ET1608-61

p27 KIP 1 Recombinant Rabbit Monoclonal Antibody [SU37-04]

cat.: ET1608-61

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, FC, IP
Clonality:	Monoclonal
Clone number:	SU37-04

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 22 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human p27 KIP 1 aa 149-198 / 198.
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.
Subcellular location:	Nucleus, Cytoplasm, Endosome.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P FC IP	1:2,000 1:100-1:500 1:100-1:500 1:100-1:500 1:50-1:100 Use at an assay dependent concentration.
Uniprot #:	SwissProt: P46527 Human \| P46414 Mouse Unigene: 29897 Rat
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 (p27, Kip1) Cyclin-dependent kinase inhibitor 1B 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

Images

	Fig1: 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.
	Fig2: 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).
	Fig3: 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).

	Fig4: 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) for 20 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.
	Fig5: 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) for 20 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.
	Fig6: 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) for 20 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.
	Fig7: 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) for 20 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.

	Fig8: 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) for 20 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.
	Fig9: 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) for 20 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.
	Fig10: 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) for 20 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.
	Fig11: 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).

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