Development of an Analytical Assay for Electrochemical Detection and Quantification of Protein-Bound 3-Nitrotyrosine in Biological Samples and Comparison with Classical, Antibody-Based Methods.

Development of an Analytical Assay for Electrochemical Detection and Quantification of Protein-Bound 3-Nitrotyrosine in Biological Samples and Comparison with Classical, Antibody-Based Methods.

Reactive oxygen and nitrogen species (RONS) trigger oxidative harm, which is related to endothelial dysfunction and heart problems, however may contribute to redox signaling. Due to this fact, their exact detection is necessary for the analysis of illness mechanisms.
Right here, we in contrast three totally different strategies for the detection of 3-nitrotyrosine (3-NT), a marker of nitro-oxidative stress, in organic samples. Nitrated proteins had been generated by incubation with peroxynitrite or 3-morpholino sydnonimine (Sin-1) and subjected to whole hydrolysis utilizing pronase, a mix of various proteases.
The three-NT was then separated by excessive efficiency liquid chromatography (HPLC) and quantified by electrochemical detection (ECD, CoulArray) and in comparison with classical strategies, particularly enzyme-linked immunosorbent assay (ELISA) and dot blot evaluation utilizing particular 3-NT antibodies.
Calibration curves for genuine 3-NT (detection restrict 10 nM) and a concentration-response sample for 3-NT obtained from digested nitrated bovine serum albumin (BSA) had been extremely linear over a large 3-NT focus vary.
Additionally, ex vivo nitration of protein from coronary heart, remoted mitochondria, and serum/plasma could possibly be quantified utilizing the HPLC/ECD technique and was confirmed by LC-MS/MS. Of word, nitro-oxidative harm of mitochondria ends in elevated superoxide (O2•-) formation charges (measured by dihydroethidium-based HPLC assay), pointing to a self-amplification mechanism of oxidative stress.
Primarily based on our ex vivo knowledge, the CoulArray quantification technique for 3-NT appears to have some benefits concerning sensitivity and selectivity. Establishing a dependable automated HPLC assay for the routine quantification of 3-NT in organic samples of cell tradition, of animal and human origin appears to be extra subtle than anticipated.

Anti-nitrotyrosine antibodies for immunohistochemistry.

Nitrotyrosine is a crucial marker for the formation of peroxynitrite and probably different reactive nitrogen species derived from nitric oxide in vivo (1). Pathological circumstances can considerably improve the manufacturing of nitric oxide, but this molecule itself doesn’t usually yield nitration of tyrosine residues in proteins when added to organic samples (1,2).
Nonetheless nitric oxide reacts at close to diffusion-limited charges with superoxide (O(2) (-)) to type the robust oxidant peroxynitrite (ONOO(-)) (3). Nitration on the 3-position of tyrosine is a serious product of peroxynitrite assault on proteins (4,5).
Definitely, small quantities of nitrotyrosine will be produced in vivo by different mechanisms (6), however peroxynitrite is by far essentially the most environment friendly mechanism for nitrating tyrosine below biologically related circumstances with pure antioxidants and different targets current.

Era of novel recombinant antibodies towards nitrotyrosine by antibody phage show.

Nitrotyrosine is a posttranslational protein modification that happens below oxidative and nitrosative stress, and performs an necessary position in quite a few pathological circumstances. To analyse nitrotyrosine formation a number of industrial monoclonal and polyclonal antibodies reacting with 3-nitrotyrosine have been developed which nevertheless don’t work correctly in all required assays.
Right here, antibody phage show was used to pick out recombinant antibodies that particularly react with nitrotyrosine in varied protein contexts. 9 preliminary alternatives had been carried out, utilizing artificial peptides, peroxynitrite-modified proteins and conjugated proteins as antigens. 4 antibodies had been remoted that every exhibited a attribute binding reactivity that drastically relied on the antigens that had been used for his or her alternatives.
Generally, the alternatives utilizing small, artificial and biotinylated peptides had been essentially the most profitable method. Subsequently, antibody 11B1 was affinity matured by error inclined mutagenesis, ensuing within the isolation of two antibodies, designated 47A7 and 47B1.
Competitors ELISA and immunoblotting after therapy with sodium dithionite additional demonstrated the specificity of antibody 47B1 for nitrotyrosine. The outcomes introduced right here display that antibody phage show is a helpful technique to isolate antibodies towards posttranslational modifications, that are highly effective instruments within the proteomic period.

Epitope motif of an anti-nitrotyrosine antibody particular for tyrosine-nitrated peptides revealed by a mix of affinity approaches and mass spectrometry.

Nitration of tyrosine residues has been proven to be an necessary oxidative modification in proteins and has been advised to play a task in a number of ailments equivalent to atherosclerosis, bronchial asthma, lung and neurodegenerative ailments.
Detection of nitrated proteins has been primarily based mostly on the usage of nitrotyrosine-specific antibodies. In distinction, solely a small variety of nitration websites in proteins have been unequivocally recognized by MS. We have now used a monoclonal 3-NT-specific antibody, and have synthesized a sequence of tyrosine-nitrated peptides of prostacyclin synthase (PCS) by which a single particular nitration web site at Tyr-430 had been beforehand recognized upon response with peroxynitrite17.
The willpower of antibody-binding affinity and specificity of PCS peptides nitrated at totally different tyrosine residues (Tyr-430, Tyr-421, Tyr-83) and sequence mutations across the nitration websites offered the identification of an epitope motif containing positively charged amino acids (Lys and/or Arg) N-terminal to the nitration web site.
The best affinity to the anti-3NT-antibody was discovered for the PCS peptide comprising the Tyr-430 nitration web site with a Okay(D) of 60 nM decided for the peptide, PCS(424-436-Tyr-430NO(2) ); in distinction, PCS peptides nitrated at Tyr-421 and Tyr-83 had considerably decrease affinity.
ELISA, SAW bioaffinity, proteolytic digestion of antibody-bound peptides and affinity-MS evaluation revealed highest affinity to the antibody for tyrosine-nitrated peptides that contained positively charged amino acids within the N-terminal sequence to the nitration web site.
Remarkably, related N-terminal sequences of tyrosine-nitration websites have been not too long ago recognized in nitrated physiological proteins, equivalent to eosinophil peroxidase and eosinophil-cationic protein.

Antibodies towards nitric oxide broken poly L-tyrosine and 3-nitrotyrosine ranges in systemic lupus erythematosus.

Alterations within the amino acid construction or sequence can generate neo-epitopes from self-proteins inflicting autoaggressive immune assault.
Reactive nitrogen species are an necessary issue that induces post-translational modification of proteins by mobile discount and oxidation mechanism; cysteinyl-nitrosylation or tyrosine nitration resulting in doubtlessly pathogenic pathways. It was considered curiosity to analyze the immunogenicity of nitrated poly L-tyrosine vis-á-vis its doable position within the induction of antibodies in systemic lupus erythematosus (SLE).
Commercially out there poly L-tyrosine was uncovered to nitrating species and the harm was monitored by UV spectroscopy and alkaline gel electrophoresis. The outcomes indicated the formation of 3-nitrotyrosine. Nitrated poly L-tyrosine induced greater titre antibodies as in comparison with the native type.

Nitrotyrosine Antibody

abx140246-01mg 0.1 mg
EUR 537
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Nitrotyrosine Antibody

abx440039-100ug 100 ug
EUR 592
  • Shipped within 5-12 working days.

Nitrotyrosine Antibody

abx440320-100ug 100 ug
EUR 592
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Nitrotyrosine Antibody

abx440601-100ug 100 ug
EUR 592
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Nitrotyrosine Antibody

abx440882-100ug 100 ug
EUR 592
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Nitrotyrosine Antibody

abx441163-100ug 100 ug
EUR 592
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Nitrotyrosine Antibody

abx441444-100ug 100 ug
EUR 592
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Nitrotyrosine Antibody

abx441725-100ug 100 ug
EUR 592
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Nitrotyrosine Antibody

abx442006-100ug 100 ug
EUR 592
  • Shipped within 5-12 working days.

Nitrotyrosine Antibody

abx442287-100ug 100 ug
EUR 592
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Nitrotyrosine Antibody

abx442568-100ug 100 ug
EUR 592
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Nitrotyrosine Antibody

abx442849-100ug 100 ug
EUR 592
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Nitrotyrosine Antibody

abx443129-100ug 100 ug
EUR 578
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Nitrotyrosine Antibody

abx443409-100ug 100 ug
EUR 578
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Nitrotyrosine Antibody

abx443690-100ug 100 ug
EUR 606
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Nitrotyrosine Antibody

abx443971-100ug 100 ug
EUR 592
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Nitrotyrosine Antibody

abx444252-100ug 100 ug
EUR 592
  • Shipped within 5-12 working days.

Nitrotyrosine Antibody

abx444533-100ug 100 ug
EUR 592
  • Shipped within 5-12 working days.

Nitrotyrosine Antibody

20-abx444883
  • EUR 537.00
  • EUR 342.00
  • 100 ug
  • 25 ug
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Anti-Nitrotyrosine Antibody

A1300-50
EUR 425

Nitrotyrosine Antibody (7A5)

5413-50
EUR 327

Nitrotyrosine Antibody (HM11)

5414-50
EUR 648

Nitrotyrosine (NT) Antibody

20-abx100841
  • EUR 411.00
  • EUR 133.00
  • EUR 1135.00
  • EUR 551.00
  • EUR 314.00
  • 100 ug
  • 10 ug
  • 1 mg
  • 200 ug
  • 50 ug
  • Shipped within 5-7 working days.

Anti-Nitrotyrosine antibody

STJ16100894 1 mL
EUR 749

Anti-Nitrotyrosine antibody

STJ16100895 0.5 ml
EUR 611

Nitrotyrosine (BSA)

20-abx165786
  • EUR 578.00
  • EUR 258.00
  • EUR 1720.00
  • EUR 690.00
  • EUR 425.00
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  • 10 ug
  • 1 mg
  • 200 ug
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anti-Nitrotyrosine

LF-PA0067 100 ul
EUR 334
Description: Rabbit polyclonal to Nitrotyrosine

Nitrotyrosine Antibody (NOY-7A5)

5415-50
EUR 501

Monoclonal antibody for Nitrotyrosine

SMC-154C 0.025mg
EUR 211
  • Protein tyrosine nitration results in a post-translational modification that is increasingly receiving attention as an important component of nitric oxide signaling (2). While multiple nonenzymatic mechanisms are known to be capable of producing nitr
  • Show more
Description: A monoclonal antibody from clone 39B6 against Human | Mouse | Rat | Bovine Nitrotyrosine. The host species for the production of this antibody is Mouse. The antigen used for immunization is 3-(4-hydroxy-3-nitrophenylacetamido) propionic acid-bovine serum albumin. The antibody is tested and validated for WB, IHC, ICC/IF, IP, ELISA, FCM, AM assays with the following recommended dilutions: WB (1:1400), IHC (1:100). This MAb for Nitrotyrosine is not conjugated.

Monoclonal antibody for Nitrotyrosine

SMC-154D 0.1mg
EUR 364
  • Protein tyrosine nitration results in a post-translational modification that is increasingly receiving attention as an important component of nitric oxide signaling (2). While multiple nonenzymatic mechanisms are known to be capable of producing nitr
  • Show more
Description: A monoclonal antibody from clone 39B6 against Human | Mouse | Rat | Bovine Nitrotyrosine. The host species for the production of this antibody is Mouse. The antigen used for immunization is 3-(4-hydroxy-3-nitrophenylacetamido) propionic acid-bovine serum albumin. The antibody is tested and validated for WB, IHC, ICC/IF, IP, ELISA, FCM, AM assays with the following recommended dilutions: WB (1:1400), IHC (1:100). This MAb for Nitrotyrosine is not conjugated.

Monoclonal antibody for Nitrotyrosine

SMC-154D-A390 0.1mg
EUR 411
  • Protein tyrosine nitration results in a post-translational modification that is increasingly receiving attention as an important component of nitric oxide signaling (2). While multiple nonenzymatic mechanisms are known to be capable of producing nitr
  • Show more
Description: A monoclonal antibody from clone 39B6 against Human | Mouse | Rat | Bovine Nitrotyrosine. The host species for the production of this antibody is Mouse. The antigen used for immunization is 3-(4-hydroxy-3-nitrophenylacetamido) propionic acid-bovine serum albumin. The antibody is tested and validated for WB, IHC, ICC/IF, IP, ELISA, FCM, AM assays with the following recommended dilutions: WB (1:1400), IHC (1:100). This MAb for Nitrotyrosine is conjugated with ATTO 390.

Monoclonal antibody for Nitrotyrosine

SMC-154D-A488 0.1mg
EUR 410
  • Protein tyrosine nitration results in a post-translational modification that is increasingly receiving attention as an important component of nitric oxide signaling (2). While multiple nonenzymatic mechanisms are known to be capable of producing nitr
  • Show more
Description: A monoclonal antibody from clone 39B6 against Human | Mouse | Rat | Bovine Nitrotyrosine. The host species for the production of this antibody is Mouse. The antigen used for immunization is 3-(4-hydroxy-3-nitrophenylacetamido) propionic acid-bovine serum albumin. The antibody is tested and validated for WB, IHC, ICC/IF, IP, ELISA, FCM, AM assays with the following recommended dilutions: WB (1:1400), IHC (1:100). This MAb for Nitrotyrosine is conjugated with ATTO 488.

Monoclonal antibody for Nitrotyrosine

SMC-154D-A565 0.1mg
EUR 410
  • Protein tyrosine nitration results in a post-translational modification that is increasingly receiving attention as an important component of nitric oxide signaling (2). While multiple nonenzymatic mechanisms are known to be capable of producing nitr
  • Show more
Description: A monoclonal antibody from clone 39B6 against Human | Mouse | Rat | Bovine | Monkey | Dog | Rabbit Nitrotyrosine. The host species for the production of this antibody is Mouse. The antigen used for immunization is 3-(4-hydroxy-3-nitrophenylacetamido) propionic acid-bovine serum albumin. The antibody is tested and validated for WB, IHC, ICC/IF, IP, ELISA, FCM, AM assays with the following recommended dilutions: WB (1:1400), IHC (1:100). This MAb for Nitrotyrosine is conjugated with ATTO 565.

Monoclonal antibody for Nitrotyrosine

SMC-154D-A594 0.1mg
EUR 410
  • Protein tyrosine nitration results in a post-translational modification that is increasingly receiving attention as an important component of nitric oxide signaling (2). While multiple nonenzymatic mechanisms are known to be capable of producing nitr
  • Show more
Description: A monoclonal antibody from clone 39B6 against Human | Mouse | Rat | Bovine | Monkey | Dog | Rabbit Nitrotyrosine. The host species for the production of this antibody is Mouse. The antigen used for immunization is 3-(4-hydroxy-3-nitrophenylacetamido) propionic acid-bovine serum albumin. The antibody is tested and validated for WB, IHC, ICC/IF, IP, ELISA, FCM, AM assays with the following recommended dilutions: WB (1:1400), IHC (1:100). This MAb for Nitrotyrosine is conjugated with ATTO 594.

Monoclonal antibody for Nitrotyrosine

SMC-154D-A633 0.1mg
EUR 410
  • Protein tyrosine nitration results in a post-translational modification that is increasingly receiving attention as an important component of nitric oxide signaling (2). While multiple nonenzymatic mechanisms are known to be capable of producing nitr
  • Show more
Description: A monoclonal antibody from clone 39B6 against Human | Mouse | Rat | Bovine | Monkey | Dog | Rabbit Nitrotyrosine. The host species for the production of this antibody is Mouse. The antigen used for immunization is 3-(4-hydroxy-3-nitrophenylacetamido) propionic acid-bovine serum albumin. The antibody is tested and validated for WB, IHC, ICC/IF, IP, ELISA, FCM, AM assays with the following recommended dilutions: WB (1:1400), IHC (1:100). This MAb for Nitrotyrosine is conjugated with ATTO 633.

