PA3-031A

antibody from Invitrogen Antibodies

Targeting: NOS3 ECNOS, eNOS

Provider product page for PA3-031A

Western blot

Immunocytochemistry

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [28]
Comments [0]
Validations
Immunocytochemistry [2]
Immunohistochemistry [3]
Other assay [6]

Submit

Product number: PA3-031A - Provider product page
Provider: Invitrogen Antibodies
Product name: eNOS Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Description: PA3-031A detects endothelial Nitric Oxide Synthase (eNOS) from canine, bovine, human, mouse and rat tissues. This antibody does not detect brain NOS (bNOS) or inducible NOS (iNOS). PA3-031A has been successfully used in Western blot, immunohistochemistry, immunofluorescence, and immunocytochemistry procedures. By Western blot, this antibody detects an approximately 140 kDa band representing eNOS from canine cardiac extracts. Immunohistochemical staining of eNOS in mouse cardiac tissue with PA3-031 results in the staining of the endothelium. The PA3-031A immunogen is a synthetic peptide corresponding to residues P(599) Y N S S P R P E Q H K S Y K(613) C of bovine eNOS.
Reactivity: Human, Mouse, Rat, Bovine, Canine
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Concentration: Conc. Not Determined
Storage: -20° C, Avoid Freeze/Thaw Cycles

NOS3 protein structure - PA3-031A shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on cancer biopsies of deparaffinized Human lung adenocarcinoma tissues. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a rabbit polyclonal antibody recognizing eNOS (Product # PA3-031A) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on normal deparaffinized Human placenta tissue tissues. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:500 with a rabbit polyclonal antibody recognizing eNOS (Product # PA3-031A) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on normal deparaffinized Human tonsil tissue tissues. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a rabbit polyclonal antibody recognizing eNOS (Product # PA3-031A) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 3 Immunostaining (A) for eNOS in the adrenal cortex (zona fasciculata) and Optical density (B) of the adrenal cells (zona fasciculata). (A): Negative control (a), healthy normal-weight dogs (b), dogs with HAC (c). Brown cytoplasm and nuclei indicate positive staining, whereas blue cytoplasm and nuclei are negative (haematoxylin). Arrows indicate nuclear staining. The scale bars are 50 (top figures) and 20 mum (bottom figures). (B): Optical Healthy normal-weight dogs present cells with a higher optical density (greater immunostaining) than dogs with HAC (**P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 10.1371/journal.pone.0251129.g002 Fig 2 Laminar flow increases NO production in GEnCs. mGEnCs were cultured either under static conditions or under a laminar flow of 5 dyn/cm2 for 7 days. Representative images of stainings for NO (probed with DAF-FM diacetate) ( A ), iNOS ( E ) and eNOS ( G ) of mGEnC cultured under static (left panel) or flow (right panel). Arrows indicate the direction of the flow applied. Quantification of the fluorescent intensity of NO ( B ), iNOS ( F ) and eNOS ( H ) staining, respectively. mRNA expression levels of iNOS ( C ) and eNOS ( D ) in mGEnCs cultured under either static conditions or flow. * p

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 7 ( a ), ( d ) Immunohistochemistry staining images showing expression of eNOS in cardiac muscle x100 (inset: x400) and thoracic aorta x400 respectively of control and tramadol treated mice. Arrows show the specific staining for the antibody. ( b ), ( e ) Relative quantification of immunohistochemistry staining for eNOS in cardiac muscle and thoracic aorta, respectively of both groups, n = 10. Data are expressed as means +- SEM, *p < 0.05 for control versus tramadol. ( c ), ( f ) Relative quantitative expression of mRNA levels of eNOS in cardiac muscle and thoracic aorta respectively of control and tramadol treated mice. All gene expression levels were normalized to GAPDH, n = 10. Data are expressed as means +- SEM, *p < 0.05 for control versus tramadol. ( g ) Immunohistochemistry staining images x100 (inset: x400) showing VEGF expression in cardiac muscle of control and tramadol treated mice. Arrows show the specific staining for the antibody. ( h ) Relative quantification of immunohistochemistry staining for VEGF in cardiac muscle of control and tramadol treated mice, n = 10. Data are expressed as means +- SEM, *p < 0.05 for control versus tramadol.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2. nNOS expression. (A) Immunohistochemical localization of nNOS expression of the testicular tissue and semiquantitative results of nNOS content obtained by densitometric analysis of immunohistochemistry. (B) Immunohistochemical localization of iNOS expression of the testicular tissue and semiquantitative results of iNOS content obtained by densitometric analysis of immunohistochemistry. (C) Immunohistochemical localization of eNOS expression of the testicular tissue and semiquantitative results of eNOS content obtained by densitometric analysis of immunohistochemistry. (I) Group I (control), (II) Group II (STZ-induced diabetic rats), (III) Group III (STZ-induced diabetic rats +50 mg/kg/day PTX through the experiment), (IV) Group IV (STZ-induced diabetic +50 mg/kg/day PTX for just second month), (V) Group V (only 50 mg/kg/day PTX administered to rats), and (NC) Negative controls. nNOS (neuronal nitric oxide synthase), iNOS (inducible nitric oxide synthase), eNOS (endothelial nitric oxide synthase), STZ (streptozotosin) PTX (pentoxifylline). * p < 0.05 compared to Group I, ** p < 0.05 compared to Group II.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 4 Combined re-expression of ASS1 and ASL suppresses growth in vivo. A Combined doxycycline-induced ectopic expression of ASS1+ ASL in 786-O cells. HSP90 is used as a loading control. B 786-O subcutaneous xenograft tumor growth in nude mice, where cells have doxycycline induced expression of ASS1 and ASL or empty vector ( n = 6). C Immunohistochemistry staining and quantification for phosphorylated histone H3 as a marker for proliferation in xenograft tumors. Scale bars are at 100 mum. D Immunohistochemistry staining and quantification for cleaved caspase 3 as a marker for apoptosis in xenograft tumors. Scale bars are at 100 mum. ** p < 0.01. E Immunohistochemistry staining and quantification for p21 as a marker for proliferation in xenograft tumors. Scale bars are at 100 mum. *** p < 0.001. F Nitric oxide metabolite levels in matched ccRCC tumors and normal kidney as measured using gas-phase chemiluminescence ( n = 20). G Nitric oxide synthase activity measured by radiolabeled citrulline generation, plotted as a fold change between matched normal and ccRCC tumor tissues ( n = 20). H Representative immunohistochemistry images with quantification for NOS1,2,3 protein expression in matched tumor-normal pairs (Biomax OD-CT-UrKid03-002) ( n = 31). Scale bars represent 200 mum. **** p < 0.001, paired t -test

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 6 Digital images of the immunohistochemical expression of inducible nitric oxide synthase (photomicrography, x 200). A: SO; B: SO + G; C: PPVL; D: PPVL + G. SO: Control group; PPVL: Partial portal vein ligation; G: Glutamine; iNOS: Inducible nitric oxide synthase.