Recombinant Anti-p27 KIP 1 antibody [EPFHCR16] (ab92741)
![Western blot - Anti-p27 KIP 1 antibody [EPFHCR16] (ab92741)](https://www.abcam.com/ps/products/92/ab92741/Images/ab92741-477278-anti-p27-kip-1-antibody-epfhcr16-western-blot-wildtype-raw-2647-cdkn1b-knockout-raw-2647.jpg "Western blot - Anti-p27 KIP 1 antibody [EPFHCR16] (ab92741)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPFHCR16] to p27 KIP 1
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat
Related conjugates and formulations
Overview
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Product name
Anti-p27 KIP 1 antibody [EPFHCR16]
See all p27 KIP 1 primary antibodies -
Description
Rabbit monoclonal [EPFHCR16] to p27 KIP 1 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-Pmore details
Unsuitable for: Flow Cyt,ICC/IF or IP -
Species reactivity
Reacts with: Mouse, Rat -
Immunogen
Recombinant fragment corresponding to Mouse p27 KIP 1.
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Positive control
- WB: NIH3T3, C6 and Neuro 2a cell lysates. IHC-P: Rat testis, mouse testis, lung and thymus tissues.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Human: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 0.31% Sodium citrate, 0.175% Sodium chloride, 0.0172% EDTA, 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPFHCR16 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab92741 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 |
|---|---|---|
| WB |
1/1000 - 1/5000. Detects a band of approximately 27 kDa (predicted molecular weight: 22 kDa).
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| IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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| Notes |
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WB
1/1000 - 1/5000. Detects a band of approximately 27 kDa (predicted molecular weight: 22 kDa). |
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IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Target
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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
modificationsPhosphorylated; 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
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Database links
- Entrez Gene: 12576 Mouse
- Entrez Gene: 83571 Rat
- SwissProt: P46414 Mouse
- Unigene: 2958 Mouse
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Alternative names
- AA408329 antibody
- AI843786 antibody
- Cdki1b antibody
see all
Images
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Western blot - Anti-p27 KIP 1 antibody [EPFHCR16] (ab92741)All lanes : Anti-p27 KIP 1 antibody [EPFHCR16] (ab92741) at 1/1000 dilution
Lane 1 : Wild-type RAW 264.7 cell lysate
Lane 2 : CDKN1B knockout RAW 264.7 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 22 kDa
Observed band size: 28 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-p27 KIP 1 antibody [EPFHCR16] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab92741 was shown to bind specifically to p27 KIP 1. A band was observed at 28 kDa in wild-type RAW 264.7 cell lysates with no signal observed at this size in CDKN1B knockout cell line ab281619 (knockout cell lysate ab282970). To generate this image, wild-type and CDKN1B knockout RAW 264.7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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![Western blot - Anti-p27 KIP 1 antibody [EPFHCR16] (ab92741)](https://www.abcam.com/ps/products/92/ab92741/Images/ab92741-107959-anti-p27-kip-1-antibody-epfhcr16-western-blot.jpg)
Western blot - Anti-p27 KIP 1 antibody [EPFHCR16] (ab92741)All lanes : Anti-p27 KIP 1 antibody [EPFHCR16] (ab92741) at 1/1000 dilution
Lane 1 : NIH/3T3 cell lysate
Lane 2 : C6 cell lysate
Lane 3 : Neuro-2a cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti-rabbit HRP-conjugated at 1/2000 dilution
Predicted band size: 22 kDa
Observed band size: 27 kDa why is the actual band size different from the predicted? -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p27 KIP 1 antibody [EPFHCR16] (ab92741)](https://www.abcam.com/ps/products/92/ab92741/Images/ab92741-293219-anti-p27-kip-1-antibody-epfhcr16-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p27 KIP 1 antibody [EPFHCR16] (ab92741)Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling p27 KIP 1 with ab92741 at 1/5000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining in Leydig and Sertoli cells and Leydig cells of mouse testis is observed (PMID: 10098522). Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody.
Heat mediated antigen retrieval was performed using Universal HIER antigen retrieval reagent (10X) (ab208572).
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p27 KIP 1 antibody [EPFHCR16] (ab92741)](https://www.abcam.com/ps/products/92/ab92741/Images/ab92741-293220-anti-p27-kip-1-antibody-epfhcr16-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p27 KIP 1 antibody [EPFHCR16] (ab92741)Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling p27 KIP 1 with ab92741 at 1/5000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining in Leydig and Sertoli cells and Leydig cells of rat testis is observed (PMID: 10098522). Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody.
Heat mediated antigen retrieval was performed using Universal HIER antigen retrieval reagent (10X) (ab208572).
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p27 KIP 1 antibody [EPFHCR16] (ab92741)](https://www.abcam.com/ps/products/92/ab92741/Images/ab92741-107958-anti-p27-kip-1-antibody-epfhcr16-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p27 KIP 1 antibody [EPFHCR16] (ab92741)ab92741 at 1/100 dilution staining p27 KIP in Mouse lung by Immunohistochemistry, Paraffin-embedded tissue.
Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p27 KIP 1 antibody [EPFHCR16] (ab92741)](https://www.abcam.com/ps/products/92/ab92741/Images/ab92741-107957-anti-p27-kip-1-antibody-epfhcr16-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p27 KIP 1 antibody [EPFHCR16] (ab92741)ab92741 at 1/100 dilution staining p27 KIP in Mouse thymus by Immunohistochemistry, Paraffin-embedded tissue.
Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.
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![Anti-p27 KIP 1 antibody [EPFHCR16] (ab92741)](https://www.abcam.com/ps/products/92/ab92741/Images/ab92741-6-benefits-of-recombinant-antibodies.png)
Anti-p27 KIP 1 antibody [EPFHCR16] (ab92741)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (15)
ab92741 has been referenced in 15 publications.
- Xia RM et al. LHPP-Mediated Histidine Dephosphorylation Suppresses the Self-Renewal of Mouse Embryonic Stem Cells. Front Cell Dev Biol 9:638815 (2021). PubMed: 33796530
- Zhang W et al. IPO5 promotes the proliferation and tumourigenicity of colorectal cancer cells by mediating RASAL2 nuclear transportation. J Exp Clin Cancer Res 38:296 (2019). PubMed: 31288861
- Lu Z et al. Dracorhodin perchlorate induces apoptosis and G2/M cell cycle arrest in human esophageal squamous cell carcinoma through inhibition of the JAK2/STAT3 and AKT/FOXO3a pathways. Mol Med Rep 20:2091-2100 (2019). PubMed: 31322237
- Wang M et al. Nongenetic optical modulation of neural stem cell proliferation and neuronal/glial differentiation. Biomaterials 225:119539 (2019). PubMed: 31622821
- Fu D et al. Hsa_circ_0057481 promotes laryngeal cancer proliferation and migration by modulating the miR-200c/ZEB1 axis. Int J Clin Exp Pathol 12:4066-4076 (2019). PubMed: 31933802