ZBTB20 / Zinc finger and BTB domain-containing protein 20 · IHC design guide

Design Immunohistochemistry for ZBTB20

Plan chromogenic ZBTB20 IHC in paraffin sections around its nuclear tissue pattern (HPA tissue IHC). Compare high-staining adipocytes with bone marrow hematopoietic cells where staining was not detected, using the documented catalog antibody workflow (HPA tissue IHC; datasheet A04274-1).

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

Immunohistochemistry protocol sheet for ZBTB20 (IHC for ZBTB20): expected localisation Nuclear staining in most tissues (HPA tissue IHC), antibody A04274-1, validated IHC image, and IHC protocol steps — Printable **ZBTB20** IHC protocol sheet — expected localisation Nuclear staining in most tissues (HPA tissue IHC), antibody A04274-1, controls and protocol steps. Open the full ZBTB20 IHC guide →

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ZBTB20 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	Nuclear staining in most tissues (HPA tissue IHC)
	Staining pattern	Nuclear staining in cells across most tissues (HPA tissue IHC)
	Antigen retrieval	EDTA pH 8.0 HIER, heat-mediated (datasheet A04274-1)
	Positive control ⓘ	Adipose tissue+4 more · see all
	Negative control ⓘ	Bone marrow+4 more · see all

Important caveats

Reasons your staining may differ from the expected pattern.

	Fixation	Keep fixation consistent across samples (standard IHC practice; not target-specific)
	Caveat	Leukocyte expression may not predict marrow IHC signal (UniProt; HPA tissue IHC)
	Regulation	Expression varies by tissue cell type (HPA tissue IHC)
	Isoform / epitope	2 isoforms; epitope coverage is unspecified (UniProt)

Section 1

Recommended ZBTB20 IHC & IF Protocols

The catalog antibody’s IHC-P protocol (datasheet: A04274-1) is paired with a published protocol for HCC and peritumoral liver sections (PMC3145616).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded mouse brain tissue; fixative not specified (datasheet A04274-1)
Fixation	Image fixative and duration unreported (datasheet A04274-1); verify before use.
Sectioning	4–5 µm sections on charged slides (standard)
Deparaffinisation	Xylene, graded ethanol series to water (standard)
Antigen retrieval	Heat retrieval: EDTA pH 8.0 (datasheet A04274-1); 20 min, 95–100 °C (standard)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% goat serum (datasheet A04274-1)
Primary antibody	Rabbit anti-ZBTB20, 1:50 recommended; image 1:100 (datasheet A04274-1)
Primary incubation	Overnight at 4 °C (datasheet A04274-1)
Detection	HRP-conjugated secondary, DAB chromogen (datasheet A04274-1)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	ZBTB20-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in most tissues. No signal in the no-primary control.

💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet: A04274-1); the published protocol used the same pH in a pressure cooker (PMC3145616).

Section 2

What Is the Expected ZBTB20 Staining Pattern?

ZBTB20 is a nuclear, non-membrane protein (UniProt Q9HC78: nucleus; no transmembrane segment). In paraffin-section IHC, expect staining in nuclei of cells reported as positive, including adipocytes, adrenal glandular cells and endometrial stromal cells (HPA tissue IHC: High). HPA describes nuclear expression in most tissues, but its tissue profile is Approved with external verification pending (HPA tissue IHC: reliability and profile).

What am I looking at on my slide?
Strong staining confined mainly to nuclei of adipocytes or adrenal glandular cells.	This fits the reported compartment and cell populations (UniProt Q9HC78: nucleus; HPA tissue IHC: High in adipocytes and adrenal glandular cells). Compare positive nuclei with nearby unstained structures on the same section before scoring intensity (general IHC practice).
Predominantly cytoplasmic or membrane-like staining, with little nuclear signal.	Treat this as a localisation mismatch: ZBTB20 is nuclear and has no transmembrane segment (UniProt Q9HC78: localisation and topology). Review staining specificity and section-wide background before interpreting it as target expression (general IHC practice).
Prominent staining in duodenal glandular cells or bone-marrow hematopoietic cells.	These cell populations were reported as Not detected (HPA tissue IHC: duodenum and bone marrow). Unexpected staining raises concern for cross-reactivity or endogenous chromogenic activity; it does not establish either cause on its own (general IHC practice).
Diffuse colour across nuclei, cytoplasm and tissue spaces.	A section-wide haze makes the expected nuclear pattern hard to judge (UniProt Q9HC78: nucleus; general IHC practice). Check the no-primary control and detection background, then reassess whether any nuclear signal remains distinct from the haze (general IHC practice).
No nuclear signal in a section chosen for a reported High cell population.	An absent signal calls for a technical check, but HPA High is an observed pattern, not a guarantee for every specimen (HPA tissue IHC: High entries; Approved, pending external verification). Check tissue preservation, retrieval and detection controls before calling the specimen negative (general IHC practice).

💡Expected ZBTB20 appearanceCall a result positive when distinct nuclear staining occurs in an HPA-reported High cell population, such as adipocytes; dominant cytoplasmic staining or colour in HPA Not detected cells is suspect (HPA tissue IHC: adipocytes High; duodenal glandular cells Not detected; UniProt Q9HC78: nucleus).

How each factor affects the staining
Cell population used for interpretation	HPA reports High staining in adipocytes, glial cells and several glandular or stromal populations, while selected epithelial and hematopoietic populations are Not detected (HPA tissue IHC). Judge the named cell population, rather than treating an entire tissue as uniformly positive or negative (general IHC practice).
Antibody evidence	The tissue profile is Approved, with external verification pending (HPA tissue IHC: reliability). HPA016815 has an Approved IHC annotation; HPA015551 has no IHC annotation in the supplied record (HPA antibodies). Those statuses should not be treated as equivalent IHC validation.
Isoforms and epitope coverage	UniProt lists 2 ZBTB20 isoforms and a BTB domain at residues 104–167 (UniProt Q9HC78: isoforms and domains). No antibody epitope is supplied, so these facts alone cannot establish which isoform an IHC antibody detects or explain a missing stain.
IF/ICC Q: Where should signal appear?	A: HPA reports enhanced nucleoplasmic and nuclear-body localisation in ICC-IF (HPA subcellular: main location). That observation supports a nuclear expectation, but it is separate from the paraffin-section IHC tissue pattern (HPA tissue IHC: nuclear expression in most tissues).

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
A reported High population has no visible nuclear stain.	The assay may have failed, or the sampled cells may differ from HPA's observed examples (general IHC practice; HPA tissue IHC: High entries, external verification pending).	Check section identity and morphology, then review retrieval, primary-antibody and chromogenic detection controls before interpreting absence (general IHC practice).
Most colour is cytoplasmic or outlines cell borders.	That pattern conflicts with the reported nuclear location and non-membrane topology (UniProt Q9HC78: nucleus; no transmembrane segment).	Compare with a no-primary control, inspect nuclear counterstaining and reassess specificity before scoring the colour as ZBTB20 (general IHC practice).
A reported Not detected population stains strongly.	Cross-reactivity or endogenous detection activity is possible; the HPA observation alone cannot identify the mechanism (HPA tissue IHC: Not detected entries; general IHC practice).	Examine no-primary and detection controls and compare the staining compartment with nuclei in a reported High population (general IHC practice; HPA tissue IHC: High entries).
The whole section has diffuse brown background.	Background from blocking, detection reagents or chromogen can obscure nuclear contrast (general IHC practice).	Review no-primary staining, blocking and wash steps, then judge signal only where nuclei remain clearly distinguishable (general IHC practice; UniProt Q9HC78: nucleus).
Nuclear intensity varies among cells in one tissue.	HPA levels refer to named cell populations and do not specify uniform staining of every cell (HPA tissue IHC: cell-specific levels).	Score the relevant cell type and record the distribution and intensity separately; compare like cells across sections (general IHC practice).
An IF/ICC image looks punctate within nuclei.	Nuclear-body signal can accompany nucleoplasmic signal in ICC-IF (HPA subcellular: enhanced nuclear bodies and nucleoplasm).	Interpret the image against the ICC-IF localisation record; use the separate IF/ICC guide for that application rather than treating puncta as an IHC protocol decision (HPA subcellular: main location; general IHC practice).

