Product Info Summary
SKU: | A05891-3 |
---|---|
Size: | 100 µg/vial |
Reactive Species: | Human, Mouse, Rat |
Host: | Rabbit |
Application: | ELISA, Flow Cytometry, IHC, WB |
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Product info
Product Name
Anti-NIT2 Antibody Picoband™
SKU/Catalog Number
A05891-3
Size
100 µg/vial
Form
Lyophilized
Description
Boster Bio Anti-NIT2 Antibody Picoband™ catalog # A05891-3. Tested in ELISA, IHC, WB, Flow Cytometry applications. This antibody reacts with Human, Mouse, Rat.
Storage & Handling
At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.
Cite This Product
Anti-NIT2 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A05891-3)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human NIT2 recombinant protein (Position: M1-K249).
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A05891-3 is reactive to Nit2 in Human, Mouse, Rat
Applications
A05891-3 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee
Observed Molecular Weight
33 kDa
Calculated molecular weight
Background of Nit2
NIT2 belongs to a branch of the nitrilase superfamily of enzymes that cleave carbon-nitrogen bonds. NIT2 functions as an omega-amidase and catalyzes hydrolysis of alpha-ketoglutaramate, forming alpha-ketoglutarate and ammonia. This reaction is also functionally coupled with a subset of transaminases that reaminate the keto acid analogs of some essential amino acids, most particularly methionine and phenylalanine.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Assay dilution & Images
Reconstitution
Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.25-0.5 µg/ml, Human, Rat
Immunohistochemistry(Paraffin-embedded Section), 2-5 µg/ml, Human, Mouse, Rat
Flow Cytometry (Fixed), 1-3 µg/1x106 cells, Human
Direct ELISA, 0.1-0.5 µg/ml, Human
Validation Images & Assay Conditions
![a05891 3 nit2 primary antibodies wb testing 1 a05891 3 nit2 primary antibodies wb testing 1](https://www.bosterbio.com/media/catalog/product/a/0/a05891-3-nit2-primary-antibodies-wb-testing-1.jpg)
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Figure 1. Western blot analysis of NIT2 using anti-NIT2 antibody (A05891-3).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human A549 whole cell lysates,
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 (Catalog # A05891-3) at 0.5 μg/mL overnight at 4°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 (Catalog # EK1002) 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.
![a05891 3 nit2 primary antibodies ihc testing 2 a05891 3 nit2 primary antibodies ihc testing 2](https://www.bosterbio.com/media/catalog/product/a/0/a05891-3-nit2-primary-antibodies-ihc-testing-2.jpg)
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Figure 2. IHC analysis of NIT2 using anti-NIT2 antibody (A05891-3).
NIT2 was detected in a paraffin-embedded section of human squamous cell carcinoma penis 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 2 μg/ml rabbit anti-NIT2 Antibody (A05891-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
![a05891 3 nit2 primary antibodies ihc testing 3 a05891 3 nit2 primary antibodies ihc testing 3](https://www.bosterbio.com/media/catalog/product/a/0/a05891-3-nit2-primary-antibodies-ihc-testing-3.jpg)
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Figure 3. IHC analysis of NIT2 using anti-NIT2 antibody (A05891-3).
NIT2 was detected in a paraffin-embedded section of human liver cancer 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 2 μg/ml rabbit anti-NIT2 Antibody (A05891-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
![a05891 3 nit2 primary antibodies ihc testing 4 a05891 3 nit2 primary antibodies ihc testing 4](https://www.bosterbio.com/media/catalog/product/a/0/a05891-3-nit2-primary-antibodies-ihc-testing-4.jpg)
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Figure 4. IHC analysis of NIT2 using anti-NIT2 antibody (A05891-3).
NIT2 was detected in a paraffin-embedded section of mouse 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 2 μg/ml rabbit anti-NIT2 Antibody (A05891-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
![a05891 3 nit2 primary antibodies ihc testing 5 a05891 3 nit2 primary antibodies ihc testing 5](https://www.bosterbio.com/media/catalog/product/a/0/a05891-3-nit2-primary-antibodies-ihc-testing-5.jpg)
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Figure 5. IHC analysis of NIT2 using anti-NIT2 antibody (A05891-3).
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 2 μg/ml rabbit anti-NIT2 Antibody (A05891-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
![a05891 3 nit2 primary antibodies fcm testing 6 a05891 3 nit2 primary antibodies fcm testing 6](https://www.bosterbio.com/media/catalog/product/a/0/a05891-3-nit2-primary-antibodies-fcm-testing-6.png)
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Figure 6. Flow Cytometry analysis of A549 cells using anti-NIT2 antibody (A05891-3).
Overlay histogram showing A549 cells stained with A05891-3 (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 (A05891-3, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Protein Target Info & Infographic
Gene/Protein Information For Nit2 (Source: Uniprot.org, NCBI)
Gene Name
Nit2
Full Name
Omega-amidase NIT2
Weight
Superfamily
carbon-nitrogen hydrolase superfamily
Alternative Names
Omega-amidase NIT2
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on Nit2, check out the Nit2 Infographic
![Nit2 infographic](/media/images/gene-infographic-example.jpg)
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for Nit2: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-NIT2 Antibody Picoband™ (A05891-3)
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