Anti-p-IκBα-Ser32 Antibody, clone 11D19 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1554 - recombinant, expressed in HEK 293 cells
Product Type: Chemical
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells was performed using a 1:1,000 dilution of Cat. No. ZRB1554, Anti-p-IκBα-Ser32, clone 11D19 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa FluorZooMAb® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the Nucleus and cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells was performed using a 1:1,000 dilution of Cat. No. ZRB1554, Anti-p-IκBα-Ser32, clone 11D19 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa FluorZooMAb® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the Nucleus and cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells was performed using a 1:1,000 dilution of Cat. No. ZRB1554, Anti-p-IκBα-Ser32, clone 11D19 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa FluorZooMAb® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the Nucleus and cytoplasm.
Western Blotting Enhanced Validation - Recombinant Antibody Technology Lysates from untreated HeLa cells (Lane 1) and HeLa cells treated with TNF-α (100 ng/mL)(Lane 2) were probed with Cat. No. ZRB1554, Anti-p-IκBα-Ser32, clone 11D19 ZooMAb® Rabbit Monoclonal (1:1,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates IkBα (~40 kDa).
Western Blotting Enhanced Validation - Recombinant Antibody Technology Constructs containing GST-tagged non-phosphorylated recombinant IkBa protein fragment (Lane 1) and GST-tagged recombinant IkBa protein fragment phosphorylated on Ser32 (Lane 2) were probed with Cat. No. ZRB1554, Anti-p-IκBα-Ser32, clone 11D19 ZooMAb® Rabbit Monoclonal ( (1:1,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates IkBα (~40 kDa). (Constructs: Courtesy of Dr. Jesse Rinehart, Yale University, School of Medicine).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 100 mg/mL of non-specific control peptide (Run1) or phosphor-IkBa-Ser32 peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1554, Anti-p-IκBα-Ser32, clone 11D19 ZooMAb® Rabbit Monoclonal. The binding of this antibody to phosphor-IkBa-Ser32 peptide displayed a KD of 6.9 x 10-7 whereas binding to non-specific control peptide displayed a KD of 1.5 x 10-3.
Flow Cytometry Enhanced Validation - Recombinant Antibody Technology Staining of one million Hela cells was performed using 0.1 μg of Cat. No. ZRB1554, Anti-p-I B -Ser32, clone 11D19 ZooMAb® Rabbit Monoclonal (Yellow histogram) or the equivalent amount of a Rabbit IgG isotype control (Grey histogram), followed by a PE-conjugated Donkey Anti-Rabbit IgG secondary antibody.
Peptide Inhibition Assay Enhanced Validation - Recombinant Antibody Technology Constructs containing GST-tagged recombinant IkBa protein fragment phosphorylated on Ser32 were probed with Cat. No. ZRB1554, Anti-p-IκBα-Ser32, clone 11D19 ZooMAb® Rabbit Monoclonal ( (1:1,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates IkBα (~40 kDa). (Construct: Courtesy of Dr. Jesse Rinehart, Yale University, School of Medicine). Lane 1: Incubated without any peptide. Lane 2: Incubated with phosphorylated IkBa-Ser32 peptide Lane 3: Incubated with a non-phosphorylated IkBa-Ser32 peptide.
12 Principles of Green Chemistry: Principle 1—Waste Prevention. This product has a smaller environmental footprint through reduction of waste in the manufacturing process.
12 Principles of Green Chemistry: Principle 4—Designing Safer Chemicals
Chemical products should be designed to affect their desired function while minimizing their toxicity.
Chemical products should be designed to affect their desired function while minimizing their toxicity.
12 Principles of Green Chemistry: Principle 6—Design for Energy Efficiency. This product was re-engineered to utilize less energy in the manufacturing process.
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| biological source | rabbit |
| clone | 11D19, recombinant monoclonal |
| conjugate | unconjugated |
| description | recombinant, expressed in HEK 293 cells |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| epitope sequence | N-terminus |
| form | lyophilized |
| greener alternative category | Aligned |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry . |
| isotype | IgG |
| mol wt | calculated mol wt 35.61 kDa |
| observed mol wt ~40 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Protein ID accession no. | NP_065390  |
| purified by | using Protein A |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human |
| species reactivity (predicted by homology) | porcine, monkey, feline, canine, bovine |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| flow cytometry: suitable | |
| immunocytochemistry: suitable | |
| inhibition assay: suitable | |
| western blot: suitable | |
| UniProt accession no. | P25963 |
| 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. (Phospho- IkBa construct: Courtesy of Dr. Jesse Rinehart, Yale University, School of Medicine). 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 |
| 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. |
| 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. |
| General description: | 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. |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| 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. |
| Specificity: | Clone 11D19 is a ZooMAb® rabbit recombinant monoclonal antibody that specifically detects IkBa phosphorylated on serine 32. |
| 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. |
| 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. 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. |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |