Anti-Lin28A antibody [EPR24286-100] (ab279647)
![Anti-Lin28A antibody [EPR24286-100] (ab279647)](https://www.abcam.com/ps/products/279/ab279647/Images/ab279647-451605-anti-lin28a-antibody-epr24286100-western-blot-embryonic.jpg "Anti-Lin28A antibody [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, ICC, IP, IHC-P, IHC-Fr
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-Lin28A antibody [EPR24286-100]
See all Lin28A primary antibodies -
Description
Rabbit monoclonal [EPR24286-100] to Lin28A -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt MouseHumanICC MouseHumanIHC-Fr MouseIHC-P MouseRatIP MouseHumanWB MouseHuman
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Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: NCCIT, JAR and F9 whole cell lysate. IHC-P: Mouse testis and ovary tissue. Rat testis tissue. IHC-Fr: Mouse testis tissue. ICC: F9 and NCCIT cells. Flow Cyt: F9 and NCCIT cells. IP: F9 and NCCIT whole cell lysate.
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General notes
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.
Properties
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Form
Liquid -
Storage instructions
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. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR24286-100 -
Isotype
IgG -
Research areas
Images
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Western blot - Anti-Lin28A antibody (ab279647)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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Immunocytochemistry - Anti-Lin28A antibody (ab279647)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 Cytometry - Anti-Lin28A antibody (ab279647)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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Immunoprecipitation - Anti-Lin28A antibody (ab279647)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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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lin28A antibody (ab279647)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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Immunohistochemistry (Frozen sections) - Anti-Lin28A antibody (ab279647)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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Western blot - Anti-Lin28A antibody (ab279647)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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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lin28A antibody (ab279647)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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lin28A antibody (ab279647)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.
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Immunocytochemistry - Anti-Lin28A antibody (ab279647)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.
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Flow Cytometry - Anti-Lin28A antibody (ab279647)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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Immunoprecipitation - Anti-Lin28A antibody (ab279647)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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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lin28A antibody (ab279647)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.