Overview

  • Product name

    Mouse p27Kip1 ELISA Kit
    See all p27 KIP 1 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 8 3.7%
    Inter-assay
    Sample n Mean SD CV%
    Overall 3 6.5%
  • Sample type

    Cell culture extracts, Tissue Extracts
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    7.5 pg/ml
  • Range

    15.6 pg/ml - 1000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture extracts 96 94% - 101%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse, Rat
  • Product overview

    Abcam’s p27Kip1 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of p27Kip1 protein in mouse tissue and cell extracts.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Notes

    Cyclin-dependent kinase inhibitor 1B (p27Kip1) is an enzyme inhibitor that is part of the Cip/Kip family of cyclin dependent kinase inhibitor proteins. p27Kip1 is a regulator of cell cycle progression involved at the G1 stage by binding to and preventing the activation of cyclin E-CDK2 or cyclin D-CDK4 complexes.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Mouse p27Kip1 Capture Antibody 1 x 600µl
    10X Mouse p27Kip1 Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent 4BR 1 x 6ml
    Mouse p27Kip1 Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 12ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • Function

    Important regulator of cell cycle progression. Involved in G1 arrest. Potent inhibitor of cyclin E- and cyclin A-CDK2 complexes. Forms a complex with cyclin type D-CDK4 complexes and is involved in the assembly, stability, and modulation of CCND1-CDK4 complex activation. Acts either as an inhibitor or an activator of cyclin type D-CDK4 complexes depending on its phosphorylation state and/or stoichometry.
  • Tissue specificity

    Expressed in all tissues tested. Highest levels in skeletal muscle, lowest in liver and kidney.
  • Involvement in disease

    Defects in CDKN1B are the cause of multiple endocrine neoplasia type 4 (MEN4) [MIM:610755]. Multiple endocrine neoplasia (MEN) syndromes are inherited cancer syndromes of the thyroid. MEN4 is a MEN-like syndrome with a phenotypic overlap of both MEN1 and MEN2.
  • Sequence similarities

    Belongs to the CDI family.
  • Domain

    A peptide sequence containing only AA 28-79 retains substantial Kip1 cyclin A/CDK2 inhibitory activity.
  • Post-translational
    modifications

    Phosphorylated; phosphorylation occurs on serine, threonine and tyrosine residues. Phosphorylation on Ser-10 is the major site of phosphorylation in resting cells, takes place at the G(0)-G(1) phase and leads to protein stability. Phosphorylation on other sites is greatly enhanced by mitogens, growth factors, cMYC and in certain cancer cell lines. The phosphorylated form found in the cytoplasm is inactivate. Phosphorylation on Thr-198 is required for interaction with 14-3-3 proteins. Phosphorylation on Thr-187, by CDK2 leads to protein ubiquitination and proteasomal degradation. Tyrosine phosphorylation promotes this process. Phosphorylation by PKB/AKT1 can be suppressed by LY294002, an inhibitor of the catalytic subunit of PI3K. Phosphorylation on Tyr-88 and Tyr-89 has no effect on binding CDK2, but is required for binding CDK4. Dephosphorylated on tyrosine residues by G-CSF.
    Ubiquitinated; in the cytoplasm by the KPC complex (composed of RNF123/KPC1 and UBAC1/KPC2) and, in the nucleus, by SCF(SKP2). The latter requires prior phosphorylation on Thr-187. Ubiquitinated; by a TRIM21-containing SCF(SKP2)-like complex; leads to its degradation.
    Subject to degradation in the lysosome. Interaction with SNX6 promotes lysosomal degradation.
  • Cellular localization

    Nucleus. Cytoplasm. Endosome. Nuclear and cytoplasmic in quiescent cells. AKT-or RSK-mediated phosphorylation on Thr-198, binds 14-3-3, translocates to the cytoplasm and promotes cell cycle progression. Mitogen-activated UHMK1 phosphorylation on Ser-10 also results in translocation to the cytoplasm and cell cycle progression. Phosphorylation on Ser-10 facilitates nuclear export. Translocates to the nucleus on phosphorylation of Tyr-88 and Tyr-89. Colocalizes at the endosome with SNX6 and this leads to lysosomal degradation.
  • Information by UniProt
  • Alternative names

    • AA408329
    • AI843786
    • Cdki1b
    • CDKN 1B
    • CDKN 4
    • CDKN1B
    • CDKN4
    • CDN1B_HUMAN
    • Cyclin Dependent Kinase Inhibitor 1B
    • Cyclin dependent kinase inhibitor p27
    • Cyclin-dependent kinase inhibitor 1B
    • Cyclin-dependent kinase inhibitor 1B (p27, Kip1)
    • Cyclin-dependent kinase inhibitor p27
    • Cyclin-dependent kinase inhibitor p27 Kip1
    • KIP 1
    • KIP1
    • MEN1B
    • MEN4
    • OTTHUMP00000195098
    • OTTHUMP00000195099
    • p27
    • p27 Kip1
    • P27-like cyclin-dependent kinase inhibitor
    • p27Kip1
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab208982 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of p27Kip1 were measured in duplicate and interpolated from the p27Kip1 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean p27Kip1 concentration was determined to be 496 pg/mL in NIH3T3 extract and 634 pg/mL in mouse skeletal muscle extract.

  • The concentrations of p27Kip1 in rat brain extract were measured in duplicate, interpolated from the mouse p27Kip1 standard curves and corrected for sample dilution. Undiluted samples are rat brain extract 62.5 μg/mL. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean p27Kip1 concentration was determined to be 1,077 pg/mL in rat brain extract.

Protocols

References

ab208982 has not yet been referenced specifically in any publications.

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