DGCR8 / Microprocessor complex subunit DGCR8 · IHC design guide

Design Immunohistochemistry for DGCR8

Plan DGCR8 paraffin IHC around its general nuclear tissue pattern (HPA tissue IHC). Use colon glandular cells as a high-staining reference and adipocytes as a low-staining comparison, then score nuclear signal with the IHC-validated antibody (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 in tissue (HPA tissue IHC), antibody A00475-1, validated IHC image, and IHC protocol steps — Printable **DGCR8** IHC protocol sheet — expected localisation General nuclear staining in tissue (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 in tissue (HPA tissue IHC)
	Staining pattern	General nuclear staining across tissues (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 paraffin sections. (standard IHC practice; not target-specific)
	Caveat	Staining has medium consistency with RNA data (HPA tissue IHC)
	Regulation	Ubiquitously expressed (UniProt)
	Isoform / epitope	3 isoforms; epitope coverage is unspecified (UniProt)

Section 1

Recommended DGCR8 IHC & IF Protocols

The catalog antibody’s IHC-P protocol (datasheet A00475-1) is accompanied by 4 published DGCR8 IHC protocols (PMC7237130; PMC9363451; 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 citrate pH 6 heat retrieval (datasheet A00475-1; PMC7237130), then optimize antibody dilution for the tissue (standard IHC practice).

Section 2

What Is the Expected DGCR8 Staining Pattern?

DGCR8 should appear chiefly in nuclei, including nucleoplasm and nucleoli (UniProt Q8WYQ5; HPA subcellular: supported nucleoplasm and nuclear bodies). Expect staining in several glandular cell populations and placental trophoblastic cells (HPA tissue IHC: High). DGCR8 has no transmembrane segment (UniProt Q8WYQ5 topology). HPA rates its tissue IHC profile Approved, with medium consistency between staining and RNA expression (HPA tissue IHC).

What am I looking at on my slide?
Distinct nuclear staining in colonic or duodenal glandular cells.	This fits the reported High staining in those cells (HPA tissue IHC). Score the nuclear signal in the named cell population; do not treat every cell in the section as an equivalent positive control (general IHC practice).
Predominantly cytoplasmic or membranous staining, with little nuclear signal.	This conflicts with DGCR8’s nuclear localization (UniProt Q8WYQ5; HPA subcellular: supported nucleoplasm and nuclear bodies). Treat it as a possible staining artefact and review controls and detection conditions before assigning it to DGCR8 (general IHC practice).
Strong signal in adipocytes or smooth muscle cells.	HPA reports these cells as Not detected in its sampled tissues (HPA tissue IHC). Check for cross-reactivity or endogenous detection activity (general IHC practice). The HPA result is an observed pattern, not proof that every such cell must be negative.
Diffuse colour across cells and surrounding tissue obscures nuclear detail.	The result cannot be scored reliably against the reported general nuclear pattern (HPA tissue IHC). Review background in a no-primary control, then assess blocking, antibody concentration and detection development (general IHC practice).
No nuclear signal in appendix or colon glandular cells.	These are reported High in HPA tissue IHC, so a blank section warrants a technical check. Verify tissue identity and examine a positive control before concluding that the sample lacks DGCR8 (general IHC practice).

💡Expected DGCR8 appearanceA convincing IHC positive shows clear nuclear staining in glandular cells of appendix or colon, where HPA reports High staining; dominant membrane or cytoplasmic colour without nuclear detail is suspect (HPA tissue IHC; UniProt Q8WYQ5 localization).

How each factor affects the staining
Nuclear distribution	UniProt reports nucleolar patches and small nucleoplasmic foci; HPA supports nucleoplasm and nuclear bodies by ICC-IF (UniProt Q8WYQ5; HPA subcellular). Fine detail may be harder to resolve in chromogenic tissue IHC (general IHC practice).
Cell population	HPA reports High staining in several glandular populations, trophoblastic cells and round or early spermatids, but Not detected in selected other populations (HPA tissue IHC). Interpret each cell type separately.
IHC antibody evidence	HPA079351 is IHC Approved; HPA076916 has no IHC status listed (HPA antibody validation). HPA’s tissue profile has medium staining–RNA consistency, so an unexpected pattern needs controls rather than an automatic biological explanation (HPA tissue IHC).
Isoforms and epitope	UniProt lists 3 isoforms (UniProt Q8WYQ5). The supplied evidence does not map an IHC antibody epitope to them; check the chosen antibody’s documentation before making isoform-specific claims (general IHC practice).
Processing and topology	UniProt lists one chain, residues 1–773, with no signal peptide, propeptide or transmembrane segment (UniProt Q8WYQ5). These annotations support a nuclear interpretation; they provide no basis for a shed extracellular staining pattern.
IF/ICC Q: What should I expect?	A: Mainly nucleoplasm and nuclear bodies, both supported by HPA ICC-IF; UniProt also reports nucleolar localization (HPA subcellular; UniProt Q8WYQ5). This IF/ICC expectation does not establish a chromogenic IHC staining intensity.
Fixation sensitivity	Target-specific fixation sensitivity is unreported in the supplied UniProt and HPA evidence. Record fixation and retrieval conditions when comparing sections, without attributing a change in DGCR8 signal to fixation alone (general IHC practice).

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Known-positive glandular tissue is blank.	The result disagrees with reported High staining in appendix or colon glandular cells (HPA tissue IHC); tissue selection or an IHC step may have failed (general IHC practice).	Confirm the sampled cell population and run a positive tissue control; review retrieval, antibody dilution and detection settings against the chosen IHC workflow (general IHC practice).
Colour is mostly cytoplasmic.	That pattern is discordant with the reported nuclear localization (UniProt Q8WYQ5; HPA tissue IHC). Nonspecific staining or detection background is possible (general IHC practice).	Compare with a no-primary control and reassess antibody concentration, blocking and development time before scoring DGCR8 positivity (general IHC practice).
Adipocytes or smooth muscle stain strongly.	HPA lists those cell types as Not detected in its tissue observations (HPA tissue IHC). Cross-reactivity or endogenous detection activity should be considered (general IHC practice).	Inspect no-primary and detection-only controls; assess whether colour follows the cell type or appears broadly across the section (general IHC practice).
Weak nuclear colour is hard to distinguish from background.	Diffuse background can obscure the reported general nuclear pattern (HPA tissue IHC); excess detection colour or insufficient blocking can reduce contrast (general IHC practice).	Compare controls, then adjust blocking, antibody concentration or chromogen development within the chosen IHC workflow (general IHC practice).
Two tissues show different proportions of positive cells.	HPA reports staining by cell population, with levels from High to Not detected across its sampled tissues (HPA tissue IHC). Different cell composition can affect the section-wide impression.	Score comparable cell populations and record intensity and proportion separately; include tissue identity in the interpretation (general IHC practice).
The two listed HPA antibodies appear to support different applications.	HPA079351 is IHC Approved and ICC Supported; HPA076916 is ICC Supported with no IHC status listed (HPA antibody validation). ICC support alone does not establish IHC performance.	For paraffin-section IHC, use an IHC-validated antibody and judge its nuclear pattern against the tissue controls (HPA antibody validation; general IHC practice).

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: its glandular cells must show DGCR8 staining (HPA: High in colon glandular cells). Use adipose tissue adipocytes as the negative tissue (HPA: Not detected in adipocytes); on the colon slide, treat any unstained non-glandular cells as an internal background reference, without assuming they are DGCR8-negative.

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 control; an isotype control matched to the rabbit primary’s clonality, which the caption does not report (tissue-IHC caption: rabbit primary); and, where available, DGCR8 knockout tissue or a validated peptide block as a biological specificity control. Quench endogenous peroxidase and check endogenous biotin background when using the caption’s biotin-based DAB detection in colon (tissue-IHC caption: biotinylated secondary, SABC and DAB).

⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected A00475-1 tissue-IHC caption does not state a fixative (selected tissue-IHC caption: fixative not stated). The paraffin-section example used heat retrieval in citrate buffer at pH 6 for 20 minutes; it does not establish whether retrieval is required under other conditions (selected tissue-IHC caption). Relative ease of frozen-section staining versus IF is unreported; for colon, assess background from endogenous peroxidase or biotin with the caption’s SABC/DAB method (selected tissue-IHC caption: 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 retrieval, nuclear localisation and cell-specific controls before interpreting signal intensity.

What retrieval should I try when DGCR8 nuclear staining is weak?

Start with heat-mediated citrate retrieval at pH 6 for 20 minutes (datasheet A00475-1). The selected paraffin-section image used that treatment before overnight incubation with 1 μg/mL catalog antibody at 4°C (datasheet A00475-1). If staining remains weak, compare retrieval duration on adjacent sections while holding antibody concentration, development time and imaging conditions constant (standard IHC practice). Excessive retrieval can damage morphology or raise background, so assess nuclear detail and an appropriate positive control alongside signal intensity (standard IHC practice). A weak result alone cannot establish loss of DGCR8 expression (standard IHC interpretation).

Could fixation explain weak DGCR8 staining in my paraffin sections?

Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state a fixative (datasheet A00475-1). Record the fixative and fixation duration for each specimen, then compare sections processed with the same retrieval, antibody incubation and chromogen development (standard IHC practice). The demonstrated workflow used citrate at pH 6 for 20 minutes and 1 μg/mL antibody overnight at 4°C (datasheet A00475-1). If similarly processed positive-control sections stain while a test section does not, review specimen handling and tissue preservation before assigning a biological explanation (standard IHC practice). Nuclear morphology can help identify compromised sections (standard histology practice).

Should DGCR8 stain nucleoli, nucleoplasm or cytoplasm in IHC?

Prioritise interpretable nuclear staining: tissue IHC shows a general nuclear pattern (HPA: tissue IHC), while cellular imaging supports nucleoplasm and nuclear bodies (HPA: subcellular). UniProt also describes nucleolar enrichment, nucleoplasmic foci and localisation with DROSHA in GW bodies (UniProt Q8WYQ5: subcellular location). Because chromogenic sections have limited resolution, score nuclear signal consistently rather than requiring visible nucleolar dots in every positive cell (standard IHC practice). If staining appears predominantly diffuse in cytoplasm, compare serial sections and a no-primary control before calling it DGCR8; the protein has no transmembrane segment (UniProt Q8WYQ5: topology).