Source: UniProt Q9HC78 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing ZBTB20 staining

IHC analysis of ZBTB20 using anti-ZBTB20 antibody (A04274-1). ZBTB20 was detected in a paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:100 rabbit anti-ZBTB20 Antibody (A04274-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

A04274-1 · IHCIHC analysis of ZBTB20 using anti-ZBTB20 antibody (A04274-1). ZBTB20 was detected in a paraffin-embedded section of mouse brain tissue. Heat

IHC analysis of ZBTB20 using anti-ZBTB20 antibody (A04274-1). ZBTB20 was detected in a paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:100 rabbit anti-ZBTB20 Antibody (A04274-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

A04274-1 · IHCIHC analysis of ZBTB20 using anti-ZBTB20 antibody (A04274-1). ZBTB20 was detected in a paraffin-embedded section of rat brain tissue. Heat m

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000181722-ZBTB20/tissue/cerebral+cortex#imid_4307552

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000181722-ZBTB20/tissue/colon#imid_4307470

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000181722-ZBTB20/tissue/liver#imid_4307487

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000181722-ZBTB20/tissue/kidney#imid_4307480

HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA015551; green = ZBTB20, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

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

Sample controls for ZBTB20 IHC & IF

🧪Run cerebral cortex first and look for nuclear staining in glial cells (HPA: High in cerebral cortex glial cells); run bone marrow as the negative tissue, focusing on hematopoietic cells (HPA: Not detected in bone marrow hematopoietic cells). On the cortex slide, use morphologically identified cells with unstained nuclei as internal negatives only after observing that they lack signal; the supplied HPA row assigns a level to glial cells, not to other cortical cell types (HPA: High in cerebral cortex glial cells).

Positive control tissue: Adipose tissue (Adipocytes, HPA High)

Negative control tissue: Bone marrow (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ZBTB20 in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (enhanced), Nuclear bodies (enhanced) (HPA subcellular).

Technical controls: Include a no-primary, secondary-only control; a concentration-matched nonimmune rabbit IgG isotype control (caption: rabbit primary antibody); and ZBTB20-knockout tissue, if available, as a biological negative (standard IHC practice). Quench endogenous peroxidase before DAB detection and inspect cortex for pigment that could be mistaken for staining (standard IHC practice; caption: HRP/DAB detection).

⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A04274-1 paraffin-section caption does not state a fixative (caption: fixative unreported). The mouse-brain IHC caption used heat retrieval in EDTA at pH 8.0, followed by primary antibody at 1:100 overnight at 4°C; treat these as starting conditions for cortex and verify nuclear localisation (caption: retrieval and incubation conditions; UniProt Q9HC78: nucleus). The supplied evidence does not establish whether frozen sections or tissue IF are easier; cortical pigment can complicate chromogenic interpretation, while autofluorescence would need checking in IF (standard IHC/IF practice).

HPA tissue IHC evidence for ZBTB20

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Pending external verification.). 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
Adipose tissue	Adipocytes	High	Protein (IHC)	HPA →
Adrenal gland	Glandular cells	High	Protein (IHC)	HPA →
Caudate	Glial cells	High	Protein (IHC)	HPA →
Cerebellum	Cells in molecular layer	High	Protein (IHC)	HPA →
Cerebral cortex	Glial cells	High	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Bone marrow	Hematopoietic cells	Not detected	Protein (IHC)	HPA →
Cervix	Squamous epithelial cells	Not detected	Protein (IHC)	HPA →
Duodenum	Glandular cells	Not detected	Protein (IHC)	HPA →
Esophagus	Squamous epithelial cells	Not detected	Protein (IHC)	HPA →
Oral mucosa	Squamous epithelial cells	Not detected	Protein (IHC)	HPA →

Section 3

Advanced ZBTB20 IHC Tips

Troubleshoot ZBTB20 staining in paraffin sections by checking retrieval, nuclear localisation, cell identity, and controls before scoring chromogenic signal.