Monoclonal antibody for Nitrotyrosine

SMC-154D-A655 0.1mg
EUR 410
  • Protein tyrosine nitration results in a post-translational modification that is increasingly receiving attention as an important component of nitric oxide signaling (2). While multiple nonenzymatic mechanisms are known to be capable of producing nitr
  • Show more
Description: A monoclonal antibody from clone 39B6 against Human | Mouse | Rat | Bovine | Monkey | Dog | Rabbit Nitrotyrosine. The host species for the production of this antibody is Mouse. The antigen used for immunization is 3-(4-hydroxy-3-nitrophenylacetamido) propionic acid-bovine serum albumin. The antibody is tested and validated for WB, IHC, ICC/IF, IP, ELISA, FCM, AM assays with the following recommended dilutions: WB (1:1400), IHC (1:100). This MAb for Nitrotyrosine is conjugated with ATTO 655.

Monoclonal antibody for Nitrotyrosine

SMC-154D-A680 0.1mg
EUR 410
  • Protein tyrosine nitration results in a post-translational modification that is increasingly receiving attention as an important component of nitric oxide signaling (2). While multiple nonenzymatic mechanisms are known to be capable of producing nitr
  • Show more
Description: A monoclonal antibody from clone 39B6 against Human | Mouse | Rat | Bovine | Monkey | Dog | Rabbit Nitrotyrosine. The host species for the production of this antibody is Mouse. The antigen used for immunization is 3-(4-hydroxy-3-nitrophenylacetamido) propionic acid-bovine serum albumin. The antibody is tested and validated for WB, IHC, ICC/IF, IP, ELISA, FCM, AM assays with the following recommended dilutions: WB (1:1400), IHC (1:100). This MAb for Nitrotyrosine is conjugated with ATTO 680.

Monoclonal antibody for Nitrotyrosine

SMC-154D-A700 0.1mg
EUR 410
  • Protein tyrosine nitration results in a post-translational modification that is increasingly receiving attention as an important component of nitric oxide signaling (2). While multiple nonenzymatic mechanisms are known to be capable of producing nitr
  • Show more
Description: A monoclonal antibody from clone 39B6 against Human | Mouse | Rat | Bovine | Monkey | Dog | Rabbit Nitrotyrosine. The host species for the production of this antibody is Mouse. The antigen used for immunization is 3-(4-hydroxy-3-nitrophenylacetamido) propionic acid-bovine serum albumin. The antibody is tested and validated for WB, IHC, ICC/IF, IP, ELISA, FCM, AM assays with the following recommended dilutions: WB (1:1400), IHC (1:100). This MAb for Nitrotyrosine is conjugated with ATTO 700.

Monoclonal antibody for Nitrotyrosine

SMC-154D-ALP 0.1mg
EUR 405
  • Protein tyrosine nitration results in a post-translational modification that is increasingly receiving attention as an important component of nitric oxide signaling (2). While multiple nonenzymatic mechanisms are known to be capable of producing nitr
  • Show more
Description: A monoclonal antibody from clone 39B6 against Human | Mouse | Rat | Bovine | Monkey | Dog | Rabbit Nitrotyrosine. The host species for the production of this antibody is Mouse. The antigen used for immunization is 3-(4-hydroxy-3-nitrophenylacetamido) propionic acid-bovine serum albumin. The antibody is tested and validated for WB, IHC, ICC/IF, IP, ELISA, FCM, AM assays with the following recommended dilutions: WB (1:1400), IHC (1:100). This MAb for Nitrotyrosine is conjugated with Alkaline Phosphatase.

Monoclonal antibody for Nitrotyrosine

SMC-154D-APC 0.1mg
EUR 409
  • Protein tyrosine nitration results in a post-translational modification that is increasingly receiving attention as an important component of nitric oxide signaling (2). While multiple nonenzymatic mechanisms are known to be capable of producing nitr
  • Show more
Description: A monoclonal antibody from clone 39B6 against Human | Mouse | Rat | Bovine | Monkey | Dog | Rabbit Nitrotyrosine. The host species for the production of this antibody is Mouse. The antigen used for immunization is 3-(4-hydroxy-3-nitrophenylacetamido) propionic acid-bovine serum albumin. The antibody is tested and validated for WB, IHC, ICC/IF, IP, ELISA, FCM, AM assays with the following recommended dilutions: WB (1:1400), IHC (1:100). This MAb for Nitrotyrosine is conjugated with APC.

