iNOS Recombinant Rabbit Monoclonal Antibody [PSH24-75] - BSA and Azide free
Catalog# HA751953
iNOS Recombinant Rabbit Monoclonal Antibody [PSH24-75] - BSA and Azide free
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WB
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IF-Cell
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FC(Intra)
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
iNOS Recombinant Rabbit Monoclonal Antibody [PSH24-75] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, FC(Intra)
Target Molecular Weight
Predicted band size: 130 kDa
Positive Control
Caco-2 (Human colorectal adenocarcinoma cell) cell lysate, RAW264.7 treated with 100ng/mL LPS for 24h cell lysate, NR8383 (Rat alveolar macrophage) cell lysate, NR8383 treated with 200ng/mL LPS for 4h add 1μg/mL BFA for last 2h cell lysate.
Conjugation
unconjugated
Clone Number
PSH24-75
Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage Buffer
PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:5,000
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IF-Cell
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1:1,000
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FC(Intra)
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1:10,000
Target
Function
Nitric oxide (NO) has a broad range of biological activities and has been implicated in signaling pathways in phylogenetically diverse species. Nitric oxide synthases (NOSs), the enzymes responsible for synthesis of NO, contain an N-terminal oxygenase domain and a C-terminal reductase domain. NOS activity requires homodimerization as well as three cosubstrates (L-arginine, NADPH and O2) and five cofactors or prosthetic groups (FAD, FMN, calmodulin, tetrahydrobiopterin and heme). Several distinct NOS isoforms have been described and been shown to represent the products of three distinct genes. These include two constitutive Ca2+/CaM-dependent forms of NOS, including NOS1 (also designated ncNOS) whose activity was first identified in neurons, and NOS3 (also designated ecNOS), first identified in endothelial cells. The inducible form of NOS, NOS2 (also designated iNOS), is Ca2+-independent and is expressed in a broad range of cell types.
Background References
1. Hokari A et al. Cloning and functional expression of human inducible nitric oxide synthase (NOS) cDNA from a glioblastoma cell line A-172. J Biochem 116:575-581 (1994).
2. Guo F H et al. Continuous nitric oxide synthesis by inducible nitric oxide synthase in normal human airway epithelium in vivo. Proc Natl Acad Sci U.S.A. 92:7809-7813 (1995).
Subcellular Location
Cytoskeleton, Nucleus, Cytosol.
Synonyms
HEP-NOS antibody
Hepatocyte NOS antibody
HEPNOS antibody
inducible antibody
Inducible nitric oxide synthase antibody
Inducible NO synthase antibody
Inducible NOS antibody
iNOS antibody
MAC NOS antibody
Macrophage NOS antibody
ExpandImages
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☑ Cell treatment (CT)
Western blot analysis of iNOS on different lysates with Rabbit anti-iNOS antibody (HA751953) at 1/5,000 dilution.
Lane 1: Caco-2 (Human colorectal adenocarcinoma cell) cell lysate
Lane 2: RAW264.7 (Mouse monocytic macrophage leukemia cell) cell lysate
Lane 3: RAW264.7 treated with 100ng/mL LPS for 24h cell lysate
Lysates/proteins at 20 µg/Lane.
Exposure time: 25 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA751953, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 130 kDa
Observed band size: 130 kDa -
☑ Cell treatment (CT)
Western blot analysis of iNOS on different lysates with Rabbit anti-iNOS antibody (HA751953) at 1/5,000 dilution.
Lane 1: NR8383 (Rat alveolar macrophage) cell lysate
Lane 2: NR8383 treated with 200ng/mL LPS for 4h add 1μg/mL BFA for last 2h cell lysate
Lysates/proteins at 20 µg/Lane.
Exposure time: 10 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA751953, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 130 kDa
Observed band size: 130 kDa -
☑ Cell treatment (CT)
Application: Immunocytochemistry (IF-cell)
Species: Mouse
Sample: RAW264.7 (Mouse monocytic macrophage leukemia cell) treated with 100ng/mL LPS for 24h
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.
Primary antibody: HA751953, 1/1,000, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue). -
☑ Cell treatment (CT)
Application: Flow Cytometry (Intra)
Species: Mouse
Sample: RAW264.7 (Mouse monocytic macrophage leukemia cell) treated with 100ng/mL LPS for 24h
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Tween-20, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.
Primary antibody: HA751953(1/10,000, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"