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Key Documents

MABS1255

Sigma-Aldrich

Anti-Platelet Factor 4 Antibody, clone 197.2

clone 197.2, from mouse

Sinónimos:

Platelet factor 4, C-X-C motif chemokine 4, Iroplact, Oncostatin-A, PF-4

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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

197.2, monoclonal

species reactivity

human

technique(s)

ELISA: suitable
flow cytometry: suitable
immunocytochemistry: suitable
western blot: suitable

isotype

IgG1κ

NCBI accession no.

UniProt accession no.

shipped in

ambient

target post-translational modification

unmodified

Gene Information

human ... PF4(5196)

General description

Platelet factor 4 (UniProt P02776; also known as C-X-C motif chemokine 4, Iroplact, Oncostatin-A, PF-4) is encoded by the PF4 (also known as CXCL4, SCYB4) gene (Gene ID 5196) in human. Platelet factor 4 (PF4) is a small CXC chemokine family cytokine released from alpha-granules of activated platelets during platelet aggregation to promote blood coagulation. PF4 binds heparin with high affinity (Kd ~4-20 nM) via its heparin-binding domain (a.a. 92-98) and neutralizes the anti-thrombin activity of heparin-like molecules. PF4 is chemotactic for neutrophils, fibroblasts and monocytes, and interacts with the chemokine receptor CXCR3 splice variant CXCR3B. In individuals suffering from heparin-induced thrombocytopenia (HIT), PF4-heparin complexes can trigger an immune response and the generation of anti-PF4 antibodies, resulting in the foramtion of macromolecular antibody-heparin-PF4 immune complexes that bind Fc RIIa (CD32a) on platelets and induce platelet activation. Platelet activation in turn results in more PF4 release into circulation, enhanced immune-complex formation, exaggerated platelet activation, and cell clearance. PF4 is synthesized with a signal peptide sequence (a.a. 1-31) that is removed posttranslationally to produce the mature PF4 (a.a. 32-101), additional proteolytic cleavage yields the PF4 short form (a.a. 48-101).

Specificity

Clone 197.2 targets a linear epitope sensitive to heparin binding, but insensitive to reducing agent or denaturation by SDS (Xiao, Z., et al. (2008). Blood. 112(4):1091-1100).

Immunogen

Purified human platelet PF4 (Xiao, Z., et al. (2008). Blood. 112(4):1091-1100).

Application

Detect PF4 using this mouse monoclonal Anti-Platelet Factor 4, clone 197.2, Cat. No. MABS1255, validated for use in ELISA, Flow Cytometry, Function Analysis, Immunocytochemistry. and Western Blotting.
Western Blotting Analysis: A representative lot detected E. coli expressed recombinant mature human PF4 (a.a. 32-101) under both reduced and non-reduced condition (Courtesy of Dr. Richard H. Aster, MD, PhD, Blood Center of Wisconsin, U. S. A.).

ELISA Analysis: A representative lot detected PF4 in the absence of heparin by ELISA. Decreased immunoreactivity of clone 197.2 was seen toward heparin-complexed PF4 (Xiao, Z., et al. (2008). Blood. 112(4):1091-1100).

Function Analysis: A representative lot and PF4, but not clone 197.2 or PF4 alone, activated human neutrophils as indicated by Mac-1 upregulation (Xiao, Z., et al. (2008). Blood. 112(4):1091-1100).

Immunocytochemistry Analysis: A representative lot, preconjugated with Alexa Fluor 488, immunolabeled the co-administered PF4 on human neutrophils. CD32a (Fc gamma RIIA) was seen co-localized with the PF4 anti-PF4 immune complexes on the surface of activated neutrophils (Xiao, Z., et al. (2008). Blood. 112(4):1091-1100).

Western Blotting Analysis: A representative lot detected PF4 under both reduced and non-reduced condition by recognizing a linear epitope (Xiao, Z., et al. (2008). Blood. 112(4):1091-1100).

Quality

Evaluated by Flow Cytometry in human platelets.

Flow Cytometry Analysis: 1 µg of this antibody detected PF4 in one million 4% paraformaldehyde-fixed, 0.2% Triton X-100-permeabilized human platelets.

Target description

~8 kDa (recombinant mature hPF4) observed. 10.84 kDa (pro-form), 7.769 kDa (mature; a.a. 32-101), 6.033 kDa (cleaved short form; a.a. 48-101) calculated. Uncharacterized bands may be observed in some lysate(s).

Physical form

Format: Purified
Purified mouse IgG1 in buffer containing PBS without preservatives.

Other Notes

Concentration: Please refer to lot specific datasheet.

Legal Information

ALEXA FLUOR is a trademark of Life Technologies

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Storage Class

12 - Non Combustible Liquids

wgk_germany

WGK 2

flash_point_f

Not applicable

flash_point_c

Not applicable


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