Revision 5

#62008

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Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, W-S, IP

Reactivity:
M

Sensitivity:
Endogenous

MW (kDa):
105

Source/Isotype:
Rabbit IgG

UniProt ID:
#Q9JJ26

Entrez-Gene Id:
54483

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunoprecipitation 1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity/Sensitivity

Pyrin/MEFV (F9C4G) Rabbit Monoclonal Antibody recognizes endogenous levels of total Pyrin/MEFV protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the carboxy terminus of mouse Pyrin/MEFV protein.

Background

Pyrin is encoded by the MEFV gene that is frequently mutated in patients with familial Mediterranean fever (FMF), an autoinflammatory disorder that causes recurrent fevers, usually also accompanied by pain in the abdomen, chest, and joints. It is expressed mainly in cells of the innate immune system, such as granulocytes, eosinophils, monocytes, and dendritic cells (1,2). Pyrin can form inflammasomes with ASC and Casp1, and this is regulated by RhoA guanosine triphosphatase (GTPase). RhoA activation leads to PKN1/2-mediated phosphorylation of Ser208 and Ser242 on Pyrin (Ser205 and Ser241 on mouse Pyrin, respectively). The chaperone 14-3-3 proteins bind to the phosphorylated Pyrin and keep it in an inactive state. Agents that inactivate RhoA, such as certain bacterial toxins, cause a decrease in PKN1/2 activity and Pyrin phosphorylation. This, in turn, releases Pyrin from the inhibitory 14-3-3 proteins and allows Pyrin to form an active inflammasome (3-6).

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

W: Western Blotting W-S: Simple Western™ IP: Immunoprecipitation

Cross-Reactivity Key

M: Mouse

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 5

Cell Signaling Technology Logo
Western blot analysis of extracts from various mouse cell lines and mouse peritoneal macrophages, untreated (-) or treated (+) as indicated with: Lipopolysaccharides #14011 (LPS; 1 μg/mL, 24 hr) and Mouse IL-4 Recombinant Protein #40004 (mIL-4; 12.5 ng/mL, 24 hr) using Pyrin/MEFV (F9C4G) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). Negative expression of Pyrin/MEFV protein in A20, C2C12, and Neuro-2a cells is consistent with the predicted expression pattern.
Western Blotting Image 1: Pyrin/MEFV (F9C4G) Rabbit Monoclonal Antibody
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length mouse Pyrin/MEFV protein (mPyrin/MEFV-Myc/DDK; +), using Pyrin/MEFV (F9C4G) Rabbit mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western Blotting Image 2: Pyrin/MEFV (F9C4G) Rabbit Monoclonal Antibody
Simple Western™ analysis of lysates (1 mg/mL) from mouse peritoneal macrophages treated with LPS (1 ug/mL; 24 hours ) and mIL-4 (12.5 ng/mL; 24 hours) using Pyrin/MEFV (F9C4G) Rabbit mAb #62008. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ ​​​​​​​ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Western Blotting Image 1: Pyrin/MEFV (F9C4G) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 5

Cell Signaling Technology Logo
Immunoprecipitation of Pyrin/MEFV protein from mouse peritoneal macrophage cell extracts. Cells were treated with Lipopolysaccharides #14011 (LPS; 1 μg/mL, 24 hr) and Mouse IL-4 Recombinant Protein #40004 (mIL-4; 12.5 ng/mL, 24 hr). Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotope Control #3900, and lane 3 is Pyrin/MEFV (F9C4G) Rabbit mAb. Western blot analysis was performed using Pyrin/MEFV (F9C4G) Rabbit mAb. Mouse Anti-Rabbit IgG (Light-Chain Specific) (D4W3E) mAb (HRP Conjugate) #93702 was used as a secondary antibody.
Immunoprecipitation Image 1: Pyrin/MEFV (F9C4G) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.