Recombinant
RabMAb

Recombinant Anti-DGCR8 antibody [EPR18757] - BSA and Azide free (ab240325)

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Overview

  • Product name

    Anti-DGCR8 antibody [EPR18757] - BSA and Azide free
    See all DGCR8 primary antibodies

  • Description

    Rabbit monoclonal [EPR18757] to DGCR8 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, Flow Cyt, ICC/IF, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Human DGCR8 aa 400 to the C-terminus. The exact sequence is proprietary.
    Database link: Q8WYQ5

  • General notes

    Ab240325 is the carrier-free version of ab191875. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab240325 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

Properties

Applications

Our Abpromise guarantee covers the use of ab240325 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 Use at an assay dependent concentration. Detects a band of approximately 100 kDa (predicted molecular weight: 86 kDa).
Flow Cyt Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.

Target

  • Function

    Component of the microprocessor complex that acts as a RNA- and heme-binding protein that is involved in the initial step of microRNA (miRNA) biogenesis. Component of the microprocessor complex that is required to process primary miRNA transcripts (pri-miRNAs) to release precursor miRNA (pre-miRNA) in the nucleus. Within the microprocessor complex, DGCR8 function as a molecular anchor necessary for the recognition of pri-miRNA at dsRNA-ssRNA junction and directs DROSHA to cleave 11 bp away form the junction to release hairpin-shaped pre-miRNAs that are subsequently cut by the cytoplasmic DICER to generate mature miRNAs. The heme-bound DGCR8 dimer binds pri-miRNAs as a cooperative trimer (of dimers) and is active in triggering pri-miRNA cleavage, whereas the heme-free DGCR8 monomer binds pri-miRNAs as a dimer and is much less active. Both double-stranded and single-stranded regions of a pri-miRNA are required for its binding. Involved in the silencing of embryonic stem cells self-renewal.

  • Tissue specificity

    Ubiquitously expressed.

  • Sequence similarities

    Contains 2 DRBM (double-stranded RNA-binding) domains.
    Contains 1 WW domain.

  • Domain

    Both DRBM domains are required for efficient binding to pri-miRNA. The region between residues 276 and 498 has an autoinhibitory function on pri-miRNA processing activity.

  • Cellular localization

    Nucleus. Nucleus > nucleolus. Colocalizes with nucleolin and DROSHA in the nucleolus. Mostly detected in the nucleolus as electron-dense granular patches around the fibrillar center (FC) and granular component (GC). Also detected in the nucleoplasm as small foci adjacent to splicing speckles near the chromatin structure. Localized with DROSHA in GW bodies (GWBs), also known as P-bodies.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • DGCRK6 antibody
    • C22orf12 antibody
    • D16H22S788E antibody
    • D16Wis2 antibody
    • DGCR 8 antibody
    • Dgcr8 antibody
    • DGCR8 microprocessor complex subunit antibody
    • DGCR8_HUMAN antibody
    • DGCRK 6 antibody
    • DiGeorge syndrome critical region 8 antibody
    • DiGeorge syndrome critical region gene 8 antibody
    • Gy1 antibody
    • Microprocessor complex subunit DGCR8 antibody
    • pasha antibody
    see all

Images

  • Flow Cytometry - Anti-DGCR8 antibody [EPR18757] - BSA and Azide free (ab240325)
    Flow Cytometry - Anti-DGCR8 antibody [EPR18757] - BSA and Azide free (ab240325)

    Flow Cytometry analysis of Jurkat (human acute T cell leukemia) labelling DGCR8 with purified ab191875 at 1/1000 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Alexa Fluor® 488 goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

     

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab191875).

  • Immunocytochemistry/ Immunofluorescence - Anti-DGCR8 antibody [EPR18757] - BSA and Azide free (ab240325)
    Immunocytochemistry/ Immunofluorescence - Anti-DGCR8 antibody [EPR18757] - BSA and Azide free (ab240325)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling DGCR8 with ab191875 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing nuclear staining on HeLa cell line.

    The nuclear counterstain 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: ab191875 at 1/500 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab191875).

  • Immunocytochemistry/ Immunofluorescence - Anti-DGCR8 antibody [EPR18757] - BSA and Azide free (ab240325)
    Immunocytochemistry/ Immunofluorescence - Anti-DGCR8 antibody [EPR18757] - BSA and Azide free (ab240325)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling DGCR8 with ab191875 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing nuclear and weakly cytoplasmic staining on Jurkat cell line.

    The nuclear counterstain 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: ab191875 at 1/500 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab191875).

  • Immunoprecipitation - Anti-DGCR8 antibody [EPR18757] - BSA and Azide free (ab240325)
    Immunoprecipitation - Anti-DGCR8 antibody [EPR18757] - BSA and Azide free (ab240325)

    DGCR8 was immunoprecipitated from 1mg of HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate with ab191875 at 1/60 dilution.

    Western blot was performed from the immunoprecipitate using ab191875 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: HEK-293 whole cell lysate 10ug (Input).

    Lane 2: ab191875 IP in HEK-293 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab191875 in HEK-293 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 30 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab191875).

Protocols

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.

Click here to view the general protocols

References

Publishing research using ab240325? Please let us know so that we can cite the reference in this datasheet.

ab240325 has not yet been referenced specifically in any publications.

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