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Protease-activated Receptor-4 (extracellular) Antibody

Affinity purified rabbit polyclonal antibody

     
  • WB - Protease-activated Receptor-4 (extracellular) Antibody AG1057-050
    Western blot analysis of rat lung (lanes 1 and 3) and testis (lanes 2 and 4) lysates: 1, 2. Anti-Protease-Activated Receptor-4 (extracellular) antibody (#AG1057), (1:200). 3, 4. Anti-Protease-Activated Receptor-4 (extracellular) antibody, preincubated with the control peptide antigen.
  • WB - Protease-activated Receptor-4 (extracellular) Antibody AG1057-050
    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-Protease-Activated Receptor-4 (extracellular) antibody (#AG1057), (1:200). 4-6. Anti-Protease-Activated Receptor-4 (extracellular) antibody, preincubated with the control peptide antigen.
  • IHC - Protease-activated Receptor-4 (extracellular) Antibody AG1057-050
    Expression of PAR-4 in rat lung Immunohistochemical staining of rat lung paraffin-embedded sections using Anti-Protease-Activated Receptor-4 (extracellular) antibody (AG1057), (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.
  • IHC - Protease-activated Receptor-4 (extracellular) Antibody AG1057-050
    Expression of PAR-4 in rat epididymis Immunohistochemical staining of rat epididymis paraffin-embedded sections using Anti-Protease-Activated Receptor-4 (extracellular) antibody (AG1057), (1:100). Note that strong and specific staining is present in both the pseudostratified epithelium (blue arrow) and the smooth muscle cells of the muscular of blood vessels (red arrow). Hematoxilin is used as the counterstain.
  • ICC - Protease-activated Receptor-4 (extracellular) Antibody AG1057-050
    Expression of PAR-4 in a human breast adenocarcinoma cell line Immunocytochemical staining of a human breast adenocarcinoma cell line. A. Live intact human MCF-7 cells were stained with Anti-Protease-Activated Receptor-4 (extracellular) antibody (#AG1057), (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).
  • FC - Protease-activated Receptor-4 (extracellular) Antibody AG1057-050
    Anti-Protease-Activated_Receptor-4_(extracellular) - Indirect Flow cytometry analysis of human promyelocytic leukemia HL-60 cells: Indirect Flow cytometry analysis of human promyelocytic leukemia HL-60 cells:
    Black: Unstained cells + FITC-conjugated goat anti-rabbit antibody.
    Green: Cells + Anti-Protease-Activated Receptor-4 (extracellular) antibody (#APR-034), (1:40) + FITC conjugated goat anti-rabbit antibody.
  • SPECIFICATION
  • CITATIONS
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  • BACKGROUND
Product Information
Application
  • Applications Legend:
  • WB=Western Blot
  • IHC=Immunohistochemistry
  • IHC-P=Immunohistochemistry (Paraffin-embedded Sections)
  • IHC-F=Immunohistochemistry (Frozen Sections)
  • IF=Immunofluorescence
  • FC=Flow Cytopmetry
  • IC=Immunochemistry
  • ICC=Immunocytochemistry
  • E=ELISA
  • IP=Immunoprecipitation
  • DB=Dot Blot
  • CHIP=Chromatin Immunoprecipitation
  • FA=Fluorescence Assay
  • IEM=Immunoelectronmicroscopy
  • EIA=Enzyme Immunoassay
WB, IHC, FC, ICC, LCI
Primary Accession Q96RI0
Reactivity Human, Mouse, Rat
Host Rabbit
Clonality Polyclonal
Calculated MW 41133 Da
Homology Rat, mouse - identical.
Additional Information
Gene ID 9002
Other Names Proteinase-activated receptor 4, PAR-4, Coagulation factor II receptor-like 3, Thrombin receptor-like 3, F2RL3, PAR4
Related products for control experimentsControl peptide antigen (supplied with the antibody free of charge).
Target/Specificity 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.
Dilution WB~~1:200-1:2000
IHC~~1:100
ICC~~1:50
FC~~1:40
Peptide Confirmation Confirmed by amino acid analysis.
Application Details Western blot analysis (WB): - Rat DRGs (1:1000) (see Chen, D. et al. (2013) in Product Citations). Immunohistochemistry (IH): - Rat DRG sections (1:200) (see Chen, D. et al. (2013) in Product Citations). Immunocytochemistry (IC): - Rat dissociated DRGs (1:200) (see Chen, D. et al. (2013) in Product Citations).
Format Affinity purified antibody, lyophilized powder
Reconstitution 50 µl or 0.2 ml deionized water, depending on the sample size.
Antibody Concentration After Reconstitution 0.8 mg/ml.
Buffer After Reconstitution Phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.025% NaN3.
Storage Before ReconstitutionLyophilized powder can be stored intact at room temperature for several weeks. For longer periods, it should be stored at -20°C.
Storage After ReconstitutionThe reconstituted solution can be stored at 4ºC for up to 2 weeks. For longer periods, small aliquots should be stored at -20ºC or below. Avoid multiple freezing and thawing. The further dilutions should be made using a carrier protein such as BSA (1%). Centrifuge all antibody preparations before use (10000 × g 5 min).
Control Antigen Storage Before ReconstitutionLyophilized powder can be stored intact at room temperature for several weeks. For longer periods, it should be stored at -20°C.
Control Antigen Reconstitution 100 µl water.
Control Antigen Storage After Reconstitution-20ºC.
Preadsorption Control 1 µg peptide per 1 µg antibody.
Research Areas
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Background

Protease-activated receptor 4 (PAR-4) belongs to a family of four G protein-coupled receptors (PAR-1 - 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

References

1. MacFarlane, S.R. et al. (2001) Pharmacol. Rev. 53, 245.
2. Hollenberg, M.D. and Compton, S.J. (2002) Pharmacol. Rev. 54, 203.
3. Ossovskaya, V.S. and Bunnett, N.W. (2004) Physiol. Rev. 84, 579.

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Cat# AG1057-050
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