What retrieval should I try first when ZBTB20 nuclear staining is weak?

Start with heat-mediated retrieval in EDTA pH 8.0 for paraffin sections (datasheet A04274-1). The selected mouse brain image used this retrieval, followed by 1:100 primary antibody overnight at 4°C (datasheet A04274-1); compare those conditions with your run before changing the buffer. If staining remains weak, test another retrieval buffer on adjacent sections while holding antibody concentration and detection constant (standard IHC practice). Judge improvement by stronger nuclear signal without increased diffuse background or tissue damage, because ZBTB20 is nuclear (UniProt Q9HC78) and HPA reports nuclear expression in most tissues (HPA tissue IHC).

Could fixation explain weak or uneven ZBTB20 staining?

The selected paraffin-section caption does not state a fixative, so target-specific fixation sensitivity is unknown (datasheet A04274-1). Record the fixative, fixation duration, tissue thickness, and processing history for each block, then compare sections processed together before attributing weak staining to ZBTB20 biology (standard IHC practice). Keep retrieval at EDTA pH 8.0 and the starting primary dilution at 1:100 during that comparison (datasheet A04274-1). If matched sections differ, optimize processing systematically and use nuclear morphology and tissue integrity to assess the result; neither the HPA expression pattern nor ZBTB20's topology establishes a fixation effect (HPA tissue IHC; UniProt Q9HC78).

Where should convincing ZBTB20 staining appear in a tissue section?

Prioritize signal within intact nuclei: UniProt places ZBTB20 in the nucleus, while HPA describes nuclear expression in most tissues (UniProt Q9HC78; HPA tissue IHC). HPA subcellular images further localize it to nucleoplasm and nuclear bodies (HPA subcellular), although puncta may be difficult to resolve with chromogenic tissue IHC (standard IHC practice). Compare DAB deposits with a light nuclear counterstain and inspect individual cells at higher magnification before calling a region positive (standard IHC practice). Strong diffuse cytoplasmic staining without corresponding nuclear signal warrants review of background and detection controls; ZBTB20 has no transmembrane segment (UniProt Q9HC78).

Could isoforms or epitope access change the staining pattern?

ZBTB20 has 2 annotated isoforms and a BTB domain at residues 104–167 (UniProt Q9HC78). Its recorded modified residues include phosphorylation at 211, 353, 357, 690, and 695 (UniProt Q9HC78), but this payload does not map the catalog antibody's epitope or establish which isoforms it detects. If staining differs between specimens, review any available epitope documentation before assigning the difference to splicing or phosphorylation (standard IHC practice). Keep retrieval at EDTA pH 8.0 and compare adjacent sections under matched detection conditions; staining alone cannot identify an isoform or modification state (datasheet A04274-1; standard IHC practice).

How should I check ZBTB20 by IF alongside the IHC result?

For a separate IF experiment, pair ZBTB20 with a marker for the cell population being assessed and a nuclear counterstain, then compare the same cell class with chromogenic IHC (standard IF practice). HPA reports high ZBTB20 signal in cerebral cortex glial cells and caudate glial cells, while its subcellular data place signal in nucleoplasm and nuclear bodies (HPA tissue IHC; HPA subcellular). Choose a fluorophore channel with low tissue autofluorescence and include single-stain controls for multiplex interpretation (standard IF practice). Apply permeabilisation appropriate to an intracellular nuclear epitope, because ZBTB20 is nuclear and has no transmembrane segment (UniProt Q9HC78); IF fixation and permeabilisation conditions are not supplied here.

How can I reduce diffuse or misleading DAB background?

Begin with the selected paraffin-section conditions: 10% goat serum block, 1:100 primary overnight at 4°C, and secondary incubation for 30 minutes at 37°C (datasheet A04274-1). Add a peroxidase block before DAB development and run a no-primary control to assess endogenous enzyme activity and detection background (standard chromogenic IHC practice). If diffuse signal persists, compare a lower primary concentration, shorter development, and more stringent washes on adjacent sections while preserving the retrieval condition (standard IHC practice; datasheet A04274-1: EDTA pH 8.0). Score nuclear signal separately from diffuse cytoplasmic colour, consistent with ZBTB20's nuclear location (UniProt Q9HC78).

