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ZRB1665

Sigma-Aldrich

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

enhanced validation
greener alternative

recombinant, expressed in HEK 293 cells

Synonym(s):

NF-kappa-B inhibitor alpha;I-kappa-B-alpha;IkB-alpha;IkappaB alpha;Major histocompatibility complex enhancer-binding protein MAD3

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About This Item

UNSPSC Code:
12352203
NACRES:
NA.43

biological source

rabbit

Quality Level

recombinant

expressed in HEK 293 cells

conjugate

unconjugated

antibody form

purified antibody

antibody product type

primary antibodies

clone

1I11, 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 ~45 kDa

purified by

using Protein A

species reactivity

human

species reactivity (predicted by homology)

mouse, rat

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
immunocytochemistry: suitable
inhibition assay: suitable
western blot: suitable

isotype

IgG

epitope sequence

N-terminal

Protein ID accession no.

UniProt accession no.

greener alternative category

shipped in

ambient

storage temp.

2-8°C

target post-translational modification

phosphorylation (pSer32/pSer36)

Gene Information

human ... NFKBIA(4792)

General description

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.

Specificity

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

Immunogen

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

Application

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).

Target 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. 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.

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.

Reconstitution

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.

Storage and Stability

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.

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.

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Storage Class Code

11 - Combustible Solids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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