-amyloid precursor protein (APP) metabolism through endoplasmic reticulum-associated degradation. J. Biol. Chem., 2007-08-06;282(38):28285-95. 2007-08-06 [PMID: 17684015] (ICC, Western Blot, Hamster)
• Grobner S, Autenrieth SE, Soldanova I, Gunst DS, Schaller M, Bohn E, Muller S, Leverkus M, Wesselborg S, Autenrieth IB, Borgmann S Yersinia YopP-induced apoptotic cell death in murine dendritic cells is partially independent from action of caspases and exhibits necrosis-like features. Apoptosis, 2006-11-01;11(11):1959-68. 2006-11-01 [PMID: 17036200] (Western Blot, Mouse)
• Gomez-Vicente V, Doonan F, Donovan M, Cotter TG Induction of BIM(EL) following growth factor withdrawal is a key event in caspase-dependent apoptosis of 661W photoreceptor cells. Eur. J. Neurosci., 2006-08-01;24(4):981-90. 2006-08-01 [PMID: 16930425] (ICC, Mouse)
• Wang H, Jiang JY, Zhu C, Peng C, Tsang BK Role and regulation of nodal/activin receptor-like kinase 7 signaling pathway in the control of ovarian follicular atresia. Mol. Endocrinol., 2006-05-18;20(10):2469-82. 2006-05-18 [PMID: 16709598] (Western Blot, Rat)
• Wesierska-Gadek J, Gueorguieva M, Horky M Roscovitine-induced up-regulation of p53AIP1 protein precedes the onset of apoptosis in human MCF-7 breast cancer cells. Mol. Cancer Ther., 2005-01-01;4(1):113-24. 2005-01-01 [PMID: 15657359] (ICC, Human)
• Perrelet D, Perrin FE, Liston P, Korneluk RG, MacKenzie A, Ferrer-Alcon M, Kato AC 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. J. Neurosci., 2004-04-14;24(15):3777-85. 2004-04-14 [PMID: 15084658] (Western Blot, Rat)

Reviews for HTRA2/Omi Antibody (AF1458) (3)

Average Rating: 4.3

(Based on 3 reviews)

We have 3 reviews tested in 2 species: Human, Pig.

reviewed by: L P ICC Pig 06/07/2017

Summary

• Application: Immunocytochemistry
• Sample Tested: pig SC cells
• Species: Pig
• Lot: N/A

reviewed by: L P ICC Pig 06/07/2017

Summary

• Application: Immunocytochemistry
• Sample Tested: Hep2G cells
• Species: Pig
• Lot: N/A

reviewed by: Anonymous - IF Human 01/05/2015

Summary

• Application: Immunofluorescence
• Sample Tested: See PMID 23183826
• Species: Human

Bioinformatics

• Gene Symbol: HTRA2
• Uniprot:
  • O43464()