- PA1-033 - Invitrogen Antibodies NOS1 antibody

Submitted references TGFβR-1/ALK5 inhibitor RepSox induces enteric glia-to-neuron transition and influences gastrointestinal mobility in adult mice.

Shi CJ, Lian JJ, Zhang BW, Cha JX, Hua QH, Pi XP, Hou YJ, Xie X, Zhang R
Acta pharmacologica Sinica 2023 Jan;44(1):92-104

ASS1 and ASL suppress growth in clear cell renal cell carcinoma via altered nitrogen metabolism.

Khare S, Kim LC, Lobel G, Doulias PT, Ischiropoulos H, Nissim I, Keith B, Simon MC
Cancer & metabolism 2021 Dec 3;9(1):40

Reduced Basal Nitric Oxide Production Induces Precancerous Mammary Lesions via ERBB2 and TGFβ.

Ren G, Zheng X, Bommarito M, Metzger S, Walia Y, Letson J, Schroering A, Kalinoski A, Weaver D, Figy C, Yeung K, Furuta S
Scientific reports 2019 Apr 30;9(1):6688

Comparative synaptosome imaging: a semi-quantitative method to obtain copy numbers for synaptic and neuronal proteins.

Richter KN, Wildhagen H, Helm MS, Ußling JE, Schikorski T, Rizzoli SO
Scientific reports 2018 Oct 4;8(1):14838

Aging-dependent decrease in the numbers of enteric neurons, interstitial cells of Cajal and expression of connexin43 in various regions of gastrointestinal tract.

Sun T, Li D, Hu S, Huang L, Sun H, Yang S, Wu B, Ji F, Zhou D
Aging 2018 Dec 11;10(12):3851-3865

Effects of Aloe Vera on Spinal Cord Ischemia-Reperfusion Injury of Rats.

Yuksel Y, Guven M, Kaymaz B, Sehitoglu MH, Aras AB, Akman T, Tosun M, Cosar M
Journal of investigative surgery : the official journal of the Academy of Surgical Research 2016 Dec;29(6):389-398

Pregnant rats treated with a high-fat/prooxidant Western diet with ANG II and TNF-α are resistant to elevations in blood pressure and renal oxidative stress.

Cunningham MW Jr, West CA, Wen X, Deng A, Baylis C
American journal of physiology. Regulatory, integrative and comparative physiology 2015 Jun 1;308(11):R945-56

RNA silencing targeting PIN (protein inhibitor of neuronal nitric oxide synthase) attenuates the development of hypertension in young spontaneously hypertensive rats.

Wang SC, Lin KM, Chien SJ, Huang LT, Hsu CN, Tain YL
Journal of the American Society of Hypertension : JASH 2014 Jan;8(1):5-13

Renal nitric oxide synthase and antioxidant preservation in Cyp1a1-Ren-2 transgenic rats with inducible malignant hypertension.

Cunningham MW Jr, Sasser JM, West CA, Milani CJ, Baylis C, Mitchell KD
American journal of hypertension 2013 Oct;26(10):1242-9

Aminoguanidine attenuates hypertension, whereas 7-nitroindazole exacerbates kidney damage in spontaneously hypertensive rats: the role of nitric oxide.

Huang CF, Hsu CN, Chien SJ, Lin YJ, Huang LT, Tain YL
European journal of pharmacology 2013 Jan 15;699(1-3):233-40

Nebivolol does not protect against 5/6 ablation/infarction induced chronic kidney disease in rats - comparison with angiotensin II receptor blockade.

Sasser JM, Moningka NC, Tsarova T, Baylis C
Life sciences 2012 Jul 26;91(1-2):54-63

Endotoxemia exacerbates kidney injury and increases asymmetric dimethylarginine in young bile duct-ligated rats.

Huang LT, Hung JF, Chen CC, Hsieh CS, Yu HR, Hsu CN, Tain YL
Shock (Augusta, Ga.) 2012 Apr;37(4):441-8

Aliskiren prevents hypertension and reduces asymmetric dimethylarginine in young spontaneously hypertensive rats.

Tain YL, Hsu CN, Lin CY, Huang LT, Lau YT
European journal of pharmacology 2011 Nov 30;670(2-3):561-5

Renal nitric oxide production in rat pregnancy: role of constitutive nitric oxide synthases.

Smith CA, Santymire B, Erdely A, Venkat V, Losonczy G, Baylis C
American journal of physiology. Renal physiology 2010 Oct;299(4):F830-6

Melatonin prevents hypertension and increased asymmetric dimethylarginine in young spontaneous hypertensive rats.

Tain YL, Huang LT, Lin IC, Lau YT, Lin CY
Journal of pineal research 2010 Nov;49(4):390-8

Splice variants of neuronal nitric oxide synthase are present in the rat kidney.

Smith C, Merchant M, Fekete A, Nyugen HL, Oh P, Tain YL, Klein JB, Baylis C
Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association 2009 May;24(5):1422-8

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of nNOS was performed by loading 25 µg 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of nNOS was performed by loading 25 µg 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

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 2 Nitric oxide synthase (NOS)-1 and -3 are involvd in production of basal NO in MECs. ( A ) (Top) NOS-1, -2 or -3 staining of the MCF10A breast cancer progression series. Scale bars: 20 mum. (Bottom) Quantification of NOS signal/cell. AU: Arbitrary unit. ( B ) (Left) NOS-1, -2 or -3 staining of mouse mammary glands (10 weeks old, virgin female). Nuclei were counterstained with DAPI (blue). Scale bars: 50 mum. (Right) Quantification of NOS signal/cell. ( C ) (Left) S-NOC staining of MCF10A cells expressing scramble, NOS-1, -2 or -3 shRNA. Nuclei were counterstained with DAPI (blue). Scale bars: 20 mum. (Right) Quantification of SNOC signal/cell. ( D ) S-NOC staining of MCF10A cells with double knockdown (KD) of NOS-1 and -3 using different combinations of shRNA and siRNA. (Top 2 rows, left) S-NOC staining of MCF10A cells expressing scramble, NOS-2 or -1 shRNA, which were further transfected with NOS-3 siRNA. (Top right) Quantification of S-NOC signal/cell. (Bottom 2 rows, left) S-NOC staining of MCF10A cells expressing scramble, NOS-2 or -3 shRNA, which were further transfected with a NOS-1 siRNA. (Bottom right) Quantification of S-NOC signal/cell. Note that while KD of NOS-1 or -3 alone partially inhibited SNO, double KD of NOS-1/NOS-3 abrogated SNOC. Scale bars: 50 mum.

PA1-033

Antibody data