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Product Info Summary
| SKU: | A00738-2 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-Nkx2.5/NKX2-5 Antibody Picoband®
SKU/Catalog Number
A00738-2
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Nkx2.5/NKX2-5 Antibody Picoband® catalog # A00738-2. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human. 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.
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-Nkx2.5/NKX2-5 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00738-2)
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 Nkx2.5/NKX2-5 recombinant protein (Position: K15-R132).
*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
A00738-2 is reactive to NKX2-5 in Human
Reconstitution
Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
Observed Molecular Weight
35 kDa
Calculated molecular weight
34.918kDa
Background of NKX2.5
Homeobox protein Nkx-2.5, also known as NKX2E or CSX is a protein that in humans is encoded by the NKX2-5 gene. It is mapped to 5q35.1. Homeobox-containing genes play critical roles in regulating tissue-specific gene expression essential for tissue differentiation, as well as determining the temporal and spatial patterns of development. Nkx2.5 and Tbx5 directly bound to the promoter of the gene encoding cardiac-specific natriuretic peptide precursor type A (NPPA) in tandem, and both transcription factors showed synergistic activation. The cardiac homeobox protein Nkx2.5 is essential in cardiac development, and mutations in CSX (which encodes Nkx2.5) cause various congenital heart malformations.
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.
Application & Images
Applications
A00738-2 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee
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
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human
ELISA, 0.1-0.5 μg/ml, -
Positive Control
WB: human U20S whole cell, human U251 whole cell, human SiHa whole cell
ICC/IF: SiHa cell
FCM: 293T cell
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of Nkx2.5/NKX2-5 using anti-Nkx2.5/NKX2-5 antibody (A00738-2).
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 U20S whole cell lysates,
Lane 2: human U251 whole cell lysates,
Lane 3: human SiHa whole cell 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-Nkx2.5/NKX2-5 antigen affinity purified polyclonal antibody (Catalog # A00738-2) 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 Nkx2.5/NKX2-5 at approximately 35 kDa. The expected band size for Nkx2.5/NKX2-5 is at 35 kDa.
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Figure 2. IF analysis of Nkx2.5/NKX2-5 using anti-Nkx2.5/NKX2-5 antibody (A00738-2).
Nkx2.5/NKX2-5 was detected in an immunocytochemical section of SiHa cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-Nkx2.5/NKX2-5 Antibody (A00738-2) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
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Figure 3. Flow Cytometry analysis of 293T cells using anti-Nkx2.5/NKX2-5 antibody (A00738-2).
Overlay histogram showing 293T cells stained with A00738-2 (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-Nkx2.5/NKX2-5 Antibody (A00738-2, 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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Protein Target Info & Infographic
Gene/Protein Information For NKX2-5 (Source: Uniprot.org, NCBI)
Gene Name
NKX2-5
Full Name
Homeobox protein Nkx-2.5
Weight
34.918kDa
Superfamily
NK-2 homeobox family
Alternative Names
Interstitial collagenase; Fibroblast collagenase; Matrix metalloproteinase-1; MMP-1; 22 kDa interstitial collagenase; 27 kDa interstitial collagenase; MMP1; CLG NKX2-5 CHNG5, CSX, CSX1, HLHS2, NKX2.5, NKX2E, NKX4-1, VSD3 NK2 homeobox 5 homeobox protein Nkx-2.5|NK2 transcription factor related, locus 5|NKX 2-5|cardiac-specific homeobox 1|homeobox protein CSX|homeobox protein NK-2 homolog E|homeobox protein NKX 2-5|tinman homolog|tinman paralog
*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 NKX2-5, check out the NKX2-5 Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for NKX2-5: 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-Nkx2.5/NKX2-5 Antibody Picoband® (A00738-2)
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