WB result of iNOS Recombinant Rabbit mAb
Primary antibody: iNOS Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: Caco-2 whole cell lysate 40 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 131 kDa
Observed MW: 130 kDa
This blot was developed with high sensitivity substrate
iNOS Recombinant Rabbit mAb (S-2493-61)
iNOS Recombinant Rabbit mAb (S-2493-61)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | iNOS |
| Synonyms | Nitric oxide synthase, inducible; Hepatocyte NOS (HEP-NOS); Inducible NO synthase (Inducible NOS; iNOS); NOS type II; Peptidyl-cysteine S-nitrosylase NOS2; NOS2A; NOS2 |
| Immunogen | Synthetic Peptide |
| Location | Cytoplasm |
| Accession | P35228 |
| Clone Number | S-2493-61 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, ICC |
| Reactivity | Hu, Ms, Rt |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:1000 | Hu, Ms |
| IHC-P | 1:250 | Hu, Ms, Rt |
| ICC | 1:100 | Ms |
Background
Inducible nitric oxide synthase (iNOS) is a ~131 kDa homodimeric flavo-hemoprotein that, upon transcriptional up-regulation by cytokines such as IFN-γ, TNF-α and LPS via NF-κB and STAT pathways, catalyzes the NADPH- and O₂-dependent five-electron oxidation of L-arginine to generate large, sustained fluxes of nitric oxide (NO) and citrulline; this Ca²⁺-independent, calmodulin-bound enzyme localizes in cytosol and peroxisomes of macrophages, hepatocytes, chondrocytes and many other cell types, where the produced NO acts as a microbicidal, tumoricidal and immunomodulatory radical that can react with superoxide to form peroxynitrite, leading to protein tyrosine nitration, lipid peroxidation and DNA damage, thereby contributing to antimicrobial defense, vasodilatory shock in sepsis, inflammatory arthritis, neurotoxicity and carcinogenesis, while its expression is tightly controlled by transcriptional, post-transcriptional (mRNA stability, microRNAs) and post-translational (ubiquitin-proteasomal degradation, phosphorylation) mechanisms to balance beneficial and cytotoxic NO levels.
Picture
Picture
Western Blot
WB result of iNOS Recombinant Rabbit mAb
Primary antibody: iNOS Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: untreated RAW264.7 whole cell lysate 20 µg
Lane 2: RAW264.7 treated with 1 µg LPS for 6 hours whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 131 kDa
Observed MW: 130 kDa
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human cerebral cortex. Anti-iNOS antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human liver. Anti-iNOS antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human hepatocellular carcinoma. Anti-iNOS antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded mouse lung. Anti-iNOS antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded rat lung. Anti-iNOS antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Immunocytochemistry
ICC analysis of Raw264.7 cells treated with LPS (1 μg/mL, 6h) (top panel) and untreated Raw264.7 cells (below panel). Anti-iNOS antibody was used at 1/100 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
