Anti-p27 Kip1 Antibody, clone 1B10 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1429 - recombinant, expressed in HEK 293 cells
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human cerebellum (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. ZRB1429, Anti-p27 Kip1, clone 1B10 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Staining appears to be nuclear in subsets of cells in the granular layer of human cerebellum tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human cerebellum (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. ZRB1429, Anti-p27 Kip1, clone 1B10 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Staining appears to be nuclear in subsets of cells in the granular layer of human cerebellum tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human cerebellum (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. ZRB1429, Anti-p27 Kip1, clone 1B10 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Staining appears to be nuclear in subsets of cells in the granular layer of human cerebellum tissue sections.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cell line were performed using a 1:100 dilution of Cat. No. ZRB1429, Anti-p27 Kip1, clone 1B10 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 HeLa cell line were performed using a 1:100 dilution of Cat. No. ZRB1429, Anti-p27 Kip1, clone 1B10 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 HeLa cell line were performed using a 1:100 dilution of Cat. No. ZRB1429, Anti-p27 Kip1, clone 1B10 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 HUVEC were performed using a 1:100 dilution of Cat. No. ZRB1429, Anti-p27 Kip1, clone 1B10 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 HUVEC were performed using a 1:100 dilution of Cat. No. ZRB1429, Anti-p27 Kip1, clone 1B10 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 HUVEC were performed using a 1:100 dilution of Cat. No. ZRB1429, Anti-p27 Kip1, clone 1B10 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 Jurkat (Lane 1) and HeLa (Lane 2) cells were probed with Cat. No. ZRB1429, Anti-p27 Kip1, clone 1B10 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 p27 Kip1 (~30 kDa).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 100 mg/mL of non-specific control peptide (Run 1) or p27 Kip1 peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1429, Anti-p27 Kip1, clone 1B10 ZooMAb® Rabbit Monoclonal. The binding of this antibody to p27 Kip1 peptide displayed a KD of 2.5 x 10-8, whereas binding to non-specific control peptide displayed a KD of 9.2 x 10-4.
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 | 1B10, 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 | Designing Safer Chemicals Design for Energy Efficiency Design for Energy Efficiency Learn more about the Principles of Green Chemistry . |
| isotype | IgG |
| mol wt | calculated mol wt 22.07 kDa |
| observed mol wt ~30 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Protein ID accession no. | NP_004055  |
| 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) | monkey, mouse |
| 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. | P46527 |
| Application: | Quality Control Testing Evaluated by Western Blotting in Jurkat cell lysate. Western Blotting Analysis: A 1:1,000 dilution of this antibody detected p27 Kip1 in Jurkat cell lysate. Tested applications Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected p27 Kip1 in HeLa cell lysate. Affinity Binding Assay: A representative lot of this antibody bound in p27 Kip1 with a KD of 2.5 x 10-8 in an affinity binding assay. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected p27 Kip1 in HeLa cells and HUVEC. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected p27 Kip1 in human cerebellum tissue sections. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| Application: | Anti-p27 Kip1, clone 1B10 ZooMAb®, Cat. No. ZRB1429, is a recombinant Rabbit monoclonal antibody that detects p27 Kip1 and is tested for use in in Affinity Binding Assay, Immunocytochemistry, 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 18 amino acids from the N-terminal region of human Cyclin-dependent kinase inhibitor p27 (p27 Kip1). |
| 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 1B10 is a ZooMAb® Rabbit recombinant monoclonal antibody that specifically detects p27 Kip1. It targets an epitope within 18 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: | Cyclin-dependent kinase inhibitor 1B (UniProt: P46527; also known as Cyclin-dependent kinase inhibitor p27, p27Kip1) is encoded by the CDKN1B (also known as KIP1) gene (Gene ID: 1027) in human. p27Kip1 is a member of the universal cyclin-dependent kinase inhibitor family that serves as an important regulator of cell cycle progression. It inhibits the kinase activity of CDK2 bound to cyclin A. Its expression is regulated by cell contact inhibition and by specific growth factors. p27Kip1 is shown to be essential for the establishment of a G1 check point arrest after DNA damage. Phosphorylation of p27Kip1 by ATM kinase leads to its stabilization following DNA double strand breaks. In addition to its role as a cyclin-dependent kinase inhibitor p27Kip1 also acts as a tumor suppressor, regulator of drug resistance in solid tumors, and promoter of apoptosis. In quiescent cells it localizes in nucleus and cytoplasm and its levels are high that decline rapidly after stimulation with mitogens. Phosphorylation on serine 10 is the major site of phosphorylation in resting cells that takes place at the G0-G1 phase and leads to protein stability and facilitates nuclear export. Akt or Rsk- mediated phosphorylation on threonine 197 increases its interaction with 14-3-3 protein and allows it to translocates to the cytoplasm and promotes cell cycle progression. When phosphorylated on tyrosine 88 and 80, p27Kip1 translocates to the nucleus. Its nuclear localization signal is localized in amino acids 153-169. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Lloyd, RV et al., (1999). Am. J. Pathol. 154(2): 313-323; Cassimere, EK et al., (2016). PLoS One. 11(9); e0162806). |
| Storage Temp. | 2-8°C |