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CDKN1B 抗体

The 小鼠单克隆 anti-CDKN1B antibody is suitable to detect CDKN1B in samples from 人, 小鼠和大鼠. It has been validated for WB, IF, FACS 和 IHC (p). The antibody has been independently validated.

Immunocytochemistry validation image for anti-Cyclin-Dependent Kinase Inhibitor 1B (p27, Kip1) (CDKN1B) antibody (ABIN3025539) (CDKN1B 抗体)

ICC

Formalin-fixed, paraffin-embedded human colon carcinoma stained with p27 antibody (DCS-72. (CDKN1B 抗体)

IHC

3 images

(2 validations)

产品编号 ABIN3025539

发货至: 中国

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Quick Overview for CDKN1B 抗体 (ABIN3025539)

抗原

See all CDKN1B 抗体

CDKN1B (Cyclin-Dependent Kinase Inhibitor 1B (p27, Kip1) (CDKN1B))

适用

人, 小鼠, 大鼠

宿主

小鼠

克隆类型

单克隆

标记

This CDKN1B antibody is un-conjugated

应用范围

Western Blotting (WB), Immunofluorescence (IF), Flow Cytometry (FACS), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))

克隆位点

DCS-72-F6-KIP1-769

Discover our top product CDKN1B Primary Antibody

原理

p27 Antibody Cocktail

纯化方法

Protein G affinity chromatography

免疫原

Mouse recombinant protein (DCS-72.F6) and recombinant human protein (KIP1/769) were used as the immunogen for the p27 antibody cocktail.

亚型

IgG1, kappa
anti-Cyclin-Dependent Kinase Inhibitor 1B (p27, Kip1) (CDKN1B) (AA 150-198) antibody
CDKN1B 适用: 人, 小鼠, 大鼠 WB, ELISA, FACS, IHC (p), IF (cc), IF (p), IHC (fro) 宿主: 兔 Polyclonal unconjugated

anti-Cyclin-Dependent Kinase Inhibitor 1B (p27, Kip1) (CDKN1B) (AA 1-198) antibody
CDKN1B 适用: 人 WB, ELISA, IF, IHC (p), PLA 宿主: 小鼠 Monoclonal 4B4-E6 unconjugated

anti-Cyclin-Dependent Kinase Inhibitor 1B (p27, Kip1) (CDKN1B) antibody
KO Validated CDKN1B 适用: 人 WB, IHC, IF, IHC (p), IP, ICC, IHC (fro) 宿主: 兔 Polyclonal unconjugated

anti-Cyclin-Dependent Kinase Inhibitor 1B (p27, Kip1) (CDKN1B) antibody
CDKN1B 适用: 人, 小鼠, 大鼠, 猴 WB, IHC, IF, FACS 宿主: 小鼠 Monoclonal SX53G8 unconjugated

anti-Cyclin-Dependent Kinase Inhibitor 1B (p27, Kip1) (CDKN1B) antibody
KD Validated CDKN1B 适用: 人 WB, FACS, ICC 宿主: 兔 Monoclonal 23GB1825 unconjugated Recombinant Antibody

anti-Cyclin-Dependent Kinase Inhibitor 1B (p27, Kip1) (CDKN1B) (AA 1-198) antibody
CDKN1B 适用: 人 WB, IF 宿主: 小鼠 Polyclonal unconjugated

anti-Cyclin-Dependent Kinase Inhibitor 1B (p27, Kip1) (CDKN1B) (pThr187) antibody
CDKN1B 适用: 人, 小鼠, 大鼠 WB, IHC, ELISA, IF, ICC 宿主: 兔 Polyclonal unconjugated

anti-Cyclin-Dependent Kinase Inhibitor 1B (p27, Kip1) (CDKN1B) antibody
CDKN1B 适用: 人, 小鼠, 大鼠, 猴 WB, IHC, IF, FACS 宿主: 小鼠 Monoclonal DCS-72-F6 unconjugated

anti-Cyclin-Dependent Kinase Inhibitor 1B (p27, Kip1) (CDKN1B) antibody
CDKN1B 适用: 人 WB, IHC, ELISA, IF 宿主: 兔 Monoclonal 1G2 unconjugated Recombinant Antibody

anti-Cyclin-Dependent Kinase Inhibitor 1B (p27, Kip1) (CDKN1B) (AA 1-198) antibody
CDKN1B 适用: 人 WB, IF 宿主: 兔 Polyclonal unconjugated

