Rabbit Recombinant Monoclonal DGCR8 antibody. Carrier free. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples.
pH: 7.2 - 7.4
Constituents: PBS
IP | WB | ICC/IF | Flow Cyt (Intra) | |
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Human | Tested | Tested | Tested | Tested |
Mouse | Predicted | Expected | Predicted | Predicted |
Rat | Predicted | Expected | Predicted | Predicted |
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Species Human | Dilution info - | Notes - |
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Species Mouse, Rat | Dilution info - | Notes - |
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Species Human | Dilution info - | Notes - |
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Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
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Species Human | Dilution info - | Notes - |
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Species Mouse, Rat | Dilution info - | Notes - |
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Species Human | Dilution info - | Notes - |
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Species Mouse, Rat | Dilution info - | Notes - |
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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). Plays also a role in DNA repair by promoting the recruitment of RNF168 to RNF8 and MDC1 at DNA double-strand breaks and subsequently the clearance of DNA breaks (PubMed:34188037).
C22orf12, DGCRK6, LP4941, DGCR8, Microprocessor complex subunit DGCR8, DiGeorge syndrome critical region 8
Rabbit Recombinant Monoclonal DGCR8 antibody. Carrier free. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples.
pH: 7.2 - 7.4
Constituents: PBS
ab240325 is the carrier-free version of Anti-DGCR8 antibody [EPR18757] ab191875.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
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.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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.
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.
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.
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.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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Intracellular Flow Cytometry analysis of Jurkat (human acute T cell leukemia) labelling DGCR8 with purified Anti-DGCR8 antibody [EPR18757] 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 (Anti-DGCR8 antibody [EPR18757] ab191875).
DGCR8 was immunoprecipitated from 1mg of HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate with Anti-DGCR8 antibody [EPR18757] ab191875 at 1/60 dilution.
Western blot was performed from the immunoprecipitate using Anti-DGCR8 antibody [EPR18757] 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: Anti-DGCR8 antibody [EPR18757] ab191875 IP in HEK-293 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-DGCR8 antibody [EPR18757] 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 (Anti-DGCR8 antibody [EPR18757] ab191875).
All lanes: Immunoprecipitation - Anti-DGCR8 antibody [EPR18757] (Anti-DGCR8 antibody [EPR18757] ab191875)
Predicted band size: 86 kDa
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling DGCR8 with Anti-DGCR8 antibody [EPR18757] 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: Anti-DGCR8 antibody [EPR18757] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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 Anti-DGCR8 antibody [EPR18757] 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: Anti-DGCR8 antibody [EPR18757] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-DGCR8 antibody [EPR18757] ab191875).
This data was developed using Anti-DGCR8 antibody [EPR18757] ab191875, the same antibody clone in a different buffer formulation.
Western blot: Anti-DGCR8 antibody [EPR18757] (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, Anti-DGCR8 antibody [EPR18757] 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] (Anti-DGCR8 antibody [EPR18757] ab191875) at 1/1000 dilution
Lane 1: Wild-type HCT 116 cell lysate at 20 µg
Lane 2: Western blot - Human DGCR8 knockout HCT116 cell line (Human DGCR8 knockout HCT116 cell line ab287366)
Lane 2: DGCR8 knockout HCT 116 cell lysate at 20 µg
Lane 3: Wild-type A549 ab288558 cell lysate at 20 µg
Lane 4: DGCR8 knockout A549 Human DGCR8 knockout A549 cell line ab287368 cell lysate at 20 µg
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
Western blot: Anti-DGCR8 antibody [EPR18757] (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, Anti-DGCR8 antibody [EPR18757] 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] (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
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.
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