概述

  • 产品名称
  • 描述
    兔多克隆抗体to Lin28
  • 经测试应用
    适用于: IP, Sandwich ELISA, ICC/IF, WBmore details
  • 种属反应性
    与反应: Mouse, Human
    预测可用于: Rat, Horse, Pig, Chimpanzee
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human Lin28.

    (Peptide available as ab64578.)

  • 阳性对照
    • This antibody gave a positive signal in the following Whole Cell Lysates: HUES-7 MEL-1 F9 ES-E14TG2a Mouse EG

性能

应用

Our Abpromise guarantee covers the use of ab63740 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
IP Use a concentration of 5 µg/ml.
Sandwich ELISA Use a concentration of 0.1 µg/ml. Can be paired for Sandwich ELISA with Mouse monoclonal [6D1F9] to Lin28 (ab76369). For sandwich ELISA, use this antibody as Detection at 0.1 µg/ml with Mouse monoclonal [6D1F9] to Lin28 (ab76369) as Capture.
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 30 kDa (predicted molecular weight: 23 kDa).

靶标

  • 功能
    Acts as a 'translational enhancer', driving specific mRNAs to polysomes and thus increasing the efficiency of protein synthesis. Its association with the translational machinery and target mRNAs results in an increased number of initiation events per molecule of mRNA and, indirectly, in stabilizing the mRNAs. Binds IGF2 mRNA, MYOD1 mRNA, ARBP/36B4 ribosomal protein mRNA and its own mRNA. Essential for skeletal muscle differentiation program through the translational up-regulation of IGF2 expression (By similarity). Acts as a suppressor of microRNA (miRNA) biogenesis by specifically binding the precursor let-7 (pre-let-7), a miRNA precursor. Acts by binding pre-let-7 and recruiting ZCCHC11/TUT4 uridylyltransferase, leading to the terminal uridylation of pre-let-7. Uridylated pre-let-7 miRNAs fail to be processed by Dicer and undergo degradation. Degradation of pre-let-7 in embryonic stem (ES) cells contributes to the maintenance of ES cells. In contrast, LIN28A down-regulation in neural stem cells by miR-125, allows the processing of pre-let-7. Specifically recognizes the 5'-GGAG-3' motif in the terminal loop of pre-let-7. Also recognizes and binds non pre-let-7 pre-miRNAs that contain the 5'-GGAG-3' motif in the terminal loop, leading to their terminal uridylation and subsequent degradation.
  • 组织特异性
    Expressed in embryonic stem cells (ES cells), placenta and testis.
  • 序列相似性
    Belongs to the lin-28 family.
    Contains 2 CCHC-type zinc fingers.
    Contains 1 CSD (cold-shock) domain.
  • 发展阶段
    Expressed in fetal liver. Expression decreases during differentiation of ES cells or upon induction of neuronal differentiation by retinoic acid.
  • 结构域
    The CSD domain is required for function in muscle differentiation.
  • 细胞定位
    Cytoplasm. Nucleus > nucleolus. Nucleolar localization observed in 10-15% of the nuclei in differentiated myotubes (By similarity). Shuttles between the cytoplasm and the nucleus. Localizes to cytoplasmic processing bodies and stress granules.
  • Information by UniProt
  • 数据库链接
  • 别名
    • AL024421 antibody
    • CSDD1 antibody
    • CSDD2 antibody
    • FLJ12457 antibody
    • Lin 28 antibody
    • Lin 28 homolog (C. elegans) antibody
    • Lin 28 homolog A (C. elegans) antibody
    • Lin 28 homolog A antibody
    • Lin 28 homolog antibody
    • Lin-28A antibody
    • Lin28 antibody
    • Lin28, C. elegans, homolog of, A antibody
    • LIN28A antibody
    • LN28A_HUMAN antibody
    • Protein lin-28 homolog A antibody
    • Protein lin-28 homolog B antibody
    • RNA binding protein lin 28 antibody
    • Tex17 antibody
    • ZCCHC1 antibody
    • Zinc finger CCHC domain containing 1 antibody
    • Zinc finger CCHC domain containing protein 1 antibody
    • Zinc finger CCHC domain-containing protein 1 antibody
    see all

