DGCR8 / Microprocessor complex subunit DGCR8 · IHC design guide

Design Immunohistochemistry for DGCR8

Plan paraffin-section DGCR8 IHC using general nuclear staining as the tissue benchmark (HPA tissue IHC). Compare high-staining glandular cells with cell populations reported as low or undetected, using the IHC-validated antibody's documented conditions (HPA tissue IHC; datasheet A00475-1).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use

Immunohistochemistry protocol sheet for DGCR8 (IHC for DGCR8): expected localisation General nuclear staining (HPA tissue IHC), antibody A00475-1, validated IHC image, and IHC protocol steps — Printable **DGCR8** IHC protocol sheet — expected localisation General nuclear staining (HPA tissue IHC), antibody A00475-1, controls and protocol steps. Open the full DGCR8 IHC guide →

Navigate to: Protocols Staining pattern Controls Antibodies

DGCR8 Immunohistochemistry Experimental Design Guide

Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.

Must know before staining

	Expected localisation	General nuclear staining (HPA tissue IHC)
	Staining pattern	Nuclear staining across diverse tissue cell types (HPA tissue IHC)
	Antigen retrieval	Citrate pH 6 HIER, heat-mediated (datasheet A00475-1)
	Positive control ⓘ	Appendix+4 more · see all
	Negative control ⓘ	Adipose tissue+4 more · see all

Important caveats

Reasons your staining may differ from the expected pattern.

	Fixation	Keep fixation consistent across sections. (standard IHC practice; not target-specific)
	Caveat	Staining varies by cell type; adipocytes may be negative (HPA tissue IHC)
	Regulation	Ubiquitously expressed (UniProt)
	Isoform / epitope	3 isoforms; map the antibody epitope to each sequence (UniProt)

Section 1

Recommended DGCR8 IHC & IF Protocols

Compare the catalog antibody’s IHC-P protocol with published DGCR8 staining of testis, breast carcinoma, and melanoma sections (PMC7237130; PMC8242032; PMC12797089).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded human rectal cancer tissue; fixative not specified (datasheet A00475-1)
Fixation	Image fixative and duration unreported (datasheet A00475-1); verify before use.
Sectioning	4–5 µm sections on charged slides (standard)
Deparaffinisation	Xylene, graded ethanol series to water (standard)
Antigen retrieval	Heat retrieval: Citrate pH 6, 20 min (datasheet A00475-1)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% goat serum (datasheet A00475-1)
Primary antibody	Rabbit anti-DGCR8, 0.5-1μg/ml (datasheet A00475-1)
Primary incubation	Overnight at 4 °C (datasheet A00475-1)
Detection	Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A00475-1)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	DGCR8-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: General nuclear expression. No signal in the no-primary control.

💡Decision noteStart with heat-mediated citrate retrieval at pH 6 (datasheet: A00475-1); the testis protocol also reports citrate retrieval at pH 6.0 (PMC7237130).

Section 2

What Is the Expected DGCR8 Staining Pattern?

DGCR8 is a nuclear protein with no transmembrane segment (UniProt Q8WYQ5 topology). In paraffin-section IHC, expect predominantly nuclear staining, including in glandular cells of the colon and appendix and trophoblastic cells of the placenta (HPA: general nuclear expression; High in these cells). HPA rates its tissue IHC profile Approved, with medium consistency between antibody staining and RNA expression; interpret intensity with that limitation (HPA: reliability).

