Revision 3

#72379

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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:
H M R

Sensitivity:
Endogenous

MW (kDa):
205

Source/Isotype:
Rabbit IgG

UniProt ID:
#Q86WI3

Entrez-Gene Id:
84166

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:50 - 1:250
Immunoprecipitation 1:100

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

NLRC5 (E1E9Y) Rabbit mAb recognizes endogenous levels of total NLRC5 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human NLRC5 protein.

Background

NLRC5 (NOD-like receptor family CARD domain containing 5, NOD4, CITA, NOD27) is a member of the NLR family of pattern recognition receptors (PRRs) involved in innate immunity (1,2). NLRC5 is predominantly expressed in hematopoietic cells, and is upregulated by IFN-γ (1-4). NLRC5 transcriptionally regulates the MHC class I antigen presentation pathway, and induces the expression of β2-microglobulin, TAP1, and LMP2 genes, in addition to MHC class I (3-5). In cancer, NLRC5 is critical for inducing cytotoxic CD8+ T cell activity and is a potential target for cancer immune evasion (6,7).

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 BSA, 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

H: Human M: Mouse R: Rat

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 3

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Western blot analysis of extracts from various cells and tissue using NLRC5 (E1E9Y) Rabbit mAb (upper) or α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). Negative expression of NLRC5 protein in K-562 and C2C12 cells is consistent with the predicted expression pattern. SDS-PAGE was performed using a 3-8% Tris-acetate gel.
Western Blotting Image 1: NLRC5 (E1E9Y) Rabbit mAb
Western blot analysis of extracts from HT-29 cells, untreated (-) or treated with Human Interferon-γ (hIFN-γ) Recombinant Protein #80385 (1,000 U/mL, 7 hr; +), using NLRC5 (E1E9Y) Rabbit mAb (upper) or α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). NLRC5 protein is induced by hIFN-γ treatment as expected. SDS-PAGE was performed using a 3-8% Tris-acetate gel.
Western Blotting Image 2: NLRC5 (E1E9Y) Rabbit mAb
Simple Western™ analysis of lysates (0.1 mg/mL) from SET-2 cells using NLRC5 (E1E9Y) Rabbit mAb #72379. The virtual lane view (left) shows the target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (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: NLRC5 (E1E9Y) Rabbit mAb
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 3

CST Logo
Immunoprecipitation of NLRC5 protein from SET-2 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is NLRC5 (E1E9Y) Rabbit mAb. Western blot analysis was performed using NLRC5 (E1E9Y) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody. SDS-PAGE was performed using a 3-8% Tris-acetate gel.
Immunoprecipitation Image 1: NLRC5 (E1E9Y) Rabbit mAb
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.