CREBZF / CREB/ATF bZIP transcription factor · IHC design guide

Design Immunohistochemistry for CREBZF

Plan chromogenic CREBZF IHC on paraffin sections using the catalog antibody’s 1:100–1:300 IHC dilution range (datasheet A07952-2). Expect cytoplasmic staining in many tissues, but interpret it cautiously because tissue IHC reliability is uncertain (HPA tissue IHC).

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

Immunohistochemistry protocol sheet for CREBZF (IHC for CREBZF): expected localisation Cytoplasmic tissue staining; nuclear annotation (HPA tissue IHC; UniProt), antibody A07952-2, validated IHC image, and IHC protocol steps — Printable **CREBZF** IHC protocol sheet — expected localisation Cytoplasmic tissue staining; nuclear annotation (HPA tissue IHC; UniProt), antibody A07952-2, controls and protocol steps. Open the full CREBZF IHC guide →

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CREBZF 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	Cytoplasmic tissue staining; nuclear annotation (HPA tissue IHC; UniProt)
	Staining pattern	Cytoplasmic staining in most tissues (HPA tissue IHC)
	Antigen retrieval	Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A07952-2)
	Positive control ⓘ	Caudate+4 more · see all
	Negative control ⓘ	Adrenal gland+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	Low staining–RNA concordance makes tissue calls uncertain (HPA tissue IHC)
	Regulation	Adult heart, liver and muscle enriched (UniProt)
	Isoform / epitope	0 annotated isoforms; one full-length chain (UniProt)

Section 1

Recommended CREBZF IHC & IF Protocols

The catalog antibody protocol is paired with 4 published CREBZF IHC protocols covering mouse uterus, mouse ovary, and gastric cancer tissue (PMC5915772; PMC6274897; PMC7085260; PMC3878900).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A07952-2)
Fixation	Image fixative and duration unreported (datasheet A07952-2); verify before use.
Sectioning	4–5 µm sections on charged slides (standard)
Deparaffinisation	Xylene, graded ethanol series to water (standard)
Antigen retrieval	Heat retrieval: Tris-EDTA pH 9.0 (datasheet A07952-2); 20 min, 95–100 °C (standard)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% normal serum of the secondary host, 30 min, room temperature (standard)
Primary antibody	Rabbit anti-CREBZF, 1:100-1:300 (datasheet A07952-2)
Primary incubation	Overnight at 4 °C (standard)
Detection	HRP-polymer secondary, DAB chromogen 5–10 min (standard)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	CREBZF-positive staining in neuronal cells of caudate (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control.

💡Decision noteStart with heat-mediated Tris-EDTA, pH 9.0, for the catalog antibody (datasheet A07952-2). The published protocols use their own stated retrieval conditions (PMC5915772; PMC6274897; PMC3878900).

Section 2

What Is the Expected CREBZF Staining Pattern?

For paraffin-section IHC, HPA reports mostly cytoplasmic CREBZF staining, including medium staining in heart cardiomyocytes, esophageal squamous epithelial cells and kidney glomerular cells (HPA: tissue IHC). UniProt places CREBZF in the nucleus, where it can colocalize in PML nuclear bodies, and reports no transmembrane segment (UniProt Q9NS37). Interpret either compartment cautiously: HPA rates its tissue IHC pattern Uncertain because staining and RNA expression have low consistency (HPA: reliability).

