p27 KIP 1 Recombinant Rabbit Monoclonal Antibody [SU37-04] - BSA and Azide free
Catalog# HA750157
p27 KIP 1 Recombinant Rabbit Monoclonal Antibody [SU37-04] - BSA and Azide free
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WB
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IF-Cell
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IF-Tissue
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IHC-P
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FC
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IP
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
p27 KIP 1 Recombinant Rabbit Monoclonal Antibody [SU37-04] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human p27 KIP 1 aa 149-198 / 198.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P, FC, IP
Target Molecular Weight
Predicted band size: 22 kDa
Positive Control
MCF7 cell lysate, Jurkat cell lysate, HeLa cell lysate, Neuro-2a cell lysate, C6 cell lysate, PC-12 cell lysate, A431, MCF-7, human tonsil tissue, human colon carcinoma tissue, human breast carcinoma tissue, mouse lung tissue, mouse colon tissue, mouse cerebellum tissue, rat brain tissue, Hela.
Conjugation
unconjugated
Clone Number
SU37-04
Product Features
Form
Liquid
Concentration
1mg/ml
Storage Instructions
Store at 2-8℃. Avoid freeze.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
Target
Function
Cyclin-dependent kinase inhibitor 1B (p27Kip1) is an enzyme inhibitor that in humans is encoded by the CDKN1B gene. It encodes a protein which belongs to the Cip/Kip family of cyclin dependent kinase (Cdk) inhibitor proteins. The encoded protein binds to and prevents the activation of cyclin E-CDK2 or cyclin D-CDK4 complexes, and thus controls the cell cycle progression at G1. It is often referred to as a cell cycle inhibitor protein because its major function is to stop or slow down the cell division cycle.
Background References
1. Gao K et al. HDGF-related protein-2 (HRP-2) acts as an oncogene to promote cell growth in hepatocellular carcinoma. Biochem Biophys Res Commun 458:849-55 (2015).
2. Wu J et al. Silencing of Kv1.5 Gene Inhibits Proliferation and Induces Apoptosis of Osteosarcoma Cells. Int J Mol Sci 16:26914-26 (2015).
Sequence Similarity
Belongs to the CDI family.
Tissue Specificity
Expressed in all tissues tested. Highest levels in skeletal muscle, lowest in liver and kidney.
Post-translational Modification
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 CDK1 and 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 (By similarity).
Subcellular Location
Nucleus, Cytoplasm, Endosome.
Synonyms
AA408329 antibody
AI843786 antibody
Cdki1b antibody
CDKN 1B antibody
CDKN 4 antibody
CDKN1B antibody
CDKN4 antibody
CDN1B_HUMAN antibody
Cyclin Dependent Kinase Inhibitor 1B antibody
Cyclin dependent kinase inhibitor p27 antibody
ExpandImages
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This data was developed using ET1608-61, the same antibody clone in a different buffer formulation.
Western blot analysis of p27 KIP 1 on different lysates with Rabbit anti-p27 KIP 1 antibody (ET1608-61) at 1/2,000 dilution.
Lane 1: MCF7 cell lysate
Lane 2: Jurkat cell lysate
Lane 3: HeLa cell lysate
Lane 4: Neuro-2a cell lysate
Lane 5: C6 cell lysate
Lane 6: PC-12 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 22 kDa
Observed band size: 27 kDa
Exposure time: 3 minutes 10 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1608-61) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature. -
This data was developed using ET1608-61, the same antibody clone in a different buffer formulation.
ICC staining of p27 KIP 1 in A431 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-61, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue). -
This data was developed using ET1608-61, the same antibody clone in a different buffer formulation.
ICC staining of p27 KIP 1 in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-61, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue). -
This data was developed using ET1608-61, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-p27 KIP 1 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-61, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
This data was developed using ET1608-61, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-p27 KIP 1 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-61, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
This data was developed using ET1608-61, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-p27 KIP 1 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-61, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
This data was developed using ET1608-61, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse lung tissue using anti-p27 KIP 1 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-61, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
This data was developed using ET1608-61, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse colon tissue using anti-p27 KIP 1 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-61, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
This data was developed using ET1608-61, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue using anti-p27 KIP 1 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-61, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
This data was developed using ET1608-61, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-p27 KIP 1 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-61, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
This data was developed using ET1608-61, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of p27 KIP 1 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1608-61, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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