应用备注

Optimal dilution of the p27 antibody to be determined by the researcher.
1. Staining of formalin-fixed tissues requires boiling tissue sections in 10 mM Citrate buffer, pH 6.0, for 10-20 min followed by cooling at RT for 20 min
2. The prediluted format is supplied in a dropper bottle and is optimized for use in IHC. After epitope retrieval step (if required), drip mAb solution onto the tissue section and incubate at RT for 30 min.

限制

仅限研究用
生效 #101901 (Western Blotting)

by

Johann-Friedrich-Blumenbach-Institute for Zoology and Anthropology, Department of Developmental Biology, Georg-August-University Göttingen

No.

#101901

日期

2017.12.04

抗原

P27

Lot Number

V2438-171009

Method validated

Western Blotting

Positive Control

NIH/3T3 mouse embryonic fibroblast cells overexpressing mVenus-tagged p27K, starved for 48h

Negative Control

Untransfected NIH/3T3 cells, starved for 48h

Notes

Passed. ABIN3025539 detects the ectopically expressed fusion protein as well as the endogenous protein by immunoblotting. Unspecific cross-reactivity is low.

Validation Images

Different volumes of total cellular lysates of either mVenus-p27K- transfected NIH/3T3 cells (1, 2, 3) or untransfected NIH/3T3 cells (4, 5) were loaded and the blot membrane was incubated with ABIN3025539 followed by incubation with anti-mouse IgG-linked to HRP and chemiluminescence detection. Expected molecular weight of p27 fused to mVenus is approximately 55kDa.
Full Methods
Primary Antibody

ABIN3025539

Secondary Antibody

anti-mouse IgG (whole molecule), HRP-linked (Sigma-Aldrich, A9044, lot 034M4761)

Full Protocol

Grow NIH/3T3 cells (ATCC, CRL-1658) in DMEM+GlutaMAX (Gibco, 31966-021, Lot 1852045) supplemented with fetal bovine serum (Gibco 270-106) and Pen/Strep (Gibco 15140), at 37°C and 5% CO₂ to 70% confluency.
Transfect cells with a plasmid encoding mVenus-tagged p27K^- (kindly provided by Toshio Kitamura, University of Tokyo; Oki et al., 2014) using EndofectinMax (GeneCopoeia) following the manufacturer's instructions.
Serum starve cells for 48h. Use untransfected NIH/3T3 cells starved for 48h as control.
Lyse cells in RIPA buffer (10mM PBS pH7.2, 2mM EDTA, 1% NP-40, 1% Triton X-100, protease inhibitors) at 4°C.
Denature total cellular lysates proteins in 1x SDS-sample buffer and separate proteins on a freshly cast denaturing 10% SDS-PAGE (Laemmli, 1970).
Transfer proteins onto 0.2µm Protran membrane (GE Healthcare, 10600004, A10043108) with a Western blotting system for 1h at 400A (Towbin et al., 1979).
Block the membrane in TBST (50mM Tris-HCl, pH7.4, 150mM NaCl, 0.2% Tween 20) containing 5% milk (blocking solution) for 60min at RT.
Incubate membrane with primary mouse anti-P27 antibody (antibodies-online, ABIN3025539, lot V2438-171009) diluted 1:250 in blocking solution ON at 4°C.
Incubate membrane with secondary anti-mouse IgG (whole molecule), HRP-linked (Sigma-Aldrich, A9044, lot 034M4761) diluted 1:5000 in TBST for 45min at RT.
Wash membrane with TBST for 30-45min at RT.
Reveal protein bands using Clarity Max Western ECL substrate (Bio-Rad, 1705062); image capture via Chemidoc Imaging System (BioRad).

Experimental Notes

ABIN3025539 reveals a protein band at the expected molecular weight of the mVenus-p27 fusion protein. The antibody does also detect a protein at the expected molecular weight of the endogenous, untagged p27.
生效 #102346 (Immunocytochemistry)

by

Johann-Friedrich-Blumenbach-Institute for Zoology and Anthropology, Department of Developmental Biology, Georg-August-University Göttingen

No.