What am I looking at on my slide?
Medium cytoplasmic staining in cardiomyocytes, esophageal squamous epithelial cells or glomerular cells.	This matches reported positive tissue and cell combinations (HPA: tissue IHC, Medium). Record the stained cell type and compartment; the cytoplasmic IHC profile remains Uncertain and does not establish exclusive cytoplasmic localization (HPA: reliability; UniProt Q9NS37: nucleus).
Nuclear staining in cells being assessed for CREBZF.	A nuclear signal is biologically plausible because UniProt places CREBZF in the nucleus and describes colocalization in PML nuclear bodies (UniProt Q9NS37). Tissue IHC alone cannot confirm that fine pattern; compare staining with the cytoplasmic HPA tissue profile and controls (HPA: tissue IHC, Uncertain).
Predominantly cell-surface or extracellular staining, with no convincing intracellular signal.	Treat this as a possible artifact: CREBZF has no transmembrane segment or signal peptide and is annotated as nuclear (UniProt Q9NS37). Check section morphology and controls before assigning the signal to CREBZF.
Strong staining in HPA-undetected cells, or broad diffuse color across cells and surrounding tissue.	Unexpected cell staining may reflect cross-reactivity or endogenous detection activity; diffuse color suggests background. HPA reports no detection in adrenal glandular cells and liver cholangiocytes, but its Uncertain IHC reliability makes either finding a comparison point rather than a definitive negative (HPA: tissue IHC, reliability).
No staining in a comparison section expected to show a signal.	Absence in heart cardiomyocytes or esophageal squamous epithelial cells conflicts with HPA's Medium observations (HPA: tissue IHC). First check whether the expected cells are present, then review detection and controls; an HPA-positive tissue is not a guaranteed positive control because the profile is Uncertain (HPA: reliability).

💡Expected CREBZF appearanceA provisional positive is Medium cytoplasmic staining in HPA-reported cells such as cardiomyocytes or esophageal squamous epithelial cells (HPA: tissue IHC, Uncertain); convincing nuclear signal is plausible (UniProt Q9NS37), while dominant cell-surface or extracellular color is suspect (UniProt Q9NS37: no transmembrane segment or signal peptide).

How each factor affects the staining
Compartment evidence	UniProt annotates nuclear CREBZF and PML nuclear-body colocalization (UniProt Q9NS37), whereas tissue IHC is mostly cytoplasmic and Uncertain (HPA: tissue IHC). Report the observed compartment rather than forcing agreement between the sources.
Cell and tissue selection	HPA reports Medium staining in heart cardiomyocytes, esophageal squamous epithelial cells and kidney glomerular cells (HPA: tissue IHC). Its undetected calls apply to specified cells, such as liver cholangiocytes, not every cell in those organs (HPA: tissue IHC).
Molecular interpretation	CREBZF has no transmembrane segment, signal peptide or annotated processing into separate chains (UniProt Q9NS37). Those features support questioning a dominant surface or extracellular pattern; they do not establish how fixation or antigen retrieval affects IHC staining.
IF/ICC Q&A: does it have the same expected compartment as tissue IHC?	HPA approves nucleoplasm and mitochondria as ICC-IF locations and lists images from HaCaT, HeLa and U2OS (HPA: subcellular). Tissue IHC reports mostly cytoplasmic staining with Uncertain reliability (HPA: tissue IHC); assess IF/ICC on its own guide page.

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
No signal in heart cardiomyocytes despite intact tissue morphology.	The result differs from HPA's Medium cardiomyocyte staining, but the tissue IHC profile is Uncertain (HPA: tissue IHC, reliability); the cause cannot be assigned from this comparison alone.	Confirm that cardiomyocytes were evaluated; inspect the assay's positive and detection controls, then document retrieval and antibody conditions used. Treat these as general IHC checks, not evidence of CREBZF-specific fixation sensitivity.
Only nuclei stain, while expected cytoplasmic staining is absent.	Nuclear CREBZF is plausible (UniProt Q9NS37), but the compartment differs from HPA's mostly cytoplasmic tissue IHC profile (HPA: tissue IHC, Uncertain).	Record the compartment and cell type, compare with a second HPA-reported positive tissue, and review control sections before deciding whether the signal is specific.
Cell surfaces or extracellular material stain more strongly than cells.	That distribution is difficult to reconcile with nuclear annotation and the absence of a signal peptide or transmembrane segment (UniProt Q9NS37).	Inspect morphology and no-primary control staining; review blocking, washing and chromogen development as general IHC background checks before interpreting the result.
Unexpected staining appears in adrenal glandular cells or liver cholangiocytes.	HPA reports those cell populations as Not detected, although its IHC reliability is Uncertain (HPA: tissue IHC, reliability). Cross-reactivity or endogenous detection activity is possible.	Check that the stained cells were identified correctly; compare no-primary and other detection controls, and avoid calling the tissue wholly negative or wholly positive.
Brown color spreads across the section without clear cell boundaries.	Diffuse background can obscure the cell and compartment pattern needed to compare with HPA tissue IHC and UniProt localization (HPA: tissue IHC; UniProt Q9NS37).	Review no-primary control, blocking, washing and chromogen development using standard IHC practice; score only clearly localized cellular staining after background is resolved.
Heart and liver give apparently conflicting results.	UniProt reports abundant adult expression in both organs (UniProt Q9NS37), while HPA reports Medium staining in heart cardiomyocytes and no detection in liver cholangiocytes (HPA: tissue IHC). These refer to different measurements and specified cells.	Score each identified cell population separately and report compartment and intensity. Do not infer that all liver cells lack CREBZF or use cholangiocyte staining to invalidate a heart result.

