Anti-NFkB2 (p100/p52) Antibody, clone 5F3 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1250 - recombinant, expressed in HEK 293 cells
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human small intestine (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1250, Anti-NFkB2 (p100/p52), clone 5F3 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in glandular cells and lamina propria of human small intestine tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human small intestine (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1250, Anti-NFkB2 (p100/p52), clone 5F3 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in glandular cells and lamina propria of human small intestine tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human small intestine (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1250, Anti-NFkB2 (p100/p52), clone 5F3 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in glandular cells and lamina propria of human small intestine tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human testis intestine (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1250, Anti-NFkB2 (p100/p52), clone 5F3 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/perinuclear/membranous staining was observed in seminiferous tubules and Leydig cells of human testis tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human testis intestine (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1250, Anti-NFkB2 (p100/p52), clone 5F3 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/perinuclear/membranous staining was observed in seminiferous tubules and Leydig cells of human testis tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human testis intestine (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1250, Anti-NFkB2 (p100/p52), clone 5F3 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/perinuclear/membranous staining was observed in seminiferous tubules and Leydig cells of human testis tissue sections.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HepG2 cell line was performed using a 1:100 dilution of Cat. No. ZRB1250, Anti-NFkB2 (p100/p52), clone 5F3 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HepG2 cell line was performed using a 1:100 dilution of Cat. No. ZRB1250, Anti-NFkB2 (p100/p52), clone 5F3 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HepG2 cell line was performed using a 1:100 dilution of Cat. No. ZRB1250, Anti-NFkB2 (p100/p52), clone 5F3 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Western Blotting Enhanced Validation - Recombinant Antibody Technology Lysates from HeLa cells (Lane 1), Jurkat cells (Lane 2), and HepG2 cells (Lane 3) were probed with Cat. No. ZRB1250, Anti-NFkB2 (p100/p52), clone 5F3 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. Arrows indicate NFkB2 (p100/p52) (~110 and 52 kDa).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 100 mg/mL of non-specific control peptide (Run 1) or NFkB2 peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1250, Anti-NFkB2 (p100/p52), clone 5F3 ZooMAb® Rabbit Monoclonal. The binding of this antibody to NFkB2 peptide displayed a KD of 1.5 x 10-8, whereas binding to non-specific control peptide displayed a KD of 5.3 x 10-3.
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 | 5F3, monoclonal |
| recombinant monoclonal | |
| conjugate | unconjugated |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| 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 ~96.75 kDa |
| observed mol wt ~110 and 50 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human |
| species reactivity (predicted by homology) | bovine, monkey |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| immunocytochemistry: suitable | |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable | |
| western blot: suitable | |
| UniProt accession no. | Q00653  |
| Application: | Quality Control Testing Evaluated by Western Blotting in HeLa cell lysate. Western Blotting Analysis: A 1:1,000 dilution of this antibody detected NFkB2 (p100/p52) in HeLa cell lysate. Tested Applications Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected NFkB2 (p100/p52) in Jurkat and HepG2 cell lysates. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected NFkB2 (p100/p52) in human small intestine and human testis tissue sections. Affinity Binding Assay: A representative lot of this antibody bound NFkB2 peptide with a KD of 4.5 x 10-8 in an affinity binding assay. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected NFkB2 (p100/p52) in HepG2 cells. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| Application: | Anti-NFkB2 (p100/p52), clone 5F3 ZooMAb®, Cat. No. ZRB1250, is a recombinant Rabbit monoclonal antibody that targets p100/p52 of NFkB2 and is tested in Affinity Binding Assay, Immunocytochemistry, and Immunohistochemistry (Paraffin), and Western Blotting |
| 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 16 amino acids from the N-terminal region of human Nuclear factor NF-kappa-B p100 subunit. |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant mouse monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. 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 5F3 is a ZooMAb® rabbit recombinant monoclonal antibody that specifically detects NFkB2 (p100/p52). It targets an epitope within 16 amino acids from the N-terminal region. |
| 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: | Nuclear factor NF- kB p100 subunit (UniProt: Q00653; also known as DNA-binding factor KBF2, H2TF1, Lymphocyte translocation chromosome 10 protein, Nuclear factor of kappa light polypeptide gene (Gene ID: 4791) in human. NF kB is a pleiotropic transcription factor present in almost all cell types and is the endpoint of a series of signal transduction events that are initiated by a vast array of stimuli related to many biological processes such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. Five members of the NF- kB family have been identified that associate to form various homodimeric or heterodimeric combinations. They share a highly conserved Rel homology domain (RHD), which is responsible for DNA binding, dimerization, and interaction with IkBs. In most resting cells, NF- kB is sequestered in the cytoplasm in an inactive form associated with inhibitory molecules, which blocks the ability of NF- kB to translocate to the nucleus and bind to DNA. In a conventional activation pathway, IkB is phosphorylated and degraded thereby liberating NF- kB to translocate to the nucleus. In a non-canonical activation pathway, CHUK/IKKA homodimer phosphorylates NFkB2/p100 associated with RelB, inducing its proteolytic processing to NFkB2/p52 and the formation of RelB-p52 complex that serves as a transcriptional activator. However, the p52-p52 homodimer acts as a transcriptional repressor. NFkB2 has dual functions such as cytoplasmic retention of attached NF- kB proteins by p100 and generation of p52 by a co-translational processing. p52 is shown to bind to the kB consensus sequence 5′-GGRNNYYCC-3′, located in the enhancer region of genes involved in immune response and acute phase reactions. Clone 5F3 detects both the p100 and p52 units. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. |
| WGK Germany | WGK 1 |
| Storage Temp. | 2-8°C |