Anti-p16 Antibody, clone D25 ZooMAb® Mouse Monoclonal
SIGMA/ZMS1072 - recombinant, expressed in HEK 293 cells
Product Type: Chemical
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells was performed using a 1:100 dilution of Cat. No. ZMS1072, Anti-p16, clone D25 ZooMAb® Mouse Monoclonal and visualized with a Goat Anti-Mouse 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 HeLa cells was performed using a 1:100 dilution of Cat. No. ZMS1072, Anti-p16, clone D25 ZooMAb® Mouse Monoclonal and visualized with a Goat Anti-Mouse 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 HeLa cells was performed using a 1:100 dilution of Cat. No. ZMS1072, Anti-p16, clone D25 ZooMAb® Mouse Monoclonal and visualized with a Goat Anti-Mouse 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 Recombinant human p16 (CDKN2A) protein was probed with Cat. No. ZMS1072, Anti-p16, clone D25 ZooMAb® Mouse Monoclonal (1:10,000 dilution). Proteins were visualized using a Goat Anti-Mouse IgG1 secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates p16 (~16 kDa).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 10 mg/mL of non-specific GST protein (Run 1) or recombinant p16 protein (Run 2) were analyzed by affinity binding assay using Cat. No. ZMS1072, Anti-p16, clone D25 ZooMAb® Mouse Monoclonal. The binding of this antibody to p16 protein displayed a KD of 2.5 x 10-6 whereas binding to non-specific GST protein displayed a KD of 2.8 x 10-3.
Flow Cytometry Enhanced Validation - Recombinant Antibody Technology Staining of one million HeLa cells was performed using 1 μg of Cat. No. ZMS1072, Anti-p16, clone D25 ZooMAb® Mouse Monoclonal (Yellow histogram) or the equivalent amount of a Mouse IgG1 isotype control (Grey histogram), followed by a PE-conjugated Goat Anti-Mouse IgG1 secondary antibody.
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 | mouse |
| clone | D25, recombinant monoclonal |
| conjugate | unconjugated |
| description | recombinant, expressed in HEK 293 cells |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| epitope sequence | Unknown |
| 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 | IgG1 |
| mol wt | calculated mol wt 16.53 kDa |
| observed mol wt ~16 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 |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| flow cytometry: suitable | |
| immunocytochemistry: suitable | |
| western blot: suitable | |
| UniProt accession no. | P42771  |
| Application: | Quality Control Testing Evaluated by Western Blotting with human recombinant p16 (CDKN2A).. Western Blotting Analysis: A 1:10,000 dilution of this antibody detected human recombinant p16 (CDKN2A). Tested applications Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected p16 in HeLa cells. Flow Cytometry Analysis: 1 μg from a representative lot detected p16 in Hela cells. Affinity Binding Assay: A representative lot of this antibody bound recombinant human p16 (CDKN2A) with a KD of 2.5 x 10-6 in an affinity binding assay. Flow Cytometry Analysis: 1 μg from a representative lot detected p16 (CDKN2A) 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 |
| Application: | Anti-p16, clone D25 ZooMAb®, Cat. No. ZMS1072, is a recombinant Mouse monoclonal antibody that detects p16 (CDKN2A) and is tested for use in Affinity Binding Assay, Flow Cytometry, Immunocytochemistry, 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. Learn more about ZooMAb here. |
| Immunogen: | Recombinant human p16 fusion protein. |
| 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 D25 is a ZooMAb® mouse recombinant monoclonal antibody that specifically detects Cyclin-dependent kinase inhibitor 2A (p16). |
| 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: | Cyclin-dependent kinase inhibitor 2A (UniProt: P42771; also known as Cyclin-dependent kinase 4 inhibitor A, CDK4I, Multiple tumor suppressor 1, MTS-1, p16-INK4a, p16-INK4, p16INK4A) is encoded by the CDKN2A (also known as CDKN2, MTS1) gene (Gene ID: 1029) in human. p16 serves as a negative regulator of cell proliferation by strongly interacting with CDK4 and CDK6 during the G1-to S-phase transition and blocking the phosphorylation of retinoblastoma (Rb) protein. Hence, it is considered as a tumor suppressor protein. Hypo-phosphorylated Rb protein forms complexes with E2F family of transcription factors, which sequesters E2Fs and prevent their access to the promoters of proliferation-associated genes. During cell cycle, in the late G1 phase CDK4 and CDK6 hyper-phosphorylate Rb protein that commits cells to enter into S phase. Binding of p16 maintains Rb in a hypo-phosphorylated state. CDKN2A gene is reported to be one of the most frequently mutated genes and is reported to be inactivated in about 50% of all human cancers. This ZooMAb recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Li, J., et al. (2011). Biochemistry. 50(25); 5566-5582; Sherr, CJ., and Roberts, JM (2004). Genes Dev. 18(22); 2699-2711; Ortega, S., et al. (2002). Biochim. Biophys. Acta. 1602(1); 73-87). |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |