Anti-PAR4 (F2RL3) (extracellular) Antibody

Protease-activated receptor-4, PAR-4, Coagulation factor II receptor-like 3, Thrombin receptor-like 3
    Cat #: APR-034
    Alternative Name Protease-activated receptor-4, PAR-4, Coagulation factor II receptor-like 3, Thrombin receptor-like 3
  • KO Validated
  • Lyophilized Powder
  • Antigen Incl.
  • Type: Polyclonal
    Host: Rabbit
    Reactivity: h, m, r
    Immunogen
    • Peptide (C)HLRGQRWPFGEAA(S)R, corresponding to amino acid residues 136-150 of human PAR-4 (Accession Q96RI0). Cys 149 was replaced with Ser. 1st extracellular loop.
    • Anti-PAR4 (F2RL3) (extracellular) Antibody
    Accession (Uniprot) Number Q96RI0
    Gene ID 9002
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Rat, mouse - identical.
    RRID AB_2040089.
    Purity Affinity purified on immobilized antigen.
    Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
    Isotype Rabbit IgG.
    Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
    Reconstitution 50 μl or 0.2 ml double distilled water (DDW), depending on the sample size.
    Antibody concentration after reconstitution 0.8 mg/ml.
    Storage after reconstitution The reconstituted solution can be stored at 4°C for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 x g 5 min).
    Negative control antigen storage before reconstitution Lyophilized powder can be stored intact at room temperature for 2 weeks. For longer periods, it should be stored at -20°C.
    Negative control antigen reconstitution 100 µl double distilled water (DDW).
    Negative control antigen storage after reconstitution -20°C.
    Preadsorption Control 1 μg peptide per 1 μg antibody.
    Standard quality control of each lot Western blot analysis.
    Applications: ic, if, ifc, ih, lci, wb
    May also work in: ip*
    Western blot
    • Anti-PAR4 (F2RL3) (extracellular) Antibody
      Western blot analysis of rat lung (lanes 1 and 3) and testis (lanes 2 and 4) lysates:
      1,2. Anti-PAR4 (F2RL3) (extracellular) Antibody (#APR-034), (1:200).
      3,4. Anti-PAR4 (F2RL3) (extracellular) Antibody, preincubated with the negative control antigen.
    • Anti-PAR4 (F2RL3) (extracellular) Antibody
      Western blot analysis of human prostate carcinoma PC3 (lanes 1 and 4), and LNCaP (lanes 2 and 5), and human T cell leukemia Jurkat (lanes 3 and 6) cell lines:
      1-3. Anti-PAR4 (F2RL3) (extracellular) Antibody (#APR-034), (1:200).
      4-6. Anti-PAR4 (F2RL3) (extracellular) Antibody, preincubated with the negative control antigen.
    Immunohistochemistry
    • Anti-PAR4 (F2RL3) (extracellular) Antibody
      Expression of PAR-4 in rat lung
      Immunohistochemical staining of rat lung paraffin-embedded sections using Anti-PAR4 (F2RL3) (extracellular) Antibody (APR-034), (1:100). Note that staining is present in the respiratory epithelium of the bronchiole (blue arrow) as well as in the smooth muscle cells of the muscularis mucosae (red arrow). Hematoxilin is used as the counterstain.
    • Anti-PAR4 (F2RL3) (extracellular) Antibody
      Expression of PAR-4 in rat epididymis
      Immunohistochemical staining of rat epididymis paraffin-embedded sections using Anti-PAR4 (F2RL3) (extracellular) Antibody (APR-034), (1:100). Note that strong and specific staining is present in both the pseudostratified epithelium (blue arrow) and the smooth muscle cells of the blood vessels (red arrow). Hematoxilin is used as the counterstain.
    Indirect flow cytometry
    • Anti-PAR4 (F2RL3) (extracellular) Antibody
      Cell surface detection of PAR-4 in live human promyelocytic leukemia HL-60 cells:
      ___ Unstained cells + FITC-conjugated goat anti-rabbit antibody.
      ___ Cells + Anti-PAR4 (F2RL3) (extracellular) Antibody (#APR-034), (1:40) + FITC conjugated goat anti-rabbit antibody.
    • The negative control antigen is not suitable for this application.
    Live cell imaging / Immunocytochemistry
    • Anti-PAR4 (F2RL3) (extracellular) Antibody
      Expression of PAR-4 in a human breast adenocarcinoma cell line
      Cell surface detection of PAR-4 in a human breast adenocarcinoma cell line. A. Live intact human MCF-7 cells were stained with Anti-PAR4 (F2RL3) (extracellular) Antibody (#APR-034), (1:50), followed by goat anti-rabbit-AlexaFluor-488 secondary antibody (green). B. Live view of the cells. C. Nuclei were visualized with the cell-permeable dye Hoechst 33342 (blue).
    References
    1. MacFarlane, S.R. et al. (2001) Pharmacol. Rev. 53, 245.
    2. Hollenberg, M.D. et al. (2002) Pharmacol. Rev. 54, 203.
    3. Ossovskaya, V.S. et al. (2004) Physiol. Rev. 84, 579.
    Scientific background

    Protease-activated receptor 4 (PAR-4) belongs to a family of four G protein-coupled receptors (PAR1-4) that are activated as a result of proteolytic cleavage by certain serine proteases, hence their name. In this novel modality of activation, a specific protease cleaves the PAR receptor within a defined sequence in its extracellular N-terminal domain. This results in the creation of a new N-terminal tethered ligand, which subsequently binds to a site in the second extracellular loop of the same receptor. This binding results in the coupling of the receptor to G proteins and in the activation of several signal transduction pathways.1-3

    Different PARs are activated by different proteases. Hence, PAR-4 is activated by both thrombin and trypsin whereas PAR-1 and PAR-3 are activated only by thrombin and PAR-2 is activated only by trypsin.1-3 PAR-4 can be also cleaved and activated by other proteases such as cathepsin G.

    The intracellular signaling mechanisms mediated by PAR-4 activation are not completely elucidated but they involve calcium mobilization downstream of phospholipase Cβ through the Gαq pathway.1-3

    Tissue distribution of PAR-4 is very broad with the highest expression levels found in lung, testis, pancreas and small intestine. In addition, PAR-4 expression was observed in platelets, megakaryocytes and leukocytes. Studies with platelets derived form PAR-4 knockout mice have established an essential role for PAR-4 in thrombin-induced platelet activation. PAR-4 is likely involved in other physiological functions such as regulation of gastrointestinal motility and regulation of vascular endothelial cell function.1-3

    Application key:

    CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

    Species reactivity key:

    H- Human, M- Mouse, R- Rat
    Last update: 24/01/2020

    Anti-PAR4 (F2RL3) (extracellular) Antibody (#APR-034) is a highly specific antibody directed against an extracellular epitope of human protease-activated receptor-4. The antibody can be used in western blot analysis, immunohistochemical, immunocytochemical and flow cytometry applications. It has been designed to recognized PAR-4 from human, mouse and rat samples.

    For research purposes only, not for human use

    Applications

    Specifications

    Scientific Background

    Citations

    Citations
    KO validation citations
    1. Direct flow cytometry analysis of mouse platelets using #APR-034-F. Tested in PAR4-/- mice platelets.
      Arachiche, A. et al. (2013) PLoS ONE 8, e55740.
    Western blot citations
    1. Rat DRGs (1:1000).
      Chen, D. et al. (2013) Cell. Mol. Neurobiol. 33, 337.
    2. Rat DRGs.
      Wang, Z. et al. (2013) J. Neurosci. Res. 91, 1551.
    Immunohistochemistry citations
    1. Human colon sections.
      An, S. et al. (2017) Gut 66, 2040.
    2. Rat DRG sections (1:200).
      Chen, D. et al. (2013) Cell. Mol. Neurobiol. 33, 337.
    3. Rat DRG sections.
      Wang, Z. et al. (2013) J. Neurosci. Res. 91, 1551.
    Immunocytochemistry citations
    1. Rat dissociated DRGs (1:200).
      Chen, D. et al. (2013) Cell. Mol. Neurobiol. 33, 337.
    More product citations
    1. Sanchez-Centellas, D. et al. (2017) Thromb. Res. 154, 84.
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