Sigma-Aldrich Anti-Ncf-1 Clone 2B8 Zoomab Rb Mono

Neutrophil cytosol factor 1 (UniProt: P14598; also known as NCF-1, 47 kDa autosomal chronic granulomatous disease protein, 47 kDa neutrophil oxidase factor, NCF-47K, Neutrophil NADPH oxidase factor 1, Nox organizer 2, Nox-organizing protein 2, SH3 and PX domain-containing protein 1A, p47-phox ) is encoded by the NCF1 (also known as NOXO2, SH3PXD1A ) gene (Gene ID: 653361) in human. NCF-1 is a subunit of the phagocyte NADPH oxidase complex that mediates the transfer of electrons from cytosolic NADPH to O2 to produce the superoxide anion (O2-). It is primarily localized in the cytosol, where it interacts with other components of the NADPH oxidase complex and, activation of NADPH-oxidase dependent superoxide generation requires the participation of several cytosolic factors including p47phox, p67phox, p40phox and Rac (1 or 2). NCF-1(p47-phox) is expressed in various tissues, with particularly high levels in immune cells such as neutrophils and macrophages. The protein has a N-terminal PX domain (aa 4-125aa) and two SH3 domains (aa 156-215 & aa 226-285). The PX domain mediates interaction with phosphatidylinositol 3,4-bisphosphate and other anionic phospholipids and the C-terminal SH3 domain precludes phospholipid binding and interaction with CYBA. Dysregulation of NCF-1 can lead to various diseases, particularly those involving immune dysfunction. Mutations in NCF1 (p47phox) cause autosomal recessive chronic granulomatous disease (CGD) with abnormal di-hydrorhodamine (DHR) assay and absent p47phox protein. CGD leads to impaired ROS production and increased susceptibility to infections. Recent studies have reported that NCF1-mediated oxidative-proinflammatory cascades and consequential pathological changes in the myometrium can play a crucial role in premature birth. This ZooMAb^® recombinant monoclonal antibody, generated by our proprietary technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Kuhns, D. B., et Al. (2019). Blood Adv. 3(2):136-147); Cheng, J., et Al. (2024). Sci Bulletin. 69(16):2604-2621).

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ZooMAb^® antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb^® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb^® antibodies are reliably available and ready to ship when you need them.

KLH-conjugated linear peptide corresponding to 22 amino acids from the internal region of human NCF-1.

Quality Control Testing

Evaluated by Western Blotting in RAW 264.7 cell lysate.

Western Blotting Analysis: A 1:10,000 dilution of this antibody detected NCF-1 in RAW 264.7 cell lysate.

Tested Applications

Western Blotting Analysis: A 1:10,000 dilution from a representative lot detected NCF-1 in lysates from Raji cells, Ramos cells, and rat spleen tissue.

Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected NCF-1 in Ramos cells.

Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected NCF-1 in human spleen tissue sections.

Affinity Binding Assay: A representative lot of this antibody bound NCF-1 peptide with at least one thousand-fold (1,000X) higher affinity than with non-specific control peptide.

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

Clone 2B8 is a ZooMAb^® recombinant rabbit monoclonal antibody that specifically detects NCF-1. It targets an epitope within 22 amino acids from the internal region.

Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS with 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 µL.

300 µg/mL after reconstitution at 25 µL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 µL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.

ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany

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.