图片

  • ICC/IF image of ab63740 stained mouse embryonic stem cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab63740, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue).
  • All lanes : Anti-Lin28 antibody (ab63740) at 1 µg/ml

    Lane 1 : MEL-1 (Human embryonic stem cell, male cell line) Whole Cell Lysate (ab27198)
    Lane 2 : F9 (Mouse embryonic carcinoma cell line) Whole Cell Lysate (ab27193)
    Lane 3 : E14Tg2a (Mouse embryonic stem cell line) Whole Cell Lysate
    Lane 4 : Mouse EG (TMAS Embryonic Germ Cells) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 23 kDa
    Observed band size : 30 kDa (why is the actual band size different from the predicted?)
    The 30 kDa band observed is comparable to molecular weights seen with other commercially available antibodies to Lin28.
  • All lanes : Anti-Lin28 antibody (ab63740) at 1 µg/ml

    Lane 1 : HUES7 (Human embryonic stem cell line) Whole Cell Lysate
    Lane 2 : F9 (Mouse embryonic carcinoma cell line) Whole Cell Lysate
    Lane 3 : E14Tg2a (Mouse embryonic stem cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 23 kDa
    Observed band size : 30 kDa (why is the actual band size different from the predicted?)


    Exposure time : 90 seconds

    This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab63740 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

  • Standard Curve for Lin28 (Analyte: Lin28 protein (His tag) (ab89225)); dilution range 1 pg/ml to 1 ug/ml using Capture Antibody Mouse monoclonal [6D1F9] to Lin28 (ab76369) at 0.2 ug/ml and Detector Antibody Rabbit polyclonal to Lin28 (ab63740) at 0.1 ug/ml.
  • Lin28 was immunoprecipitated using 0.5mg E14Tg2a whole cell extract, 5µg of Rabbit polyclonal to Lin28 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, E14Tg2a whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab63740.
    Secondary: Clean blot (HRP conjugate) at 1/1000 dilution.
    Band: 30kDa: Lin28.

文献

This product has been referenced in:
  • Szerlong HJ  et al. Proteomic characterization of the nucleolar linker histone H1 interaction network. J Mol Biol 427:2056-71 (2015). Read more (PubMed: 25584861) »
  • Enriquez VA  et al. High LIN28A Expressing Ovarian Cancer Cells Secrete Exosomes That Induce Invasion and Migration in HEK293 Cells. Biomed Res Int 2015:701390 (2015). WB . Read more (PubMed: 26583126) »

See all 5 Publications for this product

客户评价及客户问答

Application
Immunocytochemistry/ Immunofluorescence
Sample
Horse Cell (testis tissue)
Specification
testis tissue
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde
Username

Mr. Heejun Jung

Verified customer

提交于 Aug 19 2015

Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Reduced Denaturing (10% SDS)
Sample
Rat Cell lysate - whole cell (heart muscle in normal culture condtion)
Specification
heart muscle in normal culture condtion
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

提交于 Nov 03 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Wako target retrieval solution
Sample
Marmoset (common) Tissue sections (adult testis)
Specification
adult testis
Permeabilization
No
Fixative
Formaldehyde
Username

煜欽 林

Verified customer

提交于 Dec 19 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (embryonic stem cell)
Loading amount
1e+006 cells
Specification
embryonic stem cell
Gel Running Conditions
Reduced Denaturing (10% gel)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C
Username

Abcam user community

Verified customer

提交于 Aug 14 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (embryonic stem cell)
Specification
embryonic stem cell
Fixative
methanol:acetone 1:1
Permeabilization
Yes - 0.1% triton X-100
Blocking step
(agent) for 30 minute(s) · Concentration: 1% · Temperature: 16°C
Username

Abcam user community

Verified customer

提交于 Aug 14 2012

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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