What am I looking at on my slide?
Distinct nuclear staining in colon or appendix glandular cells, with relatively clear surrounding tissue.	This fits the reported high staining in those cells and the general nuclear IHC pattern (HPA: High in colon and appendix glandular cells; general nuclear expression). Score the nuclear signal in the named cells. Nucleolar detail may be difficult to resolve with chromogenic detection, even though DGCR8 has been detected there (UniProt Q8WYQ5 localization; standard IHC practice).
Strong staining is predominantly cytoplasmic or membranous while nuclei are faint.	This is the wrong dominant compartment for the reported tissue pattern (HPA: general nuclear expression). DGCR8 has no transmembrane segment (UniProt Q8WYQ5 topology). Treat the result as suspect and compare it with a known-positive nuclear pattern before assigning the signal to DGCR8 (standard IHC interpretation practice).
A cell type reported as undetected stains strongly, such as smooth muscle cells or adipocytes.	HPA reports no detected staining in smooth muscle cells and adipocytes in the listed tissues (HPA: Not detected). A strong result there warrants checks for antibody cross-reactivity or endogenous detection activity (standard IHC practice). It does not, by itself, overturn the reported pattern; assess it alongside nuclear localization and controls.
Weak color spreads across nuclei, cytoplasm, and extracellular areas with little cell-to-cell distinction.	Diffuse staining without nuclear enrichment is hard to reconcile with DGCR8's reported localization (HPA: general nuclear expression; UniProt Q8WYQ5 localization). Check background with a no-primary control, then review blocking, washes, detection, and counterstain as general IHC variables (standard IHC practice).
No nuclear signal appears in an adequately preserved colon glandular-cell section.	That conflicts with the reported high staining in colon glandular cells (HPA: High in colon glandular cells). First verify the positive-control section and detection run; then review the catalog antibody's validated IHC-P instructions and the run's retrieval and dilution settings (standard IHC practice). The supplied sources do not establish DGCR8-specific fixation sensitivity.

💡Expected DGCR8 appearanceCall a positive result when nuclei of expected cells show clear, preferably strong staining—such as colon glandular cells (HPA: High)—and treat dominant cytoplasmic, membranous, or diffuse staining as suspect (HPA: general nuclear expression; UniProt Q8WYQ5 topology; standard IHC interpretation practice).

How each factor affects the staining
Choice of comparison tissue and cell type	Colon, appendix, duodenum, gallbladder, rectum, and salivary-gland glandular cells are reported High; placenta trophoblastic cells and testis round or early spermatids are also High (HPA: tissue IHC). Judge a run against its specific expected cell type, since HPA also lists Low and Not detected cell populations in other tissues (HPA: tissue IHC).
Subcellular resolution and application	UniProt describes nucleolar patches and small nucleoplasmic foci (UniProt Q8WYQ5 localization). HPA tissue IHC summarizes general nuclear expression, while its ICC-IF data support nucleoplasm and nuclear bodies (HPA: tissue IHC; ICC-IF). Use those finer locations as context, without requiring individual puncta to be visible in chromogenic paraffin sections (standard IHC practice).
Antibody validation and isoform coverage	The tissue profile is Approved with medium RNA-to-staining consistency, and HPA079351 is IHC Approved (HPA: reliability; antibody validation). DGCR8 has 3 listed isoforms (UniProt Q8WYQ5 isoforms). The supplied evidence gives no epitope location or isoform-specific staining comparison, so do not infer that an IHC result detects every isoform.
Endogenous detection activity	A no-primary control can reveal color generated by the detection system rather than primary-antibody binding (standard chromogenic IHC practice). This check matters when color appears in HPA-undetected cell types or outside nuclei, but it cannot alone identify antibody cross-reactivity (HPA: Not detected cell types; general nuclear expression; standard IHC practice).

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Expected-positive glandular cells have no visible nuclear staining.	The run may have a detection or setup problem; HPA reports High staining in colon glandular cells (HPA: tissue IHC; standard IHC practice).	Check the positive-control section and detection reagents, then follow the catalog antibody's validated IHC-P instructions for retrieval and dilution (standard IHC practice). Do not assume a DGCR8-specific fixation effect from the supplied evidence.
Brown signal is strong in cytoplasm or at cell borders.	The dominant location disagrees with general nuclear tissue staining and DGCR8's lack of a transmembrane segment (HPA: tissue IHC; UniProt Q8WYQ5 topology).	Compare with a known-positive nuclear pattern and a no-primary control; review blocking and detection background before scoring these cells positive (standard IHC practice).
Smooth muscle cells or adipocytes stain as strongly as the intended positive cells.	HPA lists these cell types as Not detected in the specified tissues; cross-reactivity or endogenous detection activity is possible (HPA: tissue IHC; standard IHC practice).	Check cell identity and localization, inspect the no-primary control, and interpret the result against the positive-control tissue (standard IHC practice). Avoid treating one unexpected cell population as proof of DGCR8 expression.
The whole section has a weak, even haze.	Broad background can obscure nuclear enrichment and may reflect general staining or detection conditions (standard IHC practice).	Inspect the no-primary control and review blocking, washes, detection, and counterstain (standard IHC practice). Reassess whether nuclei of the HPA-listed positive cells remain distinguishable (HPA: tissue IHC).
One section stains while another tissue or cell population appears negative.	HPA reports High, Low, and Not detected populations despite low RNA tissue specificity (HPA: tissue IHC; RNA specificity).	Compare like cell types and use a reported High population as the run control (HPA: tissue IHC; standard IHC practice). Do not classify every negative population as an assay failure.
Does an IF/ICC image with nucleoplasmic and nuclear-body signal fit DGCR8?	HPA supports both ICC-IF locations; UniProt also reports nucleolar and nucleoplasmic localization (HPA: ICC-IF; UniProt Q8WYQ5 localization).	Yes—interpret that image within its IF/ICC context (HPA: ICC-IF). For paraffin IHC, use the general nuclear tissue pattern to score staining (HPA: tissue IHC); IF/ICC methods belong in their separate guide.

