Anti-Skp2 Antibody, clone 1B13 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1784 - recombinant, expressed in HEK 293 cells
Synonym: S-phase kinase-associated protein 2;Cyclin-
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human Lung cancer (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. ZRB1784, Anti-Skp2, clone 1B13 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous/nuclear staining was observed in Human Lung cancer tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human Lung cancer (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. ZRB1784, Anti-Skp2, clone 1B13 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous/nuclear staining was observed in Human Lung cancer tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human Lung cancer (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. ZRB1784, Anti-Skp2, clone 1B13 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous/nuclear staining was observed in Human Lung cancer tissue sections.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of NIH 3T3 cells was performed using a 1:100 dilution of Cat. No. ZRB1784, Anti-Skp2, clone 1B13 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 and cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of NIH 3T3 cells was performed using a 1:100 dilution of Cat. No. ZRB1784, Anti-Skp2, clone 1B13 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 and cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of NIH 3T3 cells was performed using a 1:100 dilution of Cat. No. ZRB1784, Anti-Skp2, clone 1B13 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 and cytoplasm.
Western Blotting Enhanced Validation - Recombinant Antibody Technology Lysates from U2OS (Lane 1), HeLa (Lane 2), Cos-7 (Lane 3), and PC3 (Lane 4) cells were probed with Cat. No. ZRB1784, Anti-Skp2, clone 1B13 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 Skp2 (~48 kDa).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 100 mg/mL of non-specific control peptide (Run 1) or Skp2 peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1784, Anti-Skp2, clone 1B13 ZooMAb® Rabbit Monoclonal. The binding of this antibody to Skp2 peptide displayed a KD of 3.0 x 10-8, whereas binding to non-specific control peptide displayed a KD of 2.1 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 | 1B13, recombinant monoclonal |
| conjugate | unconjugated |
| description | recombinant, expressed in HEK 293 cells |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| epitope sequence | N-terminal |
| 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 47.76 kDa |
| observed mol wt ~48 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Protein ID accession no. | NP_005974  |
| purified by | using Protein A |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | monkey, mouse, human |
| species reactivity (predicted by homology) | bovine, porcine |
| 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. | Q13309 |
| Application: | Quality Control Testing Evaluated by Western Blotting in HeLa cell lysate. Western Blotting Analysis: A 1:1,000 dilution of this antibody detected Skp2 in HeLa cell lysate. Tested applications Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected Skp2 in U2OS, Cos-7, and PC3 cell lysates. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected Skp2 in human lung cancer tissue sections. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected Skp2 in NIH 3T3 cells. Affinity Binding Assay:: A representative lot of this antibody bound Skp2 with a KD of 3.0 x 10-8 in an affinity binding assay. 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 19 amino acids from the N-terminal region of human S-phase kinase-associated protein 2 (Skp2). |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS with 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 1B13 is a ZooMAb® Rabbit recombinant monoclonal antibody that specifically detects S-phase kinase-associated protein 2 (Skp2). It targets an epitope within 19 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: | S-phase kinase-associated protein 2 (UniProt: Q13309; also known as Cyclin-A/CDK2-associated protein p45, F-box protein Skp2, F-box/LRR-repeat protein 1, p45skp2) is encoded by the SKP2 (also known as FBXL1) gene (Gene ID: 6502) in human. Skp2 is the substrate recognition component of the SCF (Skp1 Cullin1 F-box (SCF) E3 ligase complex that mediates the ubiquitination and subsequent degradation of target proteins involved in cell cycle progression, signal transduction, and transcription. It plays an important role in governing cell cycle progression and cell survival by promoting the destruction of numerous tumor suppressor proteins, including p27, p21, p57, p130, and FOXO1. Hence, it functions as a proto-oncogene. It is shown to be essential for p27 degradation and thereby limits cells in G1 phase, prior to entry into S-phase. It specifically recognizes the phosphorylated form of p27. It also promotes ubiquitination and destruction of cadherin 1 (CDH1) in a CK1-dependent manner and hence regulates cell migration. Skp2 can undergo acetylation at lysine 68 and 71 that increases its stability through impairment of anaphase promoting complex/cyclosome (APC/C)-mediated proteolysis and promotes its cytoplasmic retention. Overexpression of Skp2 has been reported in a variety of human cancers, including lymphomas, prostate cancer, colorectal cancer, melanoma, pancreatic cancers, and nasopharyngeal carcinoma. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Inuzuka, H., et al. (2012). Cell. 150(1):179-93; Wang, Z., et al. (2012). Front. Oncol. 1(57); 57; Cardozo, T., and Pagano, M. (2004). Nat. Rev. Mol. Cell Biol. 5(9); 739-751). |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |