nNOS Antibody

nNOS Polyclonal Antibody for Western blot, IF, ICC, IHC (P)

>> See 10 other antibodies for NOS1 (Neuronal Nitric Oxide Synthase, NOS Type 1, Brain NOS, bNOS1)
Synonyms:
Neuronal Nitric Oxide Synthase, NOS Type 1, Brain NOS, bNOS1
Entrez Gene ID:
Details
Host / Isotype: Rabbit
Class: Polyclonal
Type: Antibody
Tested Species Reactivity: Human (Hu), Mouse (Ms), Rat (Rt)
Published Species Reactivity: Rat (Rt)
Immunogen: Synthetic peptide corresponding to residues C N(1411) R L R S E S I A F I E E S K(1425) of human nNOS.
Ordering Information
Pierce nNOS Antibody
Product #
PA1-033
Size
100 µg
Price
$360.00
Purchase
Add nNOS Antibody to your cart.
Tested Applications Dilution *
Western Blot (WB) 1:100-1:1000
Immunofluorescence (IF) 1:50-1:500
Immunocytochemistry (ICC) 1:50-1:500
Immunohistochemistry (Paraffin) (IHC (P)) 1:20-1:200
Published Applications Dilution
Western Blot (WB) See publications below
Immunoprecipitation (IP) See publications below
* Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own experiment using appropriate negative and positive controls.
Form Information
Form: Liquid
Concentration: 1mg/ml
Purification: Antigen affinity chromatography
Storage Buffer: PBS with 1mg/ml BSA
Preservative: 0.05% sodium azide
Storage Conditions: -20° C, Avoid Freeze/Thaw Cycles
Product Specific Information
PA1-033 detects nNOS from human, rat and mouse samples.

PA1-033 has been successfully used in Western blot, ICC/IF and immunohistochemisty (paraffin) procedures. By Western blot, this antibody detects an ~160 kDa protein. Detects a few non-specific bands in Western blot analysis of rat and mouse brain lysates. This antibody is not recommended for MCF-7, PC-12, SK-N-C or U87-MG cells in Western blot applications.

The PA1-033 immunogen is a synthetic peptide to residues C N(1411) R L R S E S I A F I E E S K(1425) of human nNOS. The peptide sequence is 100% conserved in rat, mouse, frog, and rabbit nNos proteins. This peptide (Cat. # PEP-190) is available for use in neutralization and control experiments.
General Information
Nitric oxide (NO) is an inorganic, gaseous free radical that carries a variety of messages between cells. Vasorelaxation, neurotransmission and cytotoxicity can all be potentiated through cellular response to NO. NO production is mediated by members of the nitric oxide synthase (NOS) family. NOS catalyzes the oxidization of L-arginine to produce L-citrulline and NO. Two constitutive isoforms, brain or neuronal NOS (b or nNOS, type I) & endothelial cell NOS (eNOS, type III), and one inducible isoform (iNOS, type II), have been cloned. All NOS isoforms contain calmodulin, nicotinamide adenine dinucleotide phosphate (NADPH), flavin adenine dinucleotide (FAD), and flavin mononucleotide (FMN) binding domains.

iNOS is found in a variety of cell types including macrophages, hepatocytes, synoviocytes, and smooth muscle cells. Cytokines such as interferon-gamma (IFN), tumor necrosis factor (TNF), interleukin-1 and -2, and lipopolysaccarides (LPS) cause an increase in iNOS mRNA, protein, and activity levels. Protein kinase C-stimulating agents exhibit the same effect on iNOS activity. After cytokine induction, iNOS exhibits a delayed activity response which is then followed by a significant increase in NO production over a long period of time.
Product Images
  • nNOS Antibody (PA1-033) in WB
    PA1-033_WB_20130731155028.jpg

    Western Blot with anti-nNOS Polyclonal Antibody (PA1-033) (PA1-033_WB_20130731155028.jpg)

    Western Blot with anti-nNOS Polyclonal Antibody (PA1-033)

    Western blot analysis of nNOS was performed by loading 25 ug of mouse brain cell lysates onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4ºC overnight. The membrane was probed with a nNOS polyclonal antibody (Product # PA1-033) at a dilution of 1:500 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). Results show a band at approx. 160 kDa.
  • nNOS Antibody (PA1-033) in WB
    PA1-033_WB_rat_brain_20130808075614.jpg

    Western Blot with anti-nNOS Polyclonal Antibody (PA1-033) (PA1-033_WB_rat_brain_20130808075614.jpg)

    Western Blot with anti-nNOS Polyclonal Antibody (PA1-033)

    Western blot analysis of nNOS was performed by loading 25 ug of rat brain cell lysates onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4ºC overnight. The membrane was probed with a nNOS polyclonal antibody (Product # PA1-033) at a dilution of 1:200 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). Results show a band at approx. 160 kDa.
  • nNOS Antibody (PA1-033) in IF
    PA1-033_IF_U251_20131209140421.jpg

    Immunofluorescence with anti-nNOS Polyclonal Antibody (PA1-033) (PA1-033_IF_U251_20131209140421.jpg)

    Immunofluorescence with anti-nNOS Polyclonal Antibody (PA1-033)

    Immunofluorescent analysis of nNOS (green) showing staining in the cytoplasm and membrane of U251 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with an nNOS polyclonal antibody (Product # PA1-033) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.
  • nNOS Antibody (PA1-033) in IHC (P)
    PA1-033_IHC_Human_skeletal_muscle_tissue_20130613141632.jpg

    Immunohistochemistry (Paraffin) with anti-nNOS Polyclonal Antibody (PA1-033) (PA1-033_IHC_Human_skeletal_muscle_tissue_20130613141632.jpg)

    Immunohistochemistry (Paraffin) with anti-nNOS Polyclonal Antibody (PA1-033)

    Immunohistochemistry analysis of nNOS showing positive staining in the cytoplasm of paraffin-treated Human skeletal muscle (right) compared with a negative control in the absence of primary antibody (left). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a nNOS polyclonal antibody (Product # PA1-033) diluted by 3% BSA-PBS at a dilution of 1:200 overnight at 4°C in a humidified chamber. Tissues were washed extensively PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
  • nNOS Antibody (PA1-033) in IHC (P)
    PA1-033_IHC_Human_testis_tissue_20130613141701.jpg

    Immunohistochemistry (Paraffin) with anti-nNOS Polyclonal Antibody (PA1-033) (PA1-033_IHC_Human_testis_tissue_20130613141701.jpg)

    Immunohistochemistry (Paraffin) with anti-nNOS Polyclonal Antibody (PA1-033)

    Immunohistochemistry analysis of nNOS showing positive staining in the cytoplasm of paraffin-treated Human testis tissue (right) compared with a negative control in the absence of primary antibody (left). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a nNOS polyclonal antibody (Product # PA1-033) diluted by 3% BSA-PBS at a dilution of 1:200 overnight at 4°C in a humidified chamber. Tissues were washed extensively PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
  • nNOS Antibody (PA1-033) in IHC (P)
    PA1-033_IHC_Mouse_heart_tissue_20130613141739.jpg

    Immunohistochemistry (Paraffin) with anti-nNOS Polyclonal Antibody (PA1-033) (PA1-033_IHC_Mouse_heart_tissue_20130613141739.jpg)

    Immunohistochemistry (Paraffin) with anti-nNOS Polyclonal Antibody (PA1-033)

    Immunohistochemistry analysis of nNOS showing positive staining in the cytoplasm of paraffin-treated Mouse heart tissue (right) compared with a negative control in the absence of primary antibody (left). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a nNOS polyclonal antibody (Product # PA1-033) diluted by 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
Publications:
Western Blot
  Species / Dilution Summary
  Rt / 1:250
PA1-033 was used in western blot and immunoprecipitation to determine the nNOS splice variants in rat kidney and their roles in chronic kidney disease.

Nephrol Dial Transplant. 2009 May;24(5):1422-8.
"Splice variants of neuronal nitric oxide synthase are present in the rat kidney."
Author(s): Smith C, Merchant M, Fekete A, Nyugen HL, Oh P, Tain YL, Klein JB, Baylis C
Number of Citations: 1
(See PubMed article )
  Rt / 1:250
PA1-033 was used in western blot to investigate the renal nitric oxide production in rat pregnancy and possible involvement of different nitric oxide synthases

Am J Physiol Renal Physiol. 2010 Oct;299(4):F830-6.
"Renal nitric oxide production in rat pregnancy: role of constitutive nitric oxide synthases."
Author(s): Smith CA, Santymire B, Erdely A, Venkat V, Losonczy G, Baylis C
Number of Citations: 1
(See PubMed article )
  Rt / Not Cited
PA1-033 was used in western blot to investigate the therapeutic benefit of melatonin in hypertensive rats

J Pineal Res. 2010 Nov;49(4):390-8.
"Melatonin prevents hypertension and increased asymmetric dimethylarginine in young spontaneous hypertensive rats."
Author(s): Tain YL, Huang LT, Lin IC, Lau YT, Lin CY
Number of Citations: 2
(See PubMed article )
  Rt / Not Cited
PA1-033 was used in western blot to investigate the therapeutic effect of aliskiren for high blood pressure treatment

Eur J Pharmacol. 2011 Nov 30;670(2-3):561-5.
"Aliskiren prevents hypertension and reduces asymmetric dimethylarginine in young spontaneously hypertensive rats."
Author(s): Tain YL, Hsu CN, Lin CY, Huang LT, Lau YT
Number of Citations: 0
(See PubMed article )
Immunoprecipitation
  Species / Dilution Summary
  Rt / Not Cited
PA1-033 was used in western blot and immunoprecipitation to determine the nNOS splice variants in rat kidney and their roles in chronic kidney disease.

Nephrol Dial Transplant. 2009 May;24(5):1422-8.
"Splice variants of neuronal nitric oxide synthase are present in the rat kidney."
Author(s): Smith C, Merchant M, Fekete A, Nyugen HL, Oh P, Tain YL, Klein JB, Baylis C
Number of Citations: 1
(See PubMed article )
(This product is for In Vitro experimental use only. Not for resale without express authorization.)
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