#102346

日期

2017.12.04

抗原

P27

Lot Number

V2438-171009

Method validated

Immunocytochemistry

Positive Control

NIH/3T3 mouse embryonic fibroblast cells overexpressing mVenus-tagged p27K, starved for 48h

Negative Control

Untransfected NIH/3T3 cells, starved for 48h

Notes

Passed. ABIN3025539 detects the ectopically expressed fusion protein by immunocytochemistry. Unspecific cross-reactivity is low.

Validation Images

NIH/3T3 cells were transfected with a plasmid encoding mVenus fused to p27K- and the expressed fusion protein detected by Venus autofluorescence (B, green) and by immunocytology using ABIN3025539 and anti-mouse AF555 (A, red). C shows red and green channels merged with DAPI counterstain (blue).
Full Methods
Primary Antibody

ABIN3025539

Secondary Antibody

goat anti-mouse IgG (H+L) Alexa Fluor 555 (Invitrogen, A21422, lot 948498)

Full Protocol

Grow NIH/3T3 cells (ATCC, CRL-1658) in on cover slips in DMEM, 10% fetal bovine serum (Gibco 270-106), 5% penicillin/streptomycin (Gibco) at 37°C in 5% CO₂.
Transfect cells with a plasmid encoding mVenus-tagged p27K^- (kindly provided by Toshio Kitamura, University of Tokyo; Oki et al., 2014) using EndofectinMax (GeneCopoeia) following the manufacturer's instructions.
Serum starve cells for 48h. Use untransfected NIH/3T3 cells starved for 48h as control.
Fix cells in 3.7% paraformaldehyde (in PBS) for 15min at 4°C followed by incubation in 0.3% Triton X-100 for 10min.
Block unspecific binding sites in PBT (phosphate buffered saline (PBS) containing 1% bovine serum albumin, 0.5% Tween-20) for 1h at RT.
Incubate cells with primary mouse anti-P27 antibody (antibodies-online, ABIN3025539, lot V2438-171009) diluted 1:100 in PBS ON at 4°C.
Wash cells with TBST (50mM Tris-HCl, pH7.4, 150mM NaCl, 0.1% Tween 20) for 15min.
Incubate cells with secondary antibody goat anti-mouse IgG (H+L) Alexa Fluor 555 (Invitrogen, A21422, lot 948498) diluted 1:1000 in PBS and DAPI (4’,6-Diamidino-2-phenylindole; Sigma D-9542).
Image acquisition on Zeiss LSM 510 confocal microscope and processing using Adobe Photoshop 5.0.

Experimental Notes
状态

Liquid

浓度

0.2 mg/mL

缓冲液

0.2 mg/mL in 1X PBS with 0.1 mg/mL BSA (US sourced) and 0.05 % sodium azide

储存液

Sodium azide

注意事项

This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

储存条件

4 °C,-20 °C

储存方法

Store the p27 antibody cocktail at 2-8oC (with azide) or aliquot and store at -20oC or colder (without azide).
抗原

CDKN1B (Cyclin-Dependent Kinase Inhibitor 1B (p27, Kip1) (CDKN1B))

别名

p27

背景

Recognizes a 27 kDa protein, identified as the p27Kip1, a cell cycle regulatory mitotic inhibitor. It is highly specific and shows no cross-reaction with other related mitotic inhibitors. p27Kip1 functions as a negative regulator of G1 progression and has been proposed to function as a possible mediator of TGF- induced G1 arrest. p27Kip1 is a candidate tumor suppressor gene. This mAb co-precipitates cdk4 in complex p27Kip1 and is excellent for staining of formalin-fixed tissues.

UniProt

P46527

途径

Cell Division Cycle, Fc-epsilon Receptor Signaling Pathway, EGFR Signaling Pathway, Neurotrophin Signaling Pathway, Positive Regulation of Peptide Hormone Secretion, Negative Regulation of Hormone Secretion, Sensory Perception of Sound, Mitotic G1-G1/S Phases, DNA Replication, Positive Regulation of Endopeptidase Activity, Synthesis of DNA, Autophagy