LIN28A Monoclonal Antibody (14E6-4E6)

货号：MA1-016

规格：100 µg

价格：4070

产品类型：流式抗体

品牌：Thermo Fisher

抗原：LIN28A

物种：人/小鼠

宿主：小鼠

抗体亚型：其它

克隆号：14E6-4E6

荧光染料：其它

类型:	单抗	同型对照：	IgG2a
免疫原性：	Full-length human recombinant protein expressed in bacteria	用法：	1-3 µL(ChIP);1:100(Flow);1:50-1:200(ICC);1:50-1:200(IF);5 µg(IP);1 :1000(WB)

Product Specific InformationMA1-016 has been successfully used in Western blot, immunofluorescence, ChIP, flow cytometry, IHC (P) and immunoprecipitation applications on human and mouse samples.Target InformationLIN28A/B are expressed in various cancers and ES cells. LIN28 is a cytoplasmic protein, that acts as a 'translational enhancer', driving specific mRNAs to polysomes and thus increasing the efficiency of protein synthesis. LIN28 is a negative regulator of microRNA (miRNA) biogenesis by specifically binding the precursor of let-7 (pre-let-7). Let-7 miRNAs have been implicated in repression of oncogenes such as ras and myc. LIN28 expression decreases during differentiation of ES cells or upon induction of neuronal differentiation by retinoic acid.For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

数据

LIN28A Antibody (MA1-016) in WBWestern blot analysis of LIN28 was performed by loading 75 µg of various whole cell lysates and 10 µl of PageRuler Prestained Protein Ladder (Product # 26616) onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with a LIN28 monoclonal antibody (Product # MA1-016) at a dilution of 1:1000 overnight at 4°C on a rocking platform, washed in TBS-0.1%Tween-20, and probed with a goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:20,000 for 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34075). LIN28A Antibody (MA1-016) in FlowFlow cytometric analysis of Lin28 (blue histogram) on HEL 11.4 induced IPS cells. To generate single cells suspensions, colonies were treated with TrypLE cell dissociation enzyme for 5 minutes at 37°C. Cells were incubated with a Lin28 monoclonal antibody (Product # MA1-016) or mouse IgG (green histogram) at a dilution of 1:100 for 1 hour on ice, washed with PBS + 5% fetal calf serum (FACS buffer), and incubated with a fluorescein-conjugated secondary antibody at a dilution of 1:200 for 30 minutes on ice. Cells were washed with cold FACS buffer, resuspended in 500 µL of FACS buffer containing 10 µL of 4% paraformaldehyde, and analyzed on a flow cytometer. LIN28A Antibody (MA1-016) in IFImmunofluorescent analysis of Lin28 (green) in HEL 11.4 induced IPS cells grown for a few days on Matrigel-coated chamber slides. Cells fixed in 4% paraformaldehyde were permeabilized with 0.1% Triton X-100 for 15 minutes at room temperature. Cells were probed with a Lin28 monoclonal antibody (Product # MA1-016) at a dilution of 1:200 overnight at 4°C, washed with PBST, and incubated with a fluorescein-conjugated secondary antibody at a dilution of 1:100 for 1 hour at room temperature. Nuclei (blue) were stained with DAPI and cells were analyzed by fluorescence microscopy at 20X magnification. LIN28A Antibody (MA1-016) in IHC (P)Immunohistochemistry analysis of LIN28 showing staining in the nucleus and cytoplasm of paraffin-embedded mouse testis tissue (right) compared with a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a LIN28 monoclonal antibody (Product # MA1-016) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

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参考文献：

1.Stem cell reports Conditionally Stabilized dCas9 Activator for Controlling Gene Expression in Human Cell Reprogramming and Differentiation. "MA1-016 was used in immunocytochemistry to demonstrate that dCas9 activator controls human pluripotent stem cell differentiation into endodermal lineages" AuthorsBalboa D,Weltner J,Eurola S,Trokovic R,Wartiovaara K,Otonkoski T

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