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Rabbit Recombinant Monoclonal DGCR8 antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples. Cited in 35 publications.


Images

Western blot - Anti-DGCR8 antibody [EPR18757] (AB191875), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-DGCR8 antibody [EPR18757] (AB191875), expandable thumbnail
  • Western blot - Anti-DGCR8 antibody [EPR18757] (AB191875), expandable thumbnail
  • Western blot - Anti-DGCR8 antibody [EPR18757] (AB191875), expandable thumbnail
  • Western blot - Anti-DGCR8 antibody [EPR18757] (AB191875), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPWBICC/IFFlow Cyt (Intra)
Human
Tested
Tested
Tested
Tested
Mouse
Expected
Tested
Expected
Expected
Rat
Expected
Tested
Expected
Expected

Tested
Tested

Species

Human

Dilution info

1/60

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/1000

Notes

-

Species

Rat

Dilution info

1/1000

Notes

-

Species

Human

Dilution info

1/1000

Notes

-

Tested
Tested

Species

Human

Dilution info

1/1000

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

1/1000

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Associated Products

Select an associated product type

1 product for Alternative Product

Target data

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 (PubMed:26027739, PubMed:26748718). 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 (PubMed:15531877, PubMed:15574589, PubMed:15589161, PubMed:16751099, PubMed:16906129, PubMed:16963499, PubMed:17159994). Specifically recognizes and binds N6-methyladenosine (m6A)-containing pri-miRNAs, a modification required for pri-miRNAs processing (PubMed:25799998). Involved in the silencing of embryonic stem cell self-renewal (By similarity).

Alternative names

Microprocessor complex subunit DGCR8, DiGeorge syndrome critical region 8, DGCR8, C22orf12, LP4941, DGCRK6

Recommended products

Rabbit Recombinant Monoclonal DGCR8 antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples. Cited in 35 publications.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR18757

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

The DGCR8 protein also known as Pasha in some organisms serves an important role in the microRNA (miRNA) processing machinery. It has a molecular mass of approximately 90 kDa and shows expression ubiquitously but with higher levels in tissues with active cell division like the brain and lungs. DGCR8 acts as a partner in a complex with Drosha and together they initiate the microRNA maturation process by cleaving primary miRNA transcripts into precursor miRNA in the nucleus.

Biological function summary

DGCR8 partners with Drosha to form the Microprocessor complex an important player in miRNA biogenesis. This complex identifies and accurately processes primary transcripts into precursor miRNAs a step essential for proper gene regulation. DGCR8's role is critical for maintaining the normal cellular function enabling the precise control of miRNA production which in turn modulates gene expression post-transcriptionally. Aberrations in its function affect the regulation of genes involved in cell cycle differentiation and development.

Pathways

DGCR8 holds a significant influence on the RNA interference (RNAi) pathway interacting closely with Drosha to regulate miRNA synthesis. This pathway is fundamental for the regulation of genetic networks involved in cellular development and homeostasis. In the context of the miRNA processing pathway DGCR8 links to proteins like Dicer which further processes precursor miRNAs in the cytoplasm. Proper function of these pathways ensures the balanced expression of mRNAn important for healthy cellular activities.

Associated diseases and disorders

DGCR8 misregulation is associated with the development of DiGeorge syndrome and certain cancers. Alterations in its function can lead to compromised miRNA maturation impacting genes that control cell growth and survival. In DiGeorge syndrome haploinsufficiency of DGCR8 may contribute to the associated developmental anomalies. In cancer altered DGCR8 expression can disrupt miRNA profiles interacting further with proteins such as Argonaute to influence the dysregulation of gene expression profiles commonly observed in tumorigenesis.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

10 product images

  • Western blot - Anti-DGCR8 antibody [EPR18757] (ab191875), expandable thumbnail

    Western blot - Anti-DGCR8 antibody [EPR18757] (ab191875)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-DGCR8 antibody [EPR18757] (ab191875) at 1/1000 dilution

    Lane 1: HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate at 20 µg

    Lane 2: HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 20 µg

    Lane 3: WEHI-3 (Mouse leukemia cell line) whole cell lysate at 20 µg

    Lane 4: Neuro-2a (Mouse neuroblastoma cells) whole cell lysate at 20 µg

    Lane 5: Mouse testis lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution

    Predicted band size: 86 kDa

    Observed band size: 100 kDa

    Exposure time: 30s

  • Flow Cytometry (Intracellular) - Anti-DGCR8 antibody [EPR18757] (ab191875), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-DGCR8 antibody [EPR18757] (ab191875)

    Intracellular 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.

  • Western blot - Anti-DGCR8 antibody [EPR18757] (ab191875), expandable thumbnail

    Western blot - Anti-DGCR8 antibody [EPR18757] (ab191875)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-DGCR8 antibody [EPR18757] (ab191875) at 1/1000 dilution

    All lanes: Human fetal kidney lysate at 10 µg

    Secondary

    All lanes: Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/50000 dilution

    Predicted band size: 86 kDa

    Observed band size: 100 kDa

    Exposure time: 3min

  • Western blot - Anti-DGCR8 antibody [EPR18757] (ab191875), expandable thumbnail

    Western blot - Anti-DGCR8 antibody [EPR18757] (ab191875)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-DGCR8 antibody [EPR18757] (ab191875) at 1/1000 dilution

    Lane 1: Mouse brain lysate at 10 µg

    Lane 2: Rat brain lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution

    Predicted band size: 86 kDa

    Observed band size: 100 kDa

    Exposure time: 3min

  • Western blot - Anti-DGCR8 antibody [EPR18757] (ab191875), expandable thumbnail

    Western blot - Anti-DGCR8 antibody [EPR18757] (ab191875)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-DGCR8 antibody [EPR18757] (ab191875) at 1/1000 dilution

    Lane 1: PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 10 µg

    Lane 2: NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution

    Predicted band size: 86 kDa

    Observed band size: 100 kDa

    Exposure time: 5s

  • Immunoprecipitation - Anti-DGCR8 antibody [EPR18757] (ab191875), expandable thumbnail

    Immunoprecipitation - Anti-DGCR8 antibody [EPR18757] (ab191875)

    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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab191875 in HEK-293 whole cell lysate.

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

    Exposure time: 30 seconds.

    All lanes: Immunoprecipitation - Anti-DGCR8 antibody [EPR18757] (ab191875)

    Predicted band size: 86 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-DGCR8 antibody [EPR18757] (ab191875), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-DGCR8 antibody [EPR18757] (ab191875)

    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) (Goat Anti-Rabbit IgG H&L (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 Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed 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 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-DGCR8 antibody [EPR18757] (ab191875), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-DGCR8 antibody [EPR18757] (ab191875)

    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) (Goat Anti-Rabbit IgG H&L (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 Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed 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 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

  • Western blot - Anti-DGCR8 antibody [EPR18757] (ab191875), expandable thumbnail

    Western blot - Anti-DGCR8 antibody [EPR18757] (ab191875)

    Western blot: Anti-DGCR8 antibody [EPR18757] (ab191875) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab191875 was shown to bind specifically to DGCR8. A band was observed at 90-100 kDa in wild-type A549 cell lysates with no signal observed at this size in DGCR8 knockout cell line. To generate this image, wild-type and DGCR8 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

    All lanes: Western blot - Anti-DGCR8 antibody [EPR18757] (ab191875) at 1/1000 dilution

    Lane 1: Wild-type A549 cell lysate at 20 µg

    Lane 2: DGCR8 knockout A549 cell lysate at 20 µg

    Lane 3: HeLa cell lysate at 20 µg

    Lane 4: PC-3 cell lysate at 20 µg

    Secondary

    All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

    Performed under reducing conditions.

  • Western blot - Anti-DGCR8 antibody [EPR18757] (ab191875), expandable thumbnail

    Western blot - Anti-DGCR8 antibody [EPR18757] (ab191875)

    Western blot: Anti-DGCR8 antibody [EPR18757] (ab191875) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab191875 was shown to bind specifically to DGCR8. A band was observed at 100 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in DGCR8 knockout cell line. To generate this image, wild-type and DGCR8 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

    All lanes: Western blot - Anti-DGCR8 antibody [EPR18757] (ab191875) at 1/1000 dilution

    Lane 1: Wild-type HCT 116 cell lysate at 20 µg

    Lane 2: DGCR8 knockout HCT 116 cell lysate at 20 µg

    Lane 3: Wild-type A549 Human wild-type A549 cell line ab288558 cell lysate at 20 µg

    Lane 4: DGCR8 knockout A549 Human DGCR8 knockout A549 cell line ab287368 cell lysate at 20 µg

    Secondary

    Lanes 1 - 4: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

    Lanes 1 - 4: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

    Performed under reducing conditions.

    Observed band size: 100 kDa

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