重组Anti-Lin28A抗体[EPR24286-100] (ab279647)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR24286-100] to Lin28A
- Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IF, IHC-Fr
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-Lin28A抗体[EPR24286-100]
参阅全部 Lin28A 一抗 -
描述
兔单克隆抗体[EPR24286-100] to Lin28A -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IF, IHC-Frmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: NCCIT, JAR and F9 whole cell lysate. IHC-P: Mouse testis and ovary tissue. Rat testis tissue. IHC-Fr: Mouse testis tissue. ICC/IF: F9 and NCCIT cells. Flow Cyt (intra): F9 and NCCIT cells. IP: F9 and NCCIT whole cell lysate.
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常规说明
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
存储溶液
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR24286-100 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab279647于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
---|---|---|
Flow Cyt (Intra) |
1/500.
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WB |
1/1000. Predicted molecular weight: 23 kDa.
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IP |
1/30.
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|
IHC-P |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/500.
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|
IHC-Fr |
1/100.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20) |
说明 |
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Flow Cyt (Intra)
1/500. |
WB
1/1000. Predicted molecular weight: 23 kDa. |
IP
1/30. |
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/500. |
IHC-Fr
1/100. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20) |
靶标
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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
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数据库链接
- Entrez Gene: 79727 Human
- Entrez Gene: 83557 Mouse
- Entrez Gene: 500562 Rat
- Omim: 611043 Human
- SwissProt: Q9H9Z2 Human
- SwissProt: Q8K3Y3 Mouse
- Unigene: 86154 Human
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别名
- CSDD1 antibody
- FLJ12457 antibody
- LIN 28 antibody
see all
图片
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All lanes : Anti-Lin28A antibody [EPR24286-100] (ab279647) at 1/5000 dilution
Lane 1 : NCCIT (human pluripotent embryonic carcinoma epithelial cell) whole cell lysate
Lane 2 : JAR (human placenta choriocarcinoma epithelial cell) whole cell lysate
Lane 3 : HEK-293T (human embryonic kidney epithelial cell) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 23 kDa
Observed band size: 25 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
The observed MW is consistent with what has been described in the literature (PMID:25479749)
Negative control: HEK-293T (PMID: 25479749)
Exposure time: 10 seconds
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NCCIT (human pluripotent embryonic carcinoma epithelial cell line) cells labeling Lin28A with ab279647 at 1/500 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing nucleolar and cytoplasmic staining in NCCIT cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (Red). The nuclear counterstain was DAPI (Blue).
Negative control: HEK-293T (PMID: 25479749).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
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Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HEK-293T (human embryonic kidney epithelial cell, Left panel) / NCCIT (human pluripotent embryonic carcinoma epithelial cell, Right panel) cells labeling Lin28A with ab279647 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Negative control: HEK-293T (PMID: 25479749).
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Lin28A was immunoprecipitated from 0.35 mg NCCIT (human pluripotent embryonic carcinoma epithelial cell), whole cell lysate 10 µg with ab279647 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab279647 at 1/5000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/10000 dilution.
Lane 1: NCCIT whole cell lysate 10 µg
Lane 2: ab279647 IP in NCCIT whole cell lysate 10 µg
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab279647 in NCCIT whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 8 seconds
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Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling Lin28A with ab279647 at 1/1000 dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse undifferentiated spermatogonia (PMID: 32094118). The section was incubated with ab279647 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse testis tissue labeling Lin28A with ab279647 at 1/100 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Positive staining on mouse undifferentiated spermatogonia (PMID: 32094118) is observed. The nuclear counterstain was DAPI (Blue).
Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)at 1/1000 dilution.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
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All lanes : Anti-Lin28A antibody [EPR24286-100] (ab279647) at 1/1000 dilution
Lane 1 : F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate
Lane 2 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 23 kDa
Observed band size: 25 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
The observed MW is consistent with what has been described in the literature (PMID:21962509)
Negative control: NIH/3T3 (PMID:21962509)
Exposure time: 3 seconds
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Immunohistochemical analysis of paraffin-embedded mouse ovary tissue labeling Lin28A with ab279647 at 1/1000 dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse oocytes (PMID: 23378032). The section was incubated with ab279647 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
-
Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling Lin28A with ab279647 at 1/1000 dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The section was incubated with ab279647 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Negative control: no staining on mouse liver (PMID: 11279525).
Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
-
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized F9 (mouse embryonal carcinoma epithelial cell line) cells labelling Lin28A with ab279647 at 1/500 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing nucleolar and cytoplasmic staining in F9 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).
Negative control: NIH/3T3 (PMID:21962509).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
-
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast, Left panel) / F9 (mouse embryonal carcinoma epithelial cell, Right panel) cells labeling Lin28A with ab279647 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Negative control: NIH/3T3 (PMID: 21962509).
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Lin28A was immunoprecipitated from 0.35 mg F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate 10 µg with ab279647 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab279647 at 1/5000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/10000 dilution.
Lane 1: F9 whole cell lysate 10 µg
Lane 2: ab279647 IP in F9 whole cell lysate 10 µg
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab279647 in F9 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 5.5 seconds
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Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling Lin28A with ab279647 at 1/1000 dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat undifferentiated spermatogonia. The section was incubated with ab279647 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (0)
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