Source: UniProt Q8WYQ5 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing DGCR8 staining

IHC analysis of DGCR8 using anti-DGCR8 antibody (A00475-1). DGCR8 was detected in paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-DGCR8 Antibody (A00475-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

A00475-1 · IHCIHC analysis of DGCR8 using anti-DGCR8 antibody (A00475-1). DGCR8 was detected in paraffin-embedded section of human rectal cancer tissue. H

IF analysis of DGCR8 using anti-DGCR8 antibody (A00475-1). DGCR8 was detected in immunocytochemical section of A431 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-DGCR8 Antibody (A00475-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

A00475-1 · IFIF analysis of DGCR8 using anti-DGCR8 antibody (A00475-1). DGCR8 was detected in immunocytochemical section of A431 cell. Enzyme antigen ret

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000128191-DGCR8/tissue/cerebral+cortex#imid_21036395

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000128191-DGCR8/tissue/colon#imid_21039417

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000128191-DGCR8/tissue/liver#imid_21039418

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000128191-DGCR8/tissue/kidney#imid_21039411

HPA · ICC-IF REHHuman Protein Atlas immunofluorescence, REH, antibody HPA076916; green = DGCR8, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

Real IHC / IF data behind the staining-pattern expectations in this section.

Sample controls for DGCR8 IHC & IF

🧪Run colon first: glandular cells should stain (HPA: High in colon glandular cells). Use adipose tissue adipocytes as the biological low-signal comparison (HPA: Not detected in adipocytes); on the colon slide, unstained non-glandular cells can show local background, but their DGCR8 status is not established by these rows.

Positive control tissue: Appendix (Glandular cells, HPA High)

Negative control tissue: Adipose tissue (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show DGCR8 in REH, SiHa, U2OS, A-431, K-562, with annotated localisation: Nucleoplasm (supported), Nuclear bodies (supported) (HPA subcellular).

Technical controls: Include a no-primary (secondary-only) slide, a concentration-matched rabbit IgG isotype control matched to the primary antibody’s clonality where applicable (caption: rabbit anti-DGCR8), and a DGCR8 knockout specimen or validated peptide-block control. For colon, quench endogenous peroxidase and check for endogenous biotin background before using the caption’s biotin/SABC-DAB detection (caption: biotinylated secondary, SABC and DAB).

⚠️Feasibility: A target-specific fixation window and fixation effect are unreported; the selected A00475-1 paraffin-section caption does not state the fixative (caption: fixative unreported). The reported IHC procedure uses heat retrieval in citrate buffer at pH 6 for 20 minutes, followed by 1 μg/ml primary antibody overnight at 4°C (caption: A00475-1 tissue IHC). No matched evidence establishes whether frozen sections or IF are easier; ICC-IF images support nucleoplasmic and nuclear-body localization (HPA: supported subcellular locations), while the biotin/SABC-DAB workflow warrants a background check in colon (caption: biotin/SABC-DAB).

HPA tissue IHC evidence for DGCR8

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.

Positive expression · recommended positive controls

Tissue	Cell type	Level	Evidence	Source
Appendix	Glandular cells	High	Protein (IHC)	HPA →
Colon	Glandular cells	High	Protein (IHC)	HPA →
Duodenum	Glandular cells	High	Protein (IHC)	HPA →
Gallbladder	Glandular cells	High	Protein (IHC)	HPA →
Placenta	Trophoblastic cells	High	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Adipose tissue	Adipocytes	Not detected	Protein (IHC)	HPA →
Adrenal gland	Glandular cells	Not detected	Protein (IHC)	HPA →
Breast	Adipocytes	Not detected	Protein (IHC)	HPA →
Liver	Cholangiocytes	Not detected	Protein (IHC)	HPA →
Ovary	Follicle cells	Not detected	Protein (IHC)	HPA →

Section 3

Advanced DGCR8 IHC Tips

Troubleshoot DGCR8 staining in paraffin sections by checking nuclear localisation, tissue controls and the documented conditions for the IHC-validated antibody.

How should I adjust retrieval when DGCR8 nuclear staining is weak?

Start with heat-mediated retrieval in citrate buffer at pH 6 for 20 minutes (datasheet A00475-1). Compare a strong expected compartment, such as rectal glandular cells, with a matched no-primary control before increasing retrieval intensity (HPA: High in rectal glandular cells; standard IHC practice). If staining remains weak, test a longer heating interval on adjacent sections while keeping antibody concentration and detection constant (standard IHC practice). Assess tissue preservation alongside nuclear signal, since excessive heating can damage morphology and complicate scoring (standard IHC practice). Record whether signal appears in nuclei rather than diffuse cytoplasm (HPA: general nuclear expression).

Could fixation explain weak DGCR8 staining in paraffin sections?

Target-specific fixation sensitivity is unknown because the selected antibody’s paraffin-section caption does not state a fixative (datasheet A00475-1). Record the actual fixative and fixation duration for each specimen, then compare sections processed together under the same retrieval and staining conditions (standard IHC practice). Use the documented citrate pH 6, 20-minute retrieval as the starting condition, and judge any change against preserved morphology and nuclear contrast (datasheet A00475-1; standard IHC practice). Do not assign a DGCR8-specific fixation effect from staining patterns alone; compare technical replicates and a no-primary control before interpreting weak signal (standard IHC practice).

Is cytoplasmic or punctate staining consistent with DGCR8?

Prioritise nuclear staining in paraffin sections, because tissue IHC shows general nuclear expression and cellular imaging supports nucleoplasm and nuclear bodies (HPA: tissue IHC; HPA: subcellular). DGCR8 has also been described in nucleoli and small nucleoplasmic foci, so nuclear puncta can be biologically plausible (UniProt Q8WYQ5: subcellular location). Localisation with DROSHA in GW bodies has been reported, but diffuse cytoplasmic DAB should still prompt checks of background and morphology before interpretation (UniProt Q8WYQ5: subcellular location; standard IHC practice). Compare suspect regions with adjacent intact cells and a no-primary section, especially where pigment or damaged tissue obscures the nuclear boundary (standard IHC practice).

Can isoforms or epitope accessibility explain inconsistent DGCR8 IHC?

DGCR8 has 3 annotated isoforms, while the supplied antibody caption does not identify its epitope or establish isoform coverage (UniProt Q8WYQ5: isoforms; datasheet A00475-1). Do not treat differences between sections as isoform-specific staining without an epitope map and independent validation (standard IHC practice). Its WW and double-stranded RNA-binding domains, together with annotated phosphorylation sites, make epitope position worth checking if that information becomes available (UniProt Q8WYQ5: domains and modified residues). For the current antibody, compare adjacent sections using identical citrate pH 6 retrieval and detection, then assess nuclear signal against morphology and controls (datasheet A00475-1; standard IHC practice).

How can IF help resolve ambiguous DGCR8 IHC localisation?

Use IF as a separate localisation check when chromogenic boundaries are ambiguous, assessing DGCR8 against a nuclear counterstain and a marker for the expected cell population (standard IF practice). For rectal glandular cells, select an epithelial marker and inspect whether DGCR8 signal occupies nuclei within that population (HPA: High in rectal glandular cells; HPA: general nuclear expression). Choose fluorophores after checking the specimen’s autofluorescence in each channel, and include single-label controls when signals overlap (standard IF practice). DGCR8 lacks a transmembrane segment and is mainly nuclear, so permeabilisation should allow antibody access to nuclear epitopes without assuming that the unknown antibody epitope is exposed (UniProt Q8WYQ5: topology and localisation; standard IF practice).

What should I check when DGCR8 DAB staining is widespread?

The documented paraffin-section example used 10% goat serum blocking, 1 µg/ml primary antibody overnight at 4 °C, and biotin-based DAB detection (datasheet A00475-1). Run a no-primary section and inspect endogenous pigment and staining outside intact nuclei before calling widespread brown signal DGCR8 (standard IHC practice; HPA: general nuclear expression). A peroxidase block is a general chromogenic IHC control, while a detection-system control can help identify background associated with the biotin-based workflow (standard IHC practice; datasheet A00475-1). If background persists, optimise primary concentration and washing on matched sections while retaining a tissue region with expected nuclear staining (standard IHC practice; HPA: High in rectal glandular cells).

How should I quantify DGCR8 across heterogeneous tissue regions? ⚠ ANSWER MARKED FOR VERIFICATION

Score intact nuclei within a defined cell population, recording both the percentage positive and staining intensity; an H-score can summarise their combination on a 0–300 scale (HPA: general nuclear expression; standard IHC practice). Normalise each result to the number of evaluable nuclei in that population, rather than total tissue area when cellularity varies (standard IHC practice). If reporting positive-cell density per mm², also report cell density so sparse and crowded regions can be compared sensibly (standard IHC practice). Keep retrieval, imaging, thresholding and region-selection rules constant, and exclude necrotic or folded areas before comparing specimens (standard IHC practice).

How do I distinguish true DGCR8 positivity from artefact?

A credible positive pattern is predominantly nuclear in intact cells, consistent with the tissue profile and supported nucleoplasmic or nuclear-body localisation (HPA: tissue IHC; HPA: subcellular). Rectal glandular cells offer a high-expression reference, whereas adipocytes in adipose tissue are reported as not detected; interpret each compartment on its own terms (HPA: High in rectal glandular cells; HPA: Not detected in adipocytes). Treat staining confined to section edges, necrotic areas or diffuse cytoplasm as suspect until it survives matched controls and morphology review (standard IHC practice). Check a no-primary section for endogenous enzyme or pigment signal before attributing brown deposits to DGCR8 (standard IHC practice).

Boster reagents

Best DGCR8 / Microprocessor complex subunit DGCR8 IHC Antibodies

A00475-1 has IHC images from human rectal cancer and mouse small intestine paraffin sections, plus IF in A431 cells (catalog image captions); both SKUs list human, mouse and rat reactivity (catalog).

Real IHC data

Anti-DGCR8 Antibody ®

Cat # A00475-1

View product →

A00475-1 has IHC images from human rectal cancer and mouse small intestine paraffin sections, and an IF image from A431 cells (catalog image captions). M00475 lists ICC/IF among its applications and human, mouse and rat reactivity, but has no supplied IHC or IF image (catalog).

Which to pick: For tissue IHC, choose A00475-1: its paraffin-section captions document citrate retrieval for 20 minutes and 1 μg/ml primary antibody; the fixative is unreported (A00475-1 IHC image captions). For IF/ICC, A00475-1 has an A431 IF image at 2 μg/ml, while M00475 is a monoclonal ICC/IF option without a supplied image (catalog IF image caption; catalog applications and clone). Both list human, mouse and rat reactivity, but the supplied tissue IHC images show only human and mouse samples (catalog reactivity; A00475-1 IHC image captions).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

UniProt Consortium. UniProt entry Q8WYQ5 (DGCR8_HUMAN, Microprocessor complex subunit DGCR8).
Human Protein Atlas. DGCR8 tissue IHC expression (reliability: Approved).
Human Protein Atlas. DGCR8 subcellular location (ICC-IF): Mainly localized to the nucleoplasm and nuclear bodies..
Human Protein Atlas. DGCR8 antibody validation summary (2 antibodies).
Deficient Expression of DGCR8 in Human Testis is Related to Spermatogenesis Dysfunction, Especially in Meiosis I. International journal of general medicine 2020 — PMC7237130.
CircKPNB1 mediates a positive feedback loop and promotes the malignant phenotypes of GSCs via TNF-α/NF-κB signaling. Cell death & disease 2022 — PMC9363451.
Non-canonical function of DGCR8 in DNA double-strand break repair signaling and tumor radioresistance. Nature communications 2021 — PMC8242032.
Risk Factors for Melanoma Survival: DGCR8 as a Predictive Factor for Mortality in Young Patients. Acta dermato-venereologica 2026 — PMC12797089.
PubMed PMID:12705904 — UniProt-cited evidence.
PubMed PMID:15461802 — UniProt-cited evidence.
PubMed PMID:14702039 — UniProt-cited evidence.