Recombinant Anti-Lin28B antibody [EPR18717] (ab191881)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18717] to Lin28B
- Suitable for: WB, ICC/IF, IP, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Human, Recombinant fragment
Related conjugates and formulations
Overview
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Product name
Anti-Lin28B antibody [EPR18717]
See all Lin28B primary antibodies -
Description
Rabbit monoclonal [EPR18717] to Lin28B -
Host species
Rabbit -
Tested applications
Suitable for: WB, ICC/IF, IP, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Human, Recombinant fragment -
Immunogen
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Recombinant full length Human Lin28B protein; Human placenta and testis lysates; K562, HepG2, HEK-293T and NCCIT whole cell lysates. ICC/IF: JAR and K562 cells. ICC/IF KO: HEK293 cells (HEK293-Lin28B KO used as a negative cell line) Flow Cyt (intra): K562 cells. IP: K562 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
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18717 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell pellets
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab191881 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
1/2000. Detects a band of approximately 30 kDa (predicted molecular weight: 27 kDa).
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ICC/IF |
Use a concentration of 1 - 10 µg/ml.
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IP |
1/50.
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Flow Cyt (Intra) |
1/150.
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Notes |
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WB
1/2000. Detects a band of approximately 30 kDa (predicted molecular weight: 27 kDa). |
ICC/IF
Use a concentration of 1 - 10 µg/ml. |
IP
1/50. |
Flow Cyt (Intra)
1/150. |
Target
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Function
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. 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. Mediates MYC-mediated let-7 repression. Isoform 1, when overexpressed, stimulates growth of the breast adenocarcinoma cell line MCF-7. Isoform 2 has no effect on cell growth. -
Tissue specificity
High expression in testis, fetal liver, placenta and in hepatocellular carcinoma (HCC). Isoform 1 is only detected in moderately and poorly differentiated HCC tissues and placenta (at protein level). Isoform 2 is detected in fetal liver, non-tumor liver tissues, as well as well-differentiated tumor tissues (at protein level). -
Sequence similarities
Belongs to the lin-28 family.
Contains 2 CCHC-type zinc fingers.
Contains 1 CSD (cold-shock) domain. -
Cellular localization
Cytoplasm. Nucleus. Predominantly cytoplasmic at G1 phase, accumulates in the nucleus in S and G2 phases. The frequency of nuclear localization in S and in G2 phases is 60% and 30%, respectively. - Information by UniProt
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Database links
- Entrez Gene: 389421 Human
- Omim: 611044 Human
- SwissProt: Q6ZN17 Human
- Unigene: 23616 Human
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Alternative names
- CSDD 2 antibody
- CSDD2 antibody
- FLJ16517 antibody
see all
Images
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All lanes : Anti-Lin28B antibody [EPR18717] (ab191881) at 1/2000 dilution
Lane 1 : Wild-type HEK293 cell lysate
Lane 2 : LIN28B knockout HEK293 cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : K562 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 27 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab191881 observed at 35 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab191881 was shown to react with Lin28B in wild-type HEK-293 cells in Western blot with loss of signal observed in LIN28B knockout sample. Wild-type HEK-293 and LIN28B knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab191881 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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ab191881 staining Lin28B in wild-type HEK293 cells (top panel) and Lin28B knockout HEK293 cells (bottom panel). The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab191881 at 10µg/ml and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a high-content analysis system (Perkin Elmer, Operetta CLS™).
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All lanes : Anti-Lin28B antibody [EPR18717] (ab191881) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : LIN28B knockout HEK293T cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : SW480 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 27 kDa
Observed band size: 36 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab191881 observed at 36 kDa. Red - loading control ab7291 observed at 50 kDa.
ab191881 Anti-Lin28B antibody [EPR18717] was shown to specifically react with Lin28B in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab265066 (knockout cell lysate ab257504) was used. Wild-type and Lin28B knockout samples were subjected to SDS-PAGE. ab191881 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Anti-Lin28B antibody [EPR18717] (ab191881) at 1/2000 dilution + Recombinant full length Human Lin28B protein at 0.01 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 27 kDa
Exposure time: 1 secondBlocking/Dilution buffer: 5% NFDM/TBST.
Recombinant Human Lin28B protein contains aa1-250 with His-Tag®.
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Anti-Lin28B antibody [EPR18717] (ab191881) at 1/2000 dilution + Human placenta lysate at 20 µg
Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 27 kDa
Observed band size: 30 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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Anti-Lin28B antibody [EPR18717] (ab191881) at 1/2000 dilution + Human testis lysate at 10 µg
Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 27 kDa
Observed band size: 30 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Lin28B antibody [EPR18717] (ab191881) at 1/2000 dilution
Lane 1 : K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate
Lane 2 : HepG2 (Human liver hepatocellular carcinoma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 27 kDa
Observed band size: 30 kDa why is the actual band size different from the predicted?
Exposure time: 5 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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Anti-Lin28B antibody [EPR18717] (ab191881) at 1/2000 dilution + NCCIT (Human pluripotent embryonic carcinoma) whole cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 27 kDa
Observed band size: 30 kDa why is the actual band size different from the predicted?
Exposure time: 15 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized JAR (Human placenta choriocarcinoma cell line) cells labeling Lin28B with ab191881 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasm and nuclear staining on JAR cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab191881 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution. -
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling Lin28B with ab191881 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasm and nuclear staining on K562 cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab191881 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution. -
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling Lin28B with ab191881 at 1/150 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.
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Lin28B was immunoprecipitated from 1mg of K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate with ab191881 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab191881 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.
Lane 1: K562 whole cell lysate 10ug (Input).
Lane 2: ab191881 IP in K562 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab191881 in K562 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 5 seconds.
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (6)
ab191881 has been referenced in 6 publications.
- Wang S et al. Transfer of microRNA-25 by colorectal cancer cell-derived extracellular vesicles facilitates colorectal cancer development and metastasis. Mol Ther Nucleic Acids 23:552-564 (2021). PubMed: 33510943
- Huang JH et al. Estrogen 17ß-estradiol accelerates the proliferation of uterine junctional zone smooth muscle cells via the let-7a/Lin28B axis in adenomyosis. Mol Med Rep 23:N/A (2021). PubMed: 34227673
- Mizushima E et al. Osteosarcoma-initiating cells show high aerobic glycolysis and attenuation of oxidative phosphorylation mediated by LIN28B. Cancer Sci 111:36-46 (2020). PubMed: 31705593
- Vishnubalaji R et al. Neoplastic Transformation of Human Mesenchymal Stromal Cells Mediated via LIN28B. Sci Rep 9:8101 (2019). PubMed: 31147574
- Wang X et al. RNA binding protein Lin28B confers gastric cancer cells stemness via directly binding to NRP-1. Biomed Pharmacother 104:383-389 (2018). PubMed: 29787985
- Zhao G et al. Placental exosome-mediated Bta-miR-499-Lin28B/let-7 axis regulates inflammatory bias during early pregnancy. Cell Death Dis 9:704 (2018). PubMed: 29899331