|Application ||WB, IHC, FC, ICC, LCI|
|Reactivity||Human, Mouse, Rat|
|Calculated MW||41133 Da|
|Homology||Rat, mouse - identical.|
|Other Names||Proteinase-activated receptor 4, PAR-4, Coagulation factor II receptor-like 3, Thrombin receptor-like 3, F2RL3, PAR4|
|Related products for control experiments||Control 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.|
|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 Reconstitution||Lyophilized powder can be stored intact at room temperature for several weeks. For longer periods, it should be stored at -20°C.|
|Storage After Reconstitution||The 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 Reconstitution||Lyophilized 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.|
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Provided below are standard protocols that you may find useful for product applications.
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
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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