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General description
NF-kappa-B inhibitor alpha (UniProt: P25963; also known as I-kappa-B-alpha, IkB-alpha, IkappaB alpha, Major histocompatibility complex enhancer-binding protein MAD3) is encoded by the NFKBIA (also known as IKBA, MAD3, NFKBI) gene (Gene ID: 4792) in human. IkBa is known to inhibit the activity of dimeric NF-kB/REL complexes by trapping Rel dimers in the cytoplasm through masking of their nuclear localization signals. In most resting cells, NF-kB is sequestered in the cytoplasm in an inactive form associated with IkBa, which blocks the ability of NF-kB translocate to the nucleus. However, following exposure to pro-inflammatory cytokines, UV light, reactive oxygen species, or bacterial and viral toxins, IkBa is phosphorylated by IkB kinases (IKK) on serine 32 and 36 that allows its polyubiquitination and proteasomal degradation. This unmasks the nuclear localization signal of NF-kB and allows it to translocate to the nucleus where it binds to NF-kB response elements in target genes and regulates their transcription. In the nucleus, it can bind to NF-kB and facilitate its nuclear export. IkB can shuttle between the cytoplasm and nucleus through its nuclear import signal (aa 111-120) and nuclear export signal (aa 45-54). This ZooMAbZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals.
We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information.
ZooMAb® antibodies represent an entirely new generation of recombinant monoclonal antibodies.Each ZooMAb® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb® antibodies are reliably available and ready to ship when you need them.
Immunogen
KLH-conjugated linear peptide corresponding to 10 amino acids surrounding phosphoserine 32 from human IkBa.
Application
Quality Control Testing
Evaluated by Western Blotting in lysate from HeLa cells treated with TNF- .
Western Blotting Analysis (WB): A 1:1,000 dilution of this antibody detected IkB in lysate from overnight starved HeLa cells treated with TNF- (100 ng/mL), but not in untreated cells.
Tested applications
Western Blotting Analysis: A 1:1,000 dilution of this antibody detected a construct containing GST-tagged recombinant IkBa protein fragment phosphorylated on Ser32 but did not detect in non-phosphorylated construct.
Affinity Binding Assay: A representative lot of this antibody bound phospho-IkB -Ser32 with a KD of 6.9 x 10-7 in an affinity binding assay.
Immunocytochemistry Analysis: A 1:1,000 dilution from a representative lot detected phospho- IkB -Ser32 in HeLa cells.
Peptide Inhibition Assay: Target band detection in a construct containing GST-tagged recombinant IkBa protein fragment phosphorylated on Ser32 was prevented by pre-blocking of a representative lot with the immunogen phosphopeptide, but not the corresponding non-phosphopeptide.
Flow Cytometry Analysis: 0.1 μg from a representative lot detected phospho-IkB -Ser32 in one million HeLa cells.
Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user
Biochem/physiol Actions
Clone 11D19 is a ZooMAb® rabbit recombinant monoclonal antibody that specifically detects IkBa phosphorylated on serine 32.
Physical form
Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL.
Preparation Note
Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.
Reconstitute lyophilized antibody pellet with 25μL of ultrapure water or Phosphate Buffered Saline (PBS). Please refer to our reconstitution protocol and the specific application guidance on the suggested starting dilutions and sample type.
Legal Information
ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
| biological source | rabbit |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| conjugate | unconjugated |
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| clone | 11D19, recombinant monoclonal |
| description | recombinant, expressed in HEK 293 cells |
| product line | ZooMAb® learn more |
| form | lyophilized |
| mol wt | calculated mol wt 35.61 kDa,observed mol wt ~40 kDa |
| purified by | using Protein A |
| species reactivity | human |
| species reactivity (predicted by homology) | porcine, monkey, feline, canine, bovine |
| packaging | antibody small pack of 25 μL |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry. |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation |
| sustainability | Greener Alternative Product |
| technique(s) | affinity binding assay: suitable,flow cytometry: suitable,immunocytochemistry: suitable,inhibition assay: suitable,western blot: suitable |
| isotype | IgG |
| epitope sequence | N-terminus |
| Protein ID accession no. | NP_065390 |
| UniProt accession no. | P25963 |
| greener alternative category | , Aligned |
| shipped in | ambient |
| storage temp. | 2-8°C |
| target post-translational modification | phosphorylation (pSer32) |
| Gene Information | human ... NFKBIA(4792) |
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