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CREBZF / CREB/ATF bZIP transcription factor · IHC design guide
Design Immunohistochemistry for CREBZF
Plan chromogenic CREBZF IHC in paraffin sections with the catalog antibody at 1:100–1:300 (datasheet A07952-2). Compare cytoplasmic tissue staining (HPA tissue IHC) with its nuclear molecular annotation (UniProt), and interpret the pattern 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
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 in tissue (HPA tissue IHC); nuclear location (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 | Tissue staining has low concordance with RNA expression (HPA tissue IHC) | |
| Regulation | Adult tissue abundance varies (UniProt) | |
| Isoform / epitope | No annotated isoforms; one 1–354 chain (UniProt) |
Section 1
Recommended CREBZF IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with four published CREBZF IHC methods (PMC5915772; PMC6274897; PMC7085260; PMC3878900).
| 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 Tris-EDTA pH 9.0 heat retrieval for the catalog antibody (datasheet A07952-2); published methods used different retrieval conditions (PMC5915772; PMC6274897; PMC3878900).
Section 2
What Is the Expected CREBZF Staining Pattern?
CREBZF is a nuclear transcription factor with no transmembrane segment (UniProt Q9NS37). In tissue IHC, HPA instead reports mainly cytoplasmic staining, including medium signal in cardiomyocytes and kidney glomerular cells (HPA tissue IHC). Treat that pattern as provisional: HPA rates its tissue IHC reliability Uncertain because antibody staining and RNA expression show low consistency (HPA tissue IHC).
| Medium cytoplasmic staining in cardiomyocytes or kidney glomerular cells. | This matches the reported tissue IHC pattern (HPA tissue IHC: Medium in both). Record the stained cell type and compartment; the pattern is provisional because tissue IHC reliability is Uncertain (HPA tissue IHC). |
| Nuclear signal accompanies cytoplasmic staining in cells of interest. | Nuclear localisation is biologically plausible (UniProt Q9NS37: nucleus), but HPA tissue IHC describes chiefly cytoplasmic staining (HPA tissue IHC). Score the compartments separately and seek independent confirmation before treating either as definitive. |
| Staining is confined to extracellular material or appears as granular deposit outside cells. | That distribution does not match the reported cellular IHC pattern (HPA tissue IHC) or nuclear localisation (UniProt Q9NS37). Check the no-primary control and chromogen distribution; deposition or background is a more useful first interpretation than target localisation (general IHC practice). |
| Strong signal appears in cholangiocytes or adrenal glandular cells. | HPA reports these specific cell populations as Not detected (HPA tissue IHC). Review cell identification and no-primary controls; cross-reactivity or endogenous detection activity is possible (general IHC practice), although HPA's Uncertain rating prevents a definitive false-positive call (HPA tissue IHC). |
| No staining appears in cardiomyocytes or kidney glomerular cells. | These are reported Medium examples (HPA tissue IHC), so inspect positive-control performance, retrieval and detection conditions (general IHC practice). An absent signal alone cannot establish absent CREBZF, given the Uncertain tissue IHC reliability (HPA tissue IHC). |
💡Expected CREBZF appearanceA provisional IHC positive is cellular, predominantly cytoplasmic medium staining in cardiomyocytes or kidney glomerular cells (HPA tissue IHC); extracellular deposits or signal in HPA Not detected cell populations warrant control-based review (HPA tissue IHC; general IHC practice).
| How should the compartment mismatch affect scoring? | UniProt places CREBZF in the nucleus, including PML nuclear bodies (UniProt Q9NS37); HPA tissue IHC describes cytoplasmic expression in most tissues (HPA tissue IHC). Record nuclear and cytoplasmic scores separately, without assuming the IHC pattern resolves the disagreement. |
| Which tissue examples are useful for comparison? | Heart cardiomyocytes and kidney glomerular cells are Medium examples; adrenal glandular cells and liver cholangiocytes are Not detected examples (HPA tissue IHC). Compare the named cell populations, rather than treating an entire organ as uniformly positive or negative. |
| How strong is the antibody evidence? | The tissue IHC profile is Uncertain, with low consistency between staining and RNA expression (HPA tissue IHC). HPA021526 has Uncertain IHC status; HPA063652 has Supported ICC status, which does not validate the tissue IHC pattern (HPA antibodies). |
| What does IF/ICC show? | HPA reports approved nucleoplasm and mitochondria localisation in ICC-IF, with images from HaCaT, HeLa and U2OS (HPA subcellular). This is a separate assay and does not turn mitochondrial staining into an established paraffin-section IHC expectation (HPA subcellular; HPA tissue IHC). |
| Can processing or topology explain membrane staining? | CREBZF has no transmembrane segment, signal peptide, propeptide or reported isoforms, and its annotated chain spans residues 1–354 (UniProt Q9NS37). A membrane-only pattern has no support from those annotations; check controls and antibody specificity (general IHC practice). |
| What is known about fixation sensitivity? | No target-specific fixation effect is reported in the supplied UniProt or HPA evidence. If signal is weak, review the laboratory's general retrieval and detection controls (general IHC practice); do not infer that fixation selectively altered CREBZF staining. |
| Situation | Likely cause | Next action |
