WB
Western blot analysis of p27 KIP 1 on different lysates with Rabbit anti-p27 KIP 1 antibody at 1/2000 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. 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 at 1/2000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:50000 dilution was used for 1 hour at room temperature.ICC/IF
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 (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 1/1000 dilution. The nuclear counter stain is DAPI (blue).ICC/IF
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 (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 1/1000 dilution. The nuclear counter stain is DAPI (blue).IHC
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 (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.IHC
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 (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.IHC
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 (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.FC
Flow cytometric analysis of p27 KIP 1 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (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 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Product Name | p27 KIP 1 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide within Human p27 KIP 1 aa 149-198 / 198. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCWB |
| WB:1:2000 ICC/IF:1:100-1:500 IHC:1:100-1:500 FC:1:50-1:100 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Affinity purified |
| Gene Symbol | CDKN1B |
|---|---|
| Gene Synonyms | KIP1 MEN4 CDKN4 MEN1B P27KIP1 |
| Gene Full Name | cyclin dependent kinase inhibitor 1B |
| Gene Summary | This gene encodes a cyclin-dependent kinase inhibitor, which shares a limited similarity with CDK inhibitor CDKN1A/p21. 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. The degradation of this protein, which is triggered by its CDK dependent phosphorylation and subsequent ubiquitination by SCF complexes, is required for the cellular transition from quiescence to the proliferative state. Mutations in this gene are associated with multiple endocrine neoplasia type IV (MEN4). [provided by RefSeq, Apr 2014] |
| Alternative Names | 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 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 Cyclin-dependent kinase inhibitor 1B (p27 Kip1) antibody Cyclin-dependent kinase inhibitor 1B antibody Cyclin-dependent kinase inhibitor p27 antibody Cyclin-dependent kinase inhibitor p27 Kip1 antibody KIP 1 antibody KIP1 antibody MEN1B antibody MEN4 antibody OTTHUMP00000195098 antibody OTTHUMP00000195099 antibody p27 antibody p27 Kip1 antibody P27-like cyclin-dependent kinase inhibitor antibody p27Kip1 antibody |
| Molecular Weight(MW) | 22kDa(Observed band size:27kDa) |
| Cellular Localization | Nucleus, Cytoplasm, Endosome. |
WB
Western blot analysis of p27 KIP 1 on different lysates with Rabbit anti-p27 KIP 1 antibody at 1/2000 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. 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 at 1/2000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:50000 dilution was used for 1 hour at room temperature.
ICC/IF
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 (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 1/1000 dilution. The nuclear counter stain is DAPI (blue).
ICC/IF
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 (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 1/1000 dilution. The nuclear counter stain is DAPI (blue).
IHC
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 (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.
IHC
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 (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.
IHC
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 (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.
FC
Flow cytometric analysis of p27 KIP 1 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (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 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Application Notes | WB:1:2000 ICC/IF:1:100-1:500 IHC:1:100-1:500 FC:1:50-1:100 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term. |
| Storage Buffer | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
