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FCM (Flow Cytometry) (Figure 6. Flow Cytometry analysis of A549 cells using anti-NIT2 antibody (AAA19853).Overlay histogram showing A549 cells stained with AAA19853 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-NIT2 Antibody (AAA19853, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

Rabbit NIT2 Polyclonal Antibody | anti-NIT2 antibody

Anti-NIT2 Antibody Picoband

Gene Names
NIT2; HEL-S-8a
Reactivity
Human, Mouse, Rat
Applications
Western Blot, Immunohistochemistry, Flow Cytometry, Functional Assay, ELISA
Purity
Immunogen affinity purified.
Synonyms
NIT2; Polyclonal Antibody; Anti-NIT2 Antibody Picoband; troponin T3; fast skeletal type; Troponin T; fast skeletal muscle; TnTf; Beta-TnTF; Fast skeletal muscle troponin T; fTnT; TNNT3; anti-NIT2 antibody
Ordering
For Research Use Only!
Host
Rabbit
Reactivity
Human, Mouse, Rat
Clonality
Polyclonal
Isotype
Rabbit IgG
Specificity
In fetal and adult fast skeletal muscles, with a higher level expression in fetal than in adult muscle.
Purity/Purification
Immunogen affinity purified.
Form/Format
Lyophilized. Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Applicable Applications for anti-NIT2 antibody
Western Blot (WB), Immunohistochemistry (IHC), Flow Cytometry (FC/FACS), ELISA (EIA)
Application Notes
WB: 0.25-0.5ug/ml, Human, Rat
IHC(Paraffin-embedded Section): 2-5ug/ml, Human, Mouse, Rat
FC/FACS (Fixed): 1-3ug/1x106 cells, Human
ELISA: 0.1-0.5ug/ml
Immunogen
E coli-derived human NIT2 recombinant protein (Position: M1-K249).
Subcellular Localization
Troponin complex, Cytosol
Reconstitution
Adding 0.2 ml of distilled water will yield a concentration of 500ug/ml.
Cross Reactivity
No cross-reactivity with other proteins.
Protein Function
Troponin T is the tropomyosin-binding subunit of troponin, the thin filament regulatory complex which confers calcium-sensitivity to striated muscle actomyosin ATPase activity.
Preparation and Storage
At -20 degree C for one year from date of receipt. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for six months.Avoid repeated freezing and thawing.

FCM (Flow Cytometry)

(Figure 6. Flow Cytometry analysis of A549 cells using anti-NIT2 antibody (AAA19853).Overlay histogram showing A549 cells stained with AAA19853 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-NIT2 Antibody (AAA19853, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

FCM (Flow Cytometry) (Figure 6. Flow Cytometry analysis of A549 cells using anti-NIT2 antibody (AAA19853).Overlay histogram showing A549 cells stained with AAA19853 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-NIT2 Antibody (AAA19853, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

IHC (Immunohistochemistry)

(Figure 5. IHC analysis of NIT2 using anti-NIT2 antibody (AAA19853).NIT2 was detected in a paraffin-embedded section of rat kidney tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-NIT2 Antibody (AAA19853) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-NIT2 Antibody (AAA19853) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-NIT2 Antibody (AAA19853) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-NIT2 Antibody (AAA19853) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Lane 2: rat liver tissue lysates,Lane 3: rat kidney tissue lysates.After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-NIT2 antigen affinity purified polyclonal antibody (#AAA19853) at 0.5ug/mL overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for NIT2 at approximately 33 kDa. The expected band size for NIT2 is at 33 kDa.)

IHC (Immunohistochemistry) (Figure 5. IHC analysis of NIT2 using anti-NIT2 antibody (AAA19853).NIT2 was detected in a paraffin-embedded section of rat kidney tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-NIT2 Antibody (AAA19853) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-NIT2 Antibody (AAA19853) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-NIT2 Antibody (AAA19853) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-NIT2 Antibody (AAA19853) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Lane 2: rat liver tissue lysates,Lane 3: rat kidney tissue lysates.After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-NIT2 antigen affinity purified polyclonal antibody (#AAA19853) at 0.5ug/mL overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for NIT2 at approximately 33 kDa. The expected band size for NIT2 is at 33 kDa.)
Related Product Information for anti-NIT2 antibody
The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
References
1. Asakura, M., Nakano, M., Hayashida, K., Fujii, H., Nakajima, M., Atsuda, K., Itoh, T., Fujiwara, R. Human nitrilase-like protein does not catalyze the hydrolysis of vildagliptin. Drug Metab. Pharmacokinet. 29: 463-469, 2014. 2. Chien, C.-H., Gao, Q.-Z., Cooper, A. J. L., Lyu, J.-H., Sheu, S.-Y. Structural insights into the catalytic active site and activity of human Nit2/omega-amidase: kinetic assay and molecular dynamics simulation. J. Biol. Chem. 287: 25715-25726, 2012. 3. Jaisson, S., Veiga-da-Cunha, M., Van Schaftingen, E. Molecular identification of omega-amidase, the enzyme that is functionally coupled with glutamine transaminases, as the putative tumor suppressor Nit2. Biochimie 91: 1066-1071, 2009.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
UniProt Accession #
Molecular Weight
276
NCBI Official Full Name
Omega-amidase NIT2
NCBI Official Synonym Full Names
nitrilase family, member 2
NCBI Official Symbol
NIT2
NCBI Official Synonym Symbols
HEL-S-8a
NCBI Protein Information
omega-amidase NIT2; Nit protein 2; nitrilase homolog 2; epididymis secretory sperm binding protein Li 8a
UniProt Protein Name
Omega-amidase NIT2
UniProt Gene Name
NIT2
UniProt Entry Name
NIT2_HUMAN

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Product Notes

The NIT2 nit2 (Catalog #AAA19853) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-NIT2 Antibody Picoband reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's NIT2 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry (IHC), Flow Cytometry (FC/FACS), ELISA (EIA). WB: 0.25-0.5ug/ml, Human, Rat IHC(Paraffin-embedded Section): 2-5ug/ml, Human, Mouse, Rat FC/FACS (Fixed): 1-3ug/1x106 cells, Human ELISA: 0.1-0.5ug/ml. Researchers should empirically determine the suitability of the NIT2 nit2 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "NIT2, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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