Overview

  • Product nameAnti-PARL antibodySee all PARL primary antibodies ...
  • Description
    Rabbit polyclonal to PARL
  • Tested applicationsWB, ICC/IF, IHC-P more details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat, Cow, Orangutan
  • Immunogen

    Synthetic peptide corresponding to Human PARL aa 350 to the C-terminus conjugated to Keyhole Limpet Haemocyanin (KLH).
    (Peptide available as ab45230)

  • Positive control
    • ab42638 gave a positive result in Jurkat Whole cell Lysate and Hela Whole Cell Lysate (data not shown). This antibody gave a positive result in IHC in the following FFPE tissue: Human colon adenocarcinoma.

Properties

Applications

Our Abpromise guarantee covers the use of ab42638 in the following tested applications.

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

Application Notes
WB Use a concentration of 1 µg/ml. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).Can be blocked with Human PARL peptide (ab45230).
ICC/IF Use a concentration of 5 µg/ml.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • FunctionRequired for the control of apoptosis during postnatal growth. Essential for proteolytic processing of an antiapoptotic form of OPA1 which prevents the release of mitochondrial cytochrome c in response to intrinsic apoptoptic signals (By similarity). Promotes changes in mitochondria morphology regulated by phosphorylation of P-beta domain.
  • Sequence similaritiesBelongs to the peptidase S54 family.
  • Post-translational
    modifications
    P-beta is proteolytically processed (beta-cleavage) in a PARL-dependent manner. The cleavage is inhibited when residues Ser-65, Thr-69 and Ser-70 are all phosphorylated.
  • Cellular localizationMitochondrion inner membrane and Nucleus. Translocated into the nucleus by an unknown mechanism.
  • Target information above from: UniProt accession Q9H300 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links
  • Alternative names
    • Mitochondrial intramembrane cleaving protease PARL antibody
    • Mitochondrial intramembrane-cleaving protease PARL antibody
    • P-beta antibody
    • parl antibody
    • PARL_HUMAN antibody
    • Pbeta antibody
    • presenilin associated rhomboid-like antibody
    • Presenilins associated rhomboid-like protein antibody
    • PRO2207 antibody
    • PSARL antibody
    • PSARL1 antibody
    • PSENIP2 antibody
    • RHBDS1 antibody
    • Rhomboid 7 homolog 1 antibody
    see all

Anti-PARL antibody images

  • IHC image of PARL staining in Human colon adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab42638, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

     

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Lane 1 : Anti-PARL antibody (ab42638) at 1 µg/ml
    Lane 2 : Anti-PARL antibody (ab42638)

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 42 kDa
    Observed band size : 37 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 17 kDa (possible non-specific binding).

    Exposure time : 2 minutes

    This antibody was raised against an immunogen that is predicted to recognize isoforms 1 and 2 of Presenilins-associated rhomboid-like protein. The predicted molecular weights of isoforms 1 and 2 are 42kDa and 37kDa respectively.

  • ICC/IF image of ab42638 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab42638, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

  • All lanes : Anti-PARL antibody (ab42638) at 1/1000 dilution

    Lane 1 : Lysate prepared from mitochondria isolated from HEK293T cells transfected with non-targeting siRNA.
    Lane 2 : Lysate prepared from mitochondria isolated from HEK293T cells transfected with PARL-specific siRNA.

    Lysates/proteins at 24 µg per lane.

    Secondary
    Goat anti-rabbit IgG conjugated to HRP at 1/5000 dilution
    developed using the ECL technique

    Predicted band size : 42 kDa
    Observed band size : 30 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 32 kDa,33 kDa,38 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 10 seconds

    Image courtesy of Andrew Greene by Abreview.

    Cytochrome C was used as a loading control. The low molecular weight of the PARL band may be due to N-terminal cleavage (reported by Sik et al. JBC 2003).

    See Abreview

References for Anti-PARL antibody (ab42638)

ab42638 has not yet been referenced specifically in any publications.

Product Wall

Application Western blot
Sample Human Cell lysate - other (Mitochondria isolated from HEK293T cells)
Loading amount 24 µg
Specification Mitochondria isolated from HEK293T cells
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Mr. Andrew Greene

Verified customer

Submitted May 31 2011

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