|---|---|---|
| Signal covers nuclei, cytoplasm and empty slide areas uniformly. | Nonspecific background, residual reagent or excessive detection can produce widespread colour (general IHC practice). | Compare with the no-primary control; review blocking, washing and detection conditions (general IHC practice). Do not score diffuse slide-wide colour as the HPA cellular pattern (HPA tissue IHC). |
| A tissue expected to be positive has no visible signal. | HPA reports Medium cardiomyocyte and glomerular-cell staining, but rates tissue IHC Uncertain (HPA tissue IHC); technical failure and biological variability remain possible. | Check a same-run positive control, section quality, retrieval and detection steps (general IHC practice). Document the absent signal without converting HPA's example into a guaranteed positive. |
| Only nuclear staining is present in an IHC section. | Nuclear localisation fits UniProt, while HPA tissue IHC chiefly reports cytoplasmic staining (UniProt Q9NS37; HPA tissue IHC). | Score nuclear staining separately; compare cell distribution with controls and, if needed, an independent antibody (general IHC practice). Report the source disagreement rather than silently combining compartments. |
| Signal appears in an HPA Not detected cell population. | Unexpected staining may reflect cell misidentification, cross-reactivity or endogenous detection activity (general IHC practice); the HPA IHC assessment is Uncertain (HPA tissue IHC). | Verify the cell population and inspect no-primary and detection controls (general IHC practice). Treat HPA's Not detected label as a comparison point, not proof that every signal is artefactual. |
| Brown precipitate seems to outline vessels or sit outside cells. | Extracellular deposits are inconsistent with the cellular IHC profile (HPA tissue IHC) and nuclear annotation (UniProt Q9NS37); chromogen artefact is possible (general IHC practice). | Inspect an unstained or no-primary section and check wash and chromogen handling (general IHC practice). Exclude deposits from cellular scoring. |
| ICC-IF shows mitochondrial signal, but IHC scoring expects cytoplasm. | Approved ICC-IF localisation includes mitochondria and nucleoplasm (HPA subcellular); tissue IHC describes broad cytoplasmic expression with Uncertain reliability (HPA tissue IHC). | Keep assay-specific annotations separate. Record the observed IHC compartment and cell type, and use independent validation if a precise mitochondrial claim is needed (general IHC practice). |
Experiment evidence
IHC & IF images showing CREBZF staining
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.
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
🧪Run caudate first and expect staining in neuronal cells (HPA: Medium in caudate neuronal cells). Use adrenal gland glandular cells as the negative tissue (HPA: Not detected); on the caudate slide, treat cells without specific staining above background as internal negative cells, without assuming a particular cell type is negative.
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 and an isotype-matched control appropriate to the primary antibody’s host species and clonality (standard IHC practice). Use a CREBZF knockout specimen or a validated immunizing-peptide block as a biological specificity control, and quench endogenous peroxidase for chromogenic detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the fixative in the selected A07952-2 paraffin-section caption is unreported (A07952-2 tissue-IHC caption). That caption reports Tris-EDTA retrieval at pH 9.0 and primary antibody dilution of 1:200; it does not establish how dependent staining is on retrieval (A07952-2 tissue-IHC caption). ICC-IF images exist in HaCaT, HeLa and U2OS, but the supplied evidence does not establish that frozen sections or IF are easier (HPA subcellular); neuronal lipofuscin can produce autofluorescence in brain tissue (standard IF practice).
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
Use compartment-aware controls when evaluating CREBZF in chromogenic paraffin-section IHC; reported nuclear and tissue-staining patterns require careful interpretation (UniProt Q9NS37; HPA tissue IHC).
Which retrieval conditions should I use when CREBZF staining is weak in paraffin sections?
Use heat-mediated Tris-EDTA at pH 9.0 for CREBZF retrieval in paraffin sections (datasheet A07952-2). The selected human tonsil caption pairs that retrieval with primary antibody at 1:200 overnight at 4°C (caption A07952-2). If staining is weak, compare a small retrieval-time series while holding antibody dilution and detection constant, and check tissue preservation alongside signal (general IHC practice). Include a no-primary control and a consistently processed comparison section so additional heating is not mistaken for improved specificity (general IHC practice). Score nuclear and cytoplasmic staining separately because the reported locations differ across sources (UniProt Q9NS37; HPA tissue IHC).
How should I assess fixation-related loss of CREBZF staining?
Target-specific sensitivity to fixation is unknown: the selected caption identifies paraffin-embedded tonsil but does not state its fixative (caption A07952-2). Record the fixative, fixation duration, tissue thickness, and processing history for each specimen before comparing staining intensity (general IHC practice). If staining varies across a batch, compare similarly processed sections using the specified Tris-EDTA retrieval at pH 9.0 and the same detection settings (datasheet A07952-2; general IHC practice). Inspect morphology and staining gradients before changing retrieval conditions, since uneven processing can distