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Anti-PAR4 Antibody, clone 5F10，100UL，MABS1298，Sigma

销售价： ¥ 4290.40 / 件

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订货号： 0BW0581

品牌型号： Sigma MABS1298

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最小订货量： 1件

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包装参数

产品介绍 Product Description

General description

Proteinase-activated receptor 4 (UniProt Q96RI0; also known as Coagulation factor II receptor-like 3, PAR-4, Thrombin receptor-like 3) is encoded by the F2RL3 (also known as PAR4) gene (Gene ID 9002) in human. Protease-activated receptors (PARs) constitute a unique family of seven-transmembrane, G-protein-coupled receptors (GPCRs) activated by proteolytic cleavage of their N-terminal propeptide sequence. Once cleaved off, the N-terminal propeptide fragment functions as a ligand and activates the receptor by binding the second extracellular loop. The four PAR family members (PAR-1 to PAR-4) are widely expressed and activated by multiple proteases, and utilize different types of G-proteins (Gi, Gq, and G12/13) for signal transdution depending on the activating protease and cellular context. PAR-4 is expressed on platelets and exhibits a low-affinity for thrombin. However, PAR-4 is able to form hetero-oligomers with both PAR-1 and the ADP receptor P2Y12 to mediate thrombin- and ADP-initiated signaling. PAR-4 cleavage is significantly enhanced through hetero-oligomerization with PAR-1, and PAR-4 interaction with P2Y12 is directly linked to arrestin-2 recruitment and AKT signaling. PAR-4 is a 7-transmembrane (a.a. 83-103, 109-129, 152-172, 192-213, 248-268, 284-304, 320-343) GPCR activated by thrombin cleavage between R47 and G48, having 3 extracellular loops and 3 intracellular loops between the extracellular N-terminal end (a.a. 48-82) the cytoplasmic C-terminal tail (a.a. 344-385).
~45 kDa observed. 41.13/39.16 kDa (prepro-/pro-PAR4) calculated. The broad banding pattern and larger apparent band size is consistent with the detection of glycosylated PAR4. Uncharacterized band(s) may appear in some lysates.

Immunogen

Epitope: Near (N-terminal to) the thrombin cleavage site.
MBP-conjugated recombinant human PAR4 N-terminal fragment including the propeptide sequence.

Application

Anti-PAR4 Antibody, clone 5F10 is an antibody against PAR4 for use in Western Blotting, Immunocytochemistry, Flow Cytometry, Inhibition.
Immunocytochemistry Analysis: 20 µg/mL from a representative lot detected endogenous PAR4 by fluorescent immunocytochemistry staining of freshly isolated human platelets (Courtesy of Dr. Marvin Nieman, Case Western Reserve University, Cleveland, OH, USA).
Immunocytochemistry Analysis: A representative lot detected the expression of exogenously transfected human PAR4 by fluorescent immunocytochemistry staining of 4% formaldehyde-fixed HEK293 Flp-In cells following tetracycline treatment (Mumaw, M.M., et al. (2015). Thromb. Res. 135(6):1165-1171).
Flow Cytometry Analysis: A representative lot detected tetracycline-induced expression of exogenously transfected human PAR4 on the surface of HEK293 Flp-In cells. Thrombin treatment diminished cell surface PAR4 immunoreactivity (Mumaw, M.M., et al. (2015). Thromb. Res. 135(6):1165-1171).
Inhibition Analysis: A representative lot, when added prior to thrombin, protected cell surface PAR4 against thrombin cleavage (Mumaw, M.M., et al. (2015). Thromb. Res. 135(6):1165-1171).
Western Blotting Analysis: A representative lot detected MBP fusion proteins containing human PAR4 fragment a.a. 18-78 or 41-66, but not 48-72. MBP-PAR4 fusion degradation by thrombin treatment ablolished target band detection by clone 5F10 (Mumaw, M.M., et al. (2015). Thromb. Res. 135(6):1165-1171).
Western Blotting Analysis: A representative lot detected tetracycline-induced expression of exogenously introduced human PAR4 in a HEK293 Flp-In cell line, as well as endogenous PAR4 in isolated human platelets (hPLTs). Thrombin activation of hPLTs diminished PAR4 target band detection (Mumaw, M.M., et al. (2015). Thromb. Res. 135(6):1165-1171).
Research Category
Signaling
Research Sub Category
GPCR, cAMP/cGMP & Calcium Signaling

Biochem/physiol Actions

Clone 5F10 recognizes a propeptide epitope near (N-terminal to) the thrombin-cleavage site and protects cell surface PAR4 against thrombin cleavage (Mumaw, M.M., et al. (2015). Thromb. Res. 135(6):1165-1171). Clone 5F10 recognizes prepro- and pro-, but not proteolytically activated, forms of human PAR4.

Physical form

Format: Purified
Protein G purified.
Purified mouse monoclonal IgG2bλ antibody in PBS without preservatives.

Preparation Note

Stable for 1 year at -20°C from date of receipt.

Analysis Note

Evaluated by Western Blotting in human platelet lysate.

Western Blotting Analysis: A 1:250 dilution of this antibody detected PAR4 in 50 µg of human platelet lysate.

Other Notes

Concentration: Please refer to lot specific datasheet.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

技术参数 Specifications

biological source	mouse
Quality Level	100
conjugate	unconjugated
antibody form	purified immunoglobulin
antibody product type	primary antibodies
clone	5F10, monoclonal
species reactivity	human
technique(s)	flow cytometry: suitable,immunocytochemistry: suitable,inhibition assay: suitable,western blot: suitable
isotype	IgG2bλ
NCBI accession no.	NP_003941
UniProt accession no.	Q96RI0
target post-translational modification	unmodified
Gene Information	human ... F2RL3(9002)

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