Monoclonal antibody for Nitrotyrosine

SMC-154D-APCCY7 0.1mg
EUR 482
  • Protein tyrosine nitration results in a post-translational modification that is increasingly receiving attention as an important component of nitric oxide signaling (2). While multiple nonenzymatic mechanisms are known to be capable of producing nitr
  • Show more
Description: A monoclonal antibody from clone 39B6 against Human | Mouse | Rat | Bovine | Monkey | Dog | Rabbit Nitrotyrosine. The host species for the production of this antibody is Mouse. The antigen used for immunization is 3-(4-hydroxy-3-nitrophenylacetamido) propionic acid-bovine serum albumin. The antibody is tested and validated for WB, IHC, ICC/IF, IP, ELISA, FCM, AM assays with the following recommended dilutions: WB (1:1400), IHC (1:100). This MAb for Nitrotyrosine is conjugated with APC/Cy7.

Monoclonal antibody for Nitrotyrosine

SMC-154D-BI 0.1mg
EUR 407
  • Protein tyrosine nitration results in a post-translational modification that is increasingly receiving attention as an important component of nitric oxide signaling (2). While multiple nonenzymatic mechanisms are known to be capable of producing nitr
  • Show more
Description: A monoclonal antibody from clone 39B6 against Human | Mouse | Rat | Bovine | Monkey | Dog | Rabbit Nitrotyrosine. The host species for the production of this antibody is Mouse. The antigen used for immunization is 3-(4-hydroxy-3-nitrophenylacetamido) propionic acid-bovine serum albumin. The antibody is tested and validated for WB, IHC, ICC/IF, IP, ELISA, FCM, AM assays with the following recommended dilutions: WB (1:1400), IHC (1:100). This MAb for Nitrotyrosine is conjugated with Biotin.

Monoclonal antibody for Nitrotyrosine

SMC-154D-DY350 0.1mg
EUR 425
  • Protein tyrosine nitration results in a post-translational modification that is increasingly receiving attention as an important component of nitric oxide signaling (2). While multiple nonenzymatic mechanisms are known to be capable of producing nitr
  • Show more
Description: A monoclonal antibody from clone 39B6 against Human | Mouse | Rat | Bovine | Monkey | Dog | Rabbit Nitrotyrosine. The host species for the production of this antibody is Mouse. The antigen used for immunization is 3-(4-hydroxy-3-nitrophenylacetamido) propionic acid-bovine serum albumin. The antibody is tested and validated for WB, IHC, ICC/IF, IP, ELISA, FCM, AM assays with the following recommended dilutions: WB (1:1400), IHC (1:100). This MAb for Nitrotyrosine is conjugated with Dylight 350.

Monoclonal antibody for Nitrotyrosine

SMC-154D-DY405 0.1mg
EUR 413
  • Protein tyrosine nitration results in a post-translational modification that is increasingly receiving attention as an important component of nitric oxide signaling (2). While multiple nonenzymatic mechanisms are known to be capable of producing nitr
  • Show more
Description: A monoclonal antibody from clone 39B6 against Human | Mouse | Rat | Bovine | Monkey | Dog | Rabbit Nitrotyrosine. The host species for the production of this antibody is Mouse. The antigen used for immunization is 3-(4-hydroxy-3-nitrophenylacetamido) propionic acid-bovine serum albumin. The antibody is tested and validated for WB, IHC, ICC/IF, IP, ELISA, FCM, AM assays with the following recommended dilutions: WB (1:1400), IHC (1:100). This MAb for Nitrotyrosine is conjugated with Dylight 405.
Nitrated poly L-tyrosine was acknowledged by the autoantibodies current within the sera of sufferers affected by SLE by enzyme immunoassays and band shift assay. The doable position of nitrated self-proteins has been mentioned within the manufacturing of circulating anti-DNA antibodies in SLE.

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