Recombinant
RabMAb

Recombinant Anti-Lin28B antibody [EPR18717] (ab191881)

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Overview

  • 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, Flow Cyt, IPmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant full length protein within Human Lin28B aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: Q6ZN17

  • Positive control

    • WB: Recombinant full length Human Lin28B protein; Human placenta and testis lysates; K562, HepG2 and NCCIT whole cell lysates. ICC/IF: JAR and K562 cells. Flow Cyt: K562 cells. IP: K562 whole cell lysate.

  • 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
    For more information see here.

    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

  • 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% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR18717

  • Isotype

    IgG

  • Research areas

Applications

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
WB 1/2000. Detects a band of approximately 30 kDa (predicted molecular weight: 27 kDa).
ICC/IF 1/1000.
Flow Cyt 1/150.
IP 1/50.

Target

  • 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

  • Database links

    • Alternative names

      • CSDD 2 antibody
      • CSDD2 antibody
      • FLJ16517 antibody
      • Lin 28 homolog B (C. elegans) antibody
      • Lin 28.2 antibody
      • Lin-28B antibody
      • LIN28B antibody
      • LN28B_HUMAN antibody
      • Protein lin-28 homolog B antibody
      see all

    Images

    • Western blot - Anti-Lin28B antibody [EPR18717] (ab191881)
      Western blot - Anti-Lin28B antibody [EPR18717] (ab191881)
      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 second


      Blocking/Dilution buffer: 5% NFDM/TBST.

      Recombinant Human Lin28B protein contains aa1-250 with His-Tag®.

    • Western blot - Anti-Lin28B antibody [EPR18717] (ab191881)
      Western blot - Anti-Lin28B antibody [EPR18717] (ab191881)
      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 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-Lin28B antibody [EPR18717] (ab191881)
      Western blot - Anti-Lin28B antibody [EPR18717] (ab191881)
      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 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-Lin28B antibody [EPR18717] (ab191881)
      Western blot - Anti-Lin28B antibody [EPR18717] (ab191881)
      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 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-Lin28B antibody [EPR18717] (ab191881)
      Western blot - Anti-Lin28B antibody [EPR18717] (ab191881)
      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 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunocytochemistry/ Immunofluorescence - Anti-Lin28B antibody [EPR18717] (ab191881)
      Immunocytochemistry/ Immunofluorescence - Anti-Lin28B antibody [EPR18717] (ab191881)

      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.

    • Immunocytochemistry/ Immunofluorescence - Anti-Lin28B antibody [EPR18717] (ab191881)
      Immunocytochemistry/ Immunofluorescence - Anti-Lin28B antibody [EPR18717] (ab191881)

      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.

    • Flow Cytometry - Anti-Lin28B antibody [EPR18717] (ab191881)
      Flow Cytometry - Anti-Lin28B antibody [EPR18717] (ab191881)

      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.

    • Immunoprecipitation - Anti-Lin28B antibody [EPR18717] (ab191881)
      Immunoprecipitation - Anti-Lin28B antibody [EPR18717] (ab191881)

      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.

    References

    This product has been referenced in:

    • Vishnubalaji R  et al. Neoplastic Transformation of Human Mesenchymal Stromal Cells Mediated via LIN28B. Sci Rep 9:8101 (2019). Read more (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). Read more (PubMed: 29787985) »
    See all 3 Publications for this product

    Publishing research using ab191881? Please let us know so that we can cite the reference in this datasheet.

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