Anti-PAR4, clone 14H6 (mouse monoclonal)

Species Reactivity	Key Applications	Host	Format	Antibody Type
H	WB, ICC, FC	M	Purified	Monoclonal Antibody

Species Reactivity

Key Applications

Host

Format

Antibody Type

WB, ICC, FC

Purified

Monoclonal Antibody

Description
Catalogue Number	MABS1299
Description	Anti-PAR4 Antibody, clone 14H6
Alternate Names	Proteinase-activated receptor 4 PAR-4 Coagulation factor II receptor-like 3 Thrombin receptor-like 3
Background Information	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).

Description

Catalogue Number

MABS1299

Description

Anti-PAR4 Antibody, clone 14H6

Alternate Names

Proteinase-activated receptor 4
PAR-4
Coagulation factor II receptor-like 3
Thrombin receptor-like 3

Background Information

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).

Product Information
Format	Purified
Presentation	Purified mouse monoclonal IgG2bκ antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Product Information

Format

Purified

Presentation

Purified mouse monoclonal IgG2bκ antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Applications
Application	Anti-PAR4 Antibody, clone 14H6 is an antibody against PAR4 for use in Western Blotting, Immunocytochemistry, Flow Cytometry.
Key Applications	Western Blotting Immunocytochemistry Flow Cytometry
Application Notes	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). Immunocytochemistry Analysis: A representative lot detected endogenous PAR4 by fluorescent immunocytochemistry staining of 4% formaldehyde-fixed human platelets. Thrombin treatment diminished PAR4 immunoreactivity (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). Western Blotting Analysis: A representative lot detected MBP fusion proteins containing human PAR4 fragment a.a. 18-78, 41-66, or 48-72. MBP-PAR4 fusion cleavage by thrombin ablolished target band detection by clone 14H6 (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).

Applications

Application

Anti-PAR4 Antibody, clone 14H6 is an antibody against PAR4 for use in Western Blotting, Immunocytochemistry, Flow Cytometry.

Key Applications

Western Blotting
Immunocytochemistry
Flow Cytometry

Application Notes

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).
Immunocytochemistry Analysis: A representative lot detected endogenous PAR4 by fluorescent immunocytochemistry staining of 4% formaldehyde-fixed human platelets. Thrombin treatment diminished PAR4 immunoreactivity (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).
Western Blotting Analysis: A representative lot detected MBP fusion proteins containing human PAR4 fragment a.a. 18-78, 41-66, or 48-72. MBP-PAR4 fusion cleavage by thrombin ablolished target band detection by clone 14H6 (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).

Biological Information
Immunogen	MBP-conjugated recombinant human PAR4 N-terminal fragment including the propeptide sequence.
Epitope	Near (C-terminal to) the thrombin cleavage site.
Clone	14H6
Concentration	Please refer to lot specific datasheet.
Host	Mouse
Specificity	Clone 14H6 recognizes an epitope near (C-terminal) to the thrombin-cleavage site. Unlike clone 5F10 (Cat. No. MABS1298), clone 14H6 does not protect cell surface PAR4 against thrombin cleavage (Mumaw, M.M., et al. (2015). Thromb. Res. 135(6):1165-1171). Clone 14H6 recognizes prepro- and pro-, but not proteolytically activated, human PAR4. A structual change at the cleavage site following thrombin cleavage is believed to prevent the detection of the activated PAR-4 by clone 5F10.
Isotype	IgG2bκ
Species Reactivity	Human
Species Reactivity Note	Human.
Antibody Type	Monoclonal Antibody
Entrez Gene Number	NP_003941
Gene Symbol	F2RL3 PAR4
Purification Method	Protein G Purified
UniProt Number	Q96RI0
Molecular Weight	~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.

Biological Information

Immunogen

MBP-conjugated recombinant human PAR4 N-terminal fragment including the propeptide sequence.

Epitope

Near (C-terminal to) the thrombin cleavage site.

Clone

14H6

Concentration

Please refer to lot specific datasheet.

Host

Mouse

Specificity

Clone 14H6 recognizes an epitope near (C-terminal) to the thrombin-cleavage site. Unlike clone 5F10 (Cat. No. MABS1298), clone 14H6 does not protect cell surface PAR4 against thrombin cleavage (Mumaw, M.M., et al. (2015). Thromb. Res. 135(6):1165-1171). Clone 14H6 recognizes prepro- and pro-, but not proteolytically activated, human PAR4. A structual change at the cleavage site following thrombin cleavage is believed to prevent the detection of the activated PAR-4 by clone 5F10.

Isotype

IgG2bκ

Species Reactivity

Human

Species Reactivity Note

Human.

Antibody Type

Monoclonal Antibody

Entrez Gene Number

NP_003941

Gene Symbol

F2RL3
PAR4

Purification Method

Protein G Purified

UniProt Number

Q96RI0

Molecular Weight

~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.

Product Usage Statements
Quality Assurance	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.
Usage Statement	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.

Product Usage Statements

Quality Assurance

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.

Usage Statement

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.

Storage and Shipping Information
Storage Conditions	Stable for 1 year at 2-8°C from date of receipt.

Storage and Shipping Information

Storage Conditions

Stable for 1 year at 2-8°C from date of receipt.

Packaging Information
Material Size	100 µL

Packaging Information

Material Size

100 µL

实验耗材

分子生物学试剂

蛋白与免疫学

生化试剂

细胞及检测试剂

色谱及层析

仪器设备

实验服务

办公用品

Anti-PAR4, clone 14H6 (mouse monoclonal)

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