Can a negative stain rule out all DGCR8 isoforms?

No: the record lists 3 DGCR8 isoforms, but the supplied evidence does not map this antibody’s epitope across them (UniProt Q8WYQ5: isoforms; datasheet A00475-1). DGCR8 contains a WW domain at residues 301–334 and RNA-binding domains at 511–578 and 620–685 (UniProt Q8WYQ5: domains). Phosphorylated residues are reported, but their effect on recognition by this antibody is unestablished (UniProt Q8WYQ5: modified residues; datasheet A00475-1). Treat an absent stain as an assay result, document retrieval and controls, and seek epitope mapping or independent validation before claiming isoform-specific loss (standard IHC interpretation).

How should I troubleshoot DGCR8 multiplex IF alongside tissue IHC?

On the separate IF/ICC workflow, pair DGCR8 with a marker that identifies the expected cell population; rectal glandular cells provide a relevant comparison for the selected tissue image (datasheet A00475-1; HPA: rectum glandular cells High). Choose spectrally separated fluorophores and assess unstained tissue autofluorescence before assigning faint nuclear signal, especially when comparing channels (standard IF practice). Because DGCR8 is principally nuclear and lacks a transmembrane segment, optimise permeabilisation for antibody access to intracellular nuclear epitopes while preserving nuclear structure (UniProt Q8WYQ5: subcellular location and topology; standard IF practice). Use single-stain and no-primary controls to assess bleed-through and background; do not treat the paraffin IHC caption as IF fixation evidence (standard IF practice; datasheet A00475-1).

What should I check when DAB obscures DGCR8 nuclear staining?

First compare a no-primary section with the test section to distinguish detection-system background from antibody-associated staining (standard IHC practice). The selected image used a biotinylated goat anti-rabbit secondary, a streptavidin–biotin complex and DAB, so include controls suited to that detection sequence (datasheet A00475-1; standard IHC practice). Apply a peroxidase block and assess endogenous biotin where relevant; these are general chromogenic IHC precautions, not demonstrated DGCR8-specific requirements (standard IHC practice). If background persists, check blocking, washing, antibody concentration and DAB development while preserving the documented citrate pH 6 retrieval as the initial reference condition (datasheet A00475-1; standard IHC practice).

How should I score DGCR8 across paraffin tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION

Define the cell population and nuclear scoring rules before comparing sections; HPA reports high staining in rectal glandular cells and a general nuclear pattern (HPA: rectum glandular cells High; HPA: tissue IHC). Report the percentage of evaluable cells with nuclear signal and, when intensity categories are reproducible, an H-score calculated from the percentage in each category (standard IHC quantification). Normalise counts to the number of evaluable cells or sampled tissue area, and report excluded necrotic or damaged regions separately (standard IHC quantification). Keep retrieval, antibody incubation, DAB development and image thresholds matched across samples; document the selected workflow’s 1 μg/mL overnight incubation when using it (datasheet A00475-1; standard IHC practice).

When is an apparent DGCR8-positive section convincing rather than artefactual?

A convincing result shows reproducible nuclear signal in intact cells, consistent with general nuclear tissue expression and supported nucleoplasm or nuclear-body localisation (HPA: tissue IHC; HPA: subcellular). Use the expected cell population as context: HPA reports high rectal glandular staining, whereas adipocytes in adipose tissue were not detected (HPA: rectum glandular cells High; HPA: adipose tissue adipocytes Not detected). Check whether signal concentrates at section edges, in necrosis, or in compartments inconsistent with the nuclear pattern, and compare a no-primary control for endogenous detection activity (standard IHC interpretation). HPA rates the tissue staining Approved with medium consistency against RNA expression, so resolve discordant results with controls before making a biological claim (HPA: tissue IHC reliability).

Boster reagents

Best DGCR8 / Microprocessor complex subunit DGCR8 IHC Antibodies

A00475-1 has paraffin-section IHC images from human rectal cancer and mouse small intestine, plus an IF image from 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 lists IHC and IF/ICC applications, with IHC images from human rectal cancer and mouse small intestine paraffin sections and an IF image from A431 cells (catalog applications and image captions). M00475 is a rabbit monoclonal listed for ICC/IF, with Human, Mouse and Rat reactivity; no IHC application or image is listed (catalog).

Which to pick: Choose A00475-1 for tissue IHC: its own captions document paraffin sections, citrate retrieval and 1 μg/ml primary antibody (A00475-1 IHC image captions); the fixative is unreported. For IF/ICC, A00475-1 has an A431 cell image at 2 μg/ml (A00475-1 IF image caption), while M00475 offers a rabbit monoclonal listed for ICC/IF without an image (M00475 catalog). For cross-species planning, both list Human, Mouse and Rat reactivity, but the supplied IHC images document Human and Mouse only (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.