Skip to Content
MilliporeSigma
All Photos(1)

Key Documents

MABS1299

Sigma-Aldrich

Anti-PAR4 Antibody, clone 14H6

clone 14H6, from mouse

Synonym(s):

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

Sign Into View Organizational & Contract Pricing


About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41

biological source

mouse

Quality Level

antibody form

purified immunoglobulin

antibody product type

primary antibodies

clone

14H6, monoclonal

species reactivity

human

technique(s)

flow cytometry: suitable
immunocytochemistry: suitable
western blot: suitable

isotype

IgG2bκ

NCBI accession no.

UniProt accession no.

target post-translational modification

unmodified

Gene Information

human ... F2RL3(9002)

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).

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.

Immunogen

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

Application

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

Quality

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.

Target description

~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.

Physical form

Format: Purified

Other Notes

Concentration: Please refer to lot specific datasheet.

Not finding the right product?  

Try our Product Selector Tool.

Storage Class

12 - Non Combustible Liquids

wgk_germany

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

Already Own This Product?

Find documentation for the products that you have recently purchased in the Document Library.

Visit the Document Library

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

Contact Technical Service