Anti-Phospho-IκBα-Ser32/36 Antibody, clone 1I11 ZooMAb^® Rabbit Monoclonal

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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.

Clone 1I11 is a ZooMAb^® rabbit recombinant monoclonal antibody that detects I B phosphorylated on serine 32 and 36.

KLH-conjugated linear peptide corresponding to 10 amino acids surrounding phosphoserines 32 and 36 in human I B .

Quality Control Testing

Evaluated by Western Blotting with a construct containing N-terminal MBP-and C-terminal His-tagged recombinant I B protein fragment phosphorylated on serine 32 and 36.

Western Blotting Analysis (WB): A 1:10,000 dilution of this antibody detected a construct containing N-terminal MBP-and C-terminal His-tagged recombinant I B protein fragment phosphorylated on serine 32 and 36, but did not react with non-phosphorylated I B . (Phospho-I B construct: Courtesy of Dr. Jesse Rinehart, Yale University, School of Medicine).

Tested applications

Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected Phospho-I B -Ser32 and 36 in HeLa cells treated with TNF .

Affinity Binding Assay: A representative lot of this antibody bound Phospho-IkB -Ser32/36 peptide with a KD of 8.4 x 10-6 in an affinity binding assay.

Peptide Inhibition Assay: Target band detection in a construct containing construct containing N-terminal MBP-and C-terminal His-tagged recombinant I B protein fragment phosphorylated on serine 32 and 36 was prevented by preblocking of a representative lot with the immunogen phosphopeptide, but not the corresponding non-phosphopeptide.

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user

Evaluated by Western Blotting with a construct containing N-terminal MBP-and C-terminal His-tagged recombinant I B protein fragment phosphorylated on serine 32 and 36.

Western Blotting Analysis (WB): A 1:10,000 dilution of this antibody detected a construct containing recombinant I B protein fragment with MBP and His tags phosphorylated on serine 32 and 36, but did not react with non-phosphorylated I B. (Phospho-I B construct: Courtesy of Dr. Jesse Rinehart, Yale University, School of Medicine).

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. I B is known to inhibit the activity of dimeric NF- B/REL complexes by trapping Rel dimers in the cytoplasm through masking of their nuclear localization signals. In most resting cells, NF- B is sequestered in the cytoplasm in an inactive form associated with I B , which blocks the ability of NF- B to translocate to the nucleus. However, following exposure to pro-inflammatory cytokines, UV light, reactive oxygen species, or bacterial and viral toxins, I B is phosphorylated by I B kinases (IKK) on serine 32 and 36 that allows its polyubiquitination and proteasomal degradation. This unmasks the nuclear localization signal of NF- B and allows it to translocate to the nucleus where it binds to NF- B response elements in target genes and regulates their transcription. In the nucleus, it can bind to NF- B and facilitate its nuclear export. I B 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.

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.

300 μg/mL after reconstitution at 25 μL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

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.

ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany

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.