What should I measure when comparing ZBTB20 across specimens? ⚠ ANSWER MARKED FOR VERIFICATION

Define the cell population and region before scoring, then report the percentage of positive nuclei and, when intensity matters, a nuclear H-score (standard IHC practice). Normalize positive-cell counts to all evaluable nuclei in that same cell population, or report positive nuclei per mm² with the sampled area stated (standard IHC practice). Use matched counterstaining, imaging, retrieval, and DAB development across specimens so technical variation does not masquerade as expression change (standard IHC practice). Keep adipocytes and cerebral cortex glial cells in separate strata if both are studied, since HPA reports high signal in each distinct cell type (HPA tissue IHC).

How do I distinguish genuine ZBTB20 staining from artefact?

A credible result aligns DAB with intact nuclei and a defined cell population, consistent with UniProt's nuclear assignment and HPA's tissue profile (UniProt Q9HC78; HPA tissue IHC). Compare expected positive areas, such as cerebral cortex glial cells, with reported undetected populations, such as duodenal glandular cells, without treating either HPA observation as an absolute control (HPA tissue IHC: High in cerebral cortex glial cells; Not detected in duodenal glandular cells). Discount staining concentrated at section edges, folds, or necrotic regions, and inspect no-primary controls for endogenous peroxidase colour (standard IHC practice). Predominantly cytoplasmic colour requires technical review before interpretation because ZBTB20 is nuclear (UniProt Q9HC78).

Boster reagents

Best ZBTB20 / Zinc finger and BTB domain-containing protein 20 IHC Antibodies

A04274-1 has IHC images from paraffin sections of mouse and rat brain and human liver; IF/ICC is listed as an application, with no IF image supplied (catalog: IHC image captions; applications; IF image alts).

Real IHC data

Anti-ZBTB20 Antibody

Cat # A04274-1

View product →

A04274-1 is the sole card and lists IHC, IF and ICC for human, mouse and rat samples (catalog: applications; reactivity). Its IHC captions show paraffin sections of mouse and rat brain and human liver; no IF image is supplied (catalog: IHC image captions; IF image alts).

Which to pick: Choose A04274-1 for tissue IHC: its own images show chromogenic staining of paraffin sections after EDTA pH 8.0 retrieval; the fixative is unreported (catalog: A04274-1 IHC image captions). For IF/ICC, A04274-1 lists both applications, but no IF image is supplied (catalog: applications; IF image alts). It is a polyclonal option across human, mouse and rat (catalog: dilution_raw; reactivity).

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

References

UniProt Consortium. UniProt entry Q9HC78 (ZBT20_HUMAN, Zinc finger and BTB domain-containing protein 20).
Human Protein Atlas. ZBTB20 tissue IHC expression (reliability: Approved).
Human Protein Atlas. ZBTB20 subcellular location (ICC-IF): Localized to the nucleoplasm and nuclear bodies..
Human Protein Atlas. ZBTB20 antibody validation summary (2 antibodies).
Zinc finger protein ZBTB20 expression is increased in hepatocellular carcinoma and associated with poor prognosis. BMC cancer 2011 — PMC3145616.
Expression of Transcription Factor ZBTB20 in the Adult Primate Neurogenic Niche under Physiological Conditions or after Ischemia. Genes 2022 — PMC9498320.
Erythropoietin alleviates syndrome-associated intellectual disability and autism-like behavior in Zbtb20-haploinsufficient Primrose syndrome mouse model. JCI insight 2026 — PMC12956014.
ZBTB20 in Nociceptive Neurons of the Trigeminal Ganglia Regulates Pruritus. Frontiers in medicine 2021 — PMC7969640.
PubMed PMID:11352661 — UniProt-cited evidence.
PubMed PMID:11230166 — UniProt-cited evidence.
PubMed PMID:17974005 — UniProt-cited evidence.