Source: UniProt Q9NS37 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing CREBZF staining

Immunohistochemical analysis of paraffin-embedded human tonsil. 1, Tris-EDTA, pH9.0 was used for antigen retrieval. 2 Antibody was diluted at 1:200 (4° overnight.3, Secondary antibody was diluted at 1:200 (room temperature, 45min).

A07952-2 · IHCImmunohistochemical analysis of paraffin-embedded human tonsil. 1, Tris-EDTA, pH9.0 was used for antigen retrieval. 2 Antibody was diluted a

A07952-2 · IFImmunofluorescence analysis of HepG2 cells, using CREBZF Antibody. The picture on the right is blocked with the synthesized peptide.

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000137504-CREBZF/tissue/cerebral+cortex#imid_13495343

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000137504-CREBZF/tissue/colon#imid_13495253

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000137504-CREBZF/tissue/liver#imid_13495254

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000137504-CREBZF/tissue/kidney#imid_13495271

HPA · ICC-IF HaCaTHuman Protein Atlas immunofluorescence, HaCaT, antibody HPA063652; green = CREBZF, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

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

Sample controls for CREBZF IHC & IF

🧪Start with esophagus: squamous epithelial cells should show CREBZF staining (HPA: Medium in esophageal squamous epithelial cells). Run adrenal gland as the negative tissue (HPA: Not detected in adrenal gland glandular cells); adjacent esophageal stromal cells that remain unstained can help assess background, but their CREBZF-negative status requires separate validation.

Positive control tissue: Caudate (Neuronal cells, HPA Medium)

Negative control tissue: Adrenal gland (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CREBZF in HaCaT, HeLa, U2OS, with annotated localisation: Nucleoplasm (approved), Mitochondria (approved) (HPA subcellular).

Technical controls: Include a no-primary (secondary-only) control, an isotype control matched to the primary antibody’s host species and immunoglobulin class, and CREBZF-knockout material as a biological negative. For chromogenic IHC, quench endogenous peroxidase and check the esophageal section for background signal.

⚠️Feasibility: A target-specific fixation window or fixation effect is unreported in the supplied evidence, and the selected A07952-2 tissue-IHC caption does not state a fixative. The caption reports Tris-EDTA retrieval at pH 9.0 for a paraffin section (selected-SKU caption: Tris-EDTA, pH 9.0); whether retrieval is essential, or whether frozen sections or IF would be easier, is unreported. Assess surface debris and tissue edges for nonspecific chromogen before scoring esophageal squamous cells.

HPA tissue IHC evidence for CREBZF

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low consistency between antibody staining and RNA expression data. 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
Caudate	Neuronal cells	Medium	Protein (IHC)	HPA →
Cerebellum	Purkinje cells	Medium	Protein (IHC)	HPA →
Cerebral cortex	Neuropil	Medium	Protein (IHC)	HPA →
Esophagus	Squamous epithelial cells	Medium	Protein (IHC)	HPA →
Gallbladder	Glandular cells	Medium	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Adrenal gland	Glandular cells	Not detected	Protein (IHC)	HPA →
Bone marrow	Hematopoietic cells	Not detected	Protein (IHC)	HPA →
Breast	Adipocytes	Not detected	Protein (IHC)	HPA →
Cervix	Glandular cells	Not detected	Protein (IHC)	HPA →
Duodenum	Glandular cells	Not detected	Protein (IHC)	HPA →

Section 3

Advanced CREBZF IHC Tips

Troubleshoot CREBZF staining by checking retrieval, compartment, cell type, controls and scoring before interpreting chromogenic signal.

How should I troubleshoot weak CREBZF staining after antigen retrieval?

Start with heat-mediated retrieval in Tris-EDTA at pH 9.0 for the catalog antibody (datasheet A07952-2). The selected paraffin-section image used that buffer and pH with a 1:200 primary dilution, so reproduce those conditions before changing retrieval chemistry (caption A07952-2). If staining remains weak, compare a modest range of heating times on adjacent sections while keeping cooling, antibody incubation and chromogen development consistent (standard IHC practice). Record nuclear and cytoplasmic signal separately, because UniProt places CREBZF in the nucleus while HPA reports predominantly cytoplasmic tissue staining with uncertain reliability (UniProt Q9NS37 subcellular; HPA tissue IHC).

Could fixation explain weak or uneven CREBZF staining?

Target-specific sensitivity to fixation is unknown: the selected image identifies paraffin sections but does not state the fixative (caption A07952-2). Document the specimen’s actual fixation and processing history, then compare sections processed under matched conditions when investigating variable signal (standard IHC practice). Check whether weak staining tracks with section thickness, tissue edges or processing batches before attributing it to CREBZF biology (standard IHC practice). The caption supports Tris-EDTA retrieval at pH 9.0 and a 1:200 primary dilution, but it provides no basis for a CREBZF-specific fixation adjustment (caption A07952-2).

How should I interpret nuclear and cytoplasmic CREBZF staining?

Score nuclear and cytoplasmic staining separately: UniProt places CREBZF in the nucleus and notes colocalisation with CREB3 and HCFC1 in PML nuclear bodies (UniProt Q9NS37 subcellular). HPA instead describes cytoplasmic staining in most tissues, but rates its tissue IHC reliability uncertain because staining and RNA data have low consistency (HPA tissue IHC). HPA cell imaging also reports approved nucleoplasm and mitochondrial locations, which provides context but does not validate a mitochondrial pattern in paraffin sections (HPA subcellular). Examine morphology and compartment-matched controls before calling diffuse cytoplasmic DAB signal specific; document mixed patterns rather than combining them into one intensity score (standard IHC practice).

Could epitope accessibility or protein processing change my CREBZF IHC result?

The record lists one 1–354 chain, no annotated isoforms, no signal peptide and no transmembrane segment for CREBZF (UniProt Q9NS37 processing; topology; isoforms). Its bZIP domain spans residues 204–267, and phosphoserine at residue 50 is an annotated modification (UniProt Q9NS37 domains; modified residues). Those features do not identify the catalog antibody’s epitope or establish that phosphorylation changes its binding, so avoid assigning a staining difference to either mechanism without epitope evidence (UniProt Q9NS37; caption A07952-2). If sections disagree, compare retrieval and staining batches first, then use an independently mapped antibody or an appropriate specificity control to investigate epitope dependence (standard IHC practice).

How can IF help assess a questionable CREBZF IHC localisation?

On the separate IF/ICC guide, multiplex CREBZF with a marker for the cell population being examined, such as cardiomyocytes in heart, where HPA reports medium tissue staining (HPA tissue IHC). Choose spectrally separated fluorophores and inspect unstained tissue for autofluorescence before assigning a weak signal to CREBZF (standard IF practice). CREBZF has no transmembrane segment, but its reported nucleoplasmic and mitochondrial locations mean permeabilisation must allow access to the compartment containing the antibody’s epitope; that epitope is not specified here (UniProt Q9NS37 topology; HPA subcellular; caption A07952-2). Include single-label and secondary-only controls when comparing IF localisation with chromogenic tissue staining (standard IF practice).

What should I check when CREBZF DAB staining looks diffuse?

Run a no-primary control to assess secondary-reagent and detection background, and check the peroxidase block when using an HRP/DAB workflow (standard IHC practice). Compare staining at tissue edges and damaged areas with intact central regions, since uneven reagent exposure and tissue damage can create misleading colour (standard IHC practice). Recheck washes, primary concentration and chromogen development time; the selected image used a 1:200 primary dilution and a 1:200 secondary dilution for 45 minutes at room temperature (caption A07952-2). Treat diffuse cytoplasmic signal cautiously because HPA reports that pattern while rating its tissue IHC evidence uncertain (HPA tissue IHC).

How should I quantify CREBZF across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION

Define the cell population and compartment before scoring, then report the percentage of positive cells and an intensity-based H-score from 0–300 where appropriate (standard IHC practice). Keep nuclear and cytoplasmic scores separate because their interpretation differs for CREBZF (UniProt Q9NS37 subcellular; HPA tissue IHC). For spatial comparisons, count positive cells per mm² of viable tissue and normalise to the number of eligible cells or sampled area, recording exclusions for damaged regions (standard IHC practice). Apply the same threshold, section sampling plan and staining-run controls across groups, especially given HPA’s uncertain tissue IHC reliability (standard IHC practice; HPA tissue IHC).

How do I distinguish a true CREBZF signal from artefact?

Evaluate whether staining occupies a plausible compartment: UniProt reports nuclear CREBZF, whereas HPA reports largely cytoplasmic tissue staining and approved nucleoplasmic and mitochondrial locations in cell imaging (UniProt Q9NS37 subcellular; HPA tissue IHC; HPA subcellular). Check that positive cells match the population being scored; HPA, for example, reports medium staining in heart cardiomyocytes and no detected staining in liver cholangiocytes (HPA tissue IHC). Discount signal confined to section edges, necrotic regions or a positive no-primary control, and investigate residual endogenous peroxidase in an HRP/DAB assay (standard IHC practice). Reproduce the pattern across sections and appropriate controls before making a biological claim, because HPA rates its tissue IHC evidence uncertain (HPA tissue IHC).

Boster reagents

Best CREBZF / CREB/ATF bZIP transcription factor IHC Antibodies

A07952-2 has IHC data from paraffin-embedded human tonsil (IHC image caption) and IF data from HepG2 cells (IF image caption); listed reactivity is human and mouse (catalog reactivity).

Real IHC data Immunohistochemical analysis of paraffin-embedded human tonsil. 1, Tris-EDTA, pH9.0 was used for antigen retrieval. 2 Antibody was diluted at 1:200 (4° overnight.3, Secondary antibody was diluted at 1:200 (room temperature, 45min).

Anti-Zhangfei CREBZF Antibody

Cat # A07952-2

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A07952-2 will render with a paraffin-embedded human tonsil IHC image (IHC image caption). The same SKU lists IF and IHC applications (catalog applications) and has a HepG2 cell IF image (IF image caption).

Which to pick: Choose A07952-2 for paraffin-section IHC because its own image shows paraffin-embedded human tonsil (IHC image caption); the fixative is unreported (IHC image caption). For cell IF/ICC, A07952-2 has an IF image from HepG2 cells (IF image caption), though ICC is not separately listed (catalog applications). For human and mouse work, A07952-2 lists both species (catalog reactivity), while its supplied images show human tonsil and HepG2 cells only (IHC/IF image captions); it is rabbit-hosted with no clone specified (catalog host/clone).

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

References

UniProt Consortium. UniProt entry Q9NS37 (ZHANG_HUMAN, CREB/ATF bZIP transcription factor).
Human Protein Atlas. CREBZF tissue IHC expression (reliability: Uncertain).
Human Protein Atlas. CREBZF subcellular location (ICC-IF): Localized to the nucleoplasm and mitochondria..
Human Protein Atlas. CREBZF antibody validation summary (2 antibodies).
Regulation of Cyclic AMP-Response Element Binding Protein Zhangfei (CREBZF) Expression by Estrogen in Mouse Uterus. Development & reproduction 2018 — PMC5915772.
Activation of CREBZF Increases Cell Apoptosis in Mouse Ovarian Granulosa Cells by Regulating the ERK1/2 and mTOR Signaling Pathways. International journal of molecular sciences 2018 — PMC6274897.
miRNAs as potential biomarkers for the progression of gastric cancer inhibit CREBZF and regulate migration of gastric adenocarcinoma cells. International journal of medical sciences 2020 — PMC7085260.
CREBZF expression and hormonal regulation in the mouse uterus. Reproductive biology and endocrinology : RB&E 2013 — PMC3878900.
PubMed PMID:10871379 — UniProt-cited evidence.
PubMed PMID:16554811 — UniProt-cited evidence.
PubMed PMID:15489334 — UniProt-cited evidence.