Recombinant
RabMAb

Anti-HtrA2 / Omi antibody [EPR22] (ab75982)

Overview

  • Product name
    Anti-HtrA2 / Omi antibody [EPR22]
    See all HtrA2 / Omi primary antibodies
  • Description
    Rabbit monoclonal [EPR22] to HtrA2 / Omi
  • Tested applications
    Suitable for: WB, IP, IHC-P, ICC, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    A synthetic peptide corresponding to residues on human HtrA2

  • Positive control
    • Jurkat cell lysate, 293T cell lysate, HeLa cell lysate and Human colon tissue.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

Associated products

Applications

Our Abpromise guarantee covers the use of ab75982 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/500 - 1/5000. Detects a band of approximately 37 kDa (predicted molecular weight: 49 kDa).
IP 1/10.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC 1/250 - 1/500.
Flow Cyt 1/70.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • Function
    Serine protease that shows proteolytic activity against a non-specific substrate beta-casein. Promotes or induces cell death either by direct binding to and inhibition of BIRC proteins (also called inhibitor of apoptosis proteins, IAPs), leading to an increase in caspase activity, or by a BIRC inhibition-independent, caspase-independent and serine protease activity-dependent mechanism. Cleaves THAP5 and promotes its degradation during apoptosis. Isoform 2 seems to be proteolytically inactive.
  • Tissue specificity
    Isoform 1 is ubiquitous. Isoform 2 is expressed predominantly in the kidney, colon and thyroid.
  • Involvement in disease
    Defects in HTRA2 are the cause of Parkinson disease type 13 (PARK13) [MIM:610297]. A complex neurodegenerative disorder characterized by bradykinesia, resting tremor, muscular rigidity and postural instability, as well as by a clinically significant response to treatment with levodopa. The pathology involves the loss of dopaminergic neurons in the substantia nigra and the presence of Lewy bodies (intraneuronal accumulations of aggregated proteins), in surviving neurons in various areas of the brain.
  • Sequence similarities
    Belongs to the peptidase S1B family.
    Contains 1 PDZ (DHR) domain.
  • Domain
    The mature N-terminus is involved in the interaction with XIAP.
    The PDZ domain mediates interaction with MXI2.
  • Post-translational
    modifications
    Autoproteolytically activated.
  • Cellular localization
    Mitochondrion intermembrane space. Mitochondrion membrane. Predominantly present in the intermembrane space. Released into the cytosol following apoptotic stimuli, such as UV treatment, and stimulation of mitochondria with caspase-8 truncated BID/tBID.
  • Information by UniProt
  • Database links
  • Alternative names
    • High temperature requirement protein A2 antibody
    • HTRA 2 antibody
    • HtrA like serine protease antibody
    • HtrA serine peptidase 2 antibody
    • HtrA, E. coli, homolog of, 2 antibody
    • HtrA2 antibody
    • HTRA2_HUMAN antibody
    • mitochondrial antibody
    • Omi stress regulated endoprotease antibody
    • Omi stress-regulated endoprotease antibody
    • PARK 13 antibody
    • PARK13 antibody
    • Protease serine 25 antibody
    • PRSS 25 antibody
    • PRSS25 antibody
    • Serine protease 25 antibody
    • Serine protease HTRA2 antibody
    • Serine protease HTRA2 mitochondrial antibody
    • Serine protease htra2 mitochondrial precursor antibody
    • Serine protease omi antibody
    • Serine proteinase OMI antibody
    see all

Anti-HtrA2 / Omi antibody [EPR22] images

  • Immunohistochemical analysis of paraffin-embedded human colon using ab75982 at 1/100-1/250 dilution.
  • All lanes : Anti-HtrA2 / Omi antibody [EPR22] (ab75982) at 1/5000 dilution

    Lane 1 : Jurkat cell lysate
    Lane 2 : 293T cell lysate
    Lane 3 : HeLa cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size : 49 kDa
    Observed band size : 38 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 43 kDa. We are unsure as to the identity of these extra bands.

    Secondary antibody - goat anti-rabbit HRP (ab6721)

  • Overlay histogram showing HeLa cells stained with ab75982 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab75982, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was goat anti-rabbit Alexa Fluor® 488 (IgG H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

  • Anti-HtrA2 / Omi antibody [EPR22] (ab75982) at 1/1000 dilution + Isolated mitochondria from NB4 cell line at 30 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
    Developed using the ECL technique

    Predicted band size : 49 kDa
    Observed band size : 37 kDa (why is the actual band size different from the predicted?)


    Exposure time : 30 seconds

    This image is courtesy of an abreview submitted by Christian Marx

    Blocking buffer : 5% milk

    Diluting buffer : 1% milk

    High background signal after longer exposure.

    See Abreview

References for Anti-HtrA2 / Omi antibody [EPR22] (ab75982)

This product has been referenced in:
  • Miyamoto M  et al. High-temperature-required protein A2 as a predictive marker for response to chemotherapy and prognosis in patients with high-grade serous ovarian cancers. Br J Cancer 112:739-44 (2015). IHC-P ; Human . Read more (PubMed: 25628093) »
  • Mihailovich M  et al. miR-17-92 fine-tunes MYC expression and function to ensure optimal B cell lymphoma growth. Nat Commun 6:8725 (2015). WB ; Mouse . Read more (PubMed: 26555894) »

See all 3 Publications for this product

Product Wall

Application
Western blot
Sample
Mouse Cell lysate - whole cell (hepatocytes)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted Dec 11 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - other (isolated mitochondria from NB4 cell line)
Loading amount
30 µg
Specification
isolated mitochondria from NB4 cell line
Gel Running Conditions
Reduced Denaturing (13%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Christian Marx

Verified customer

Submitted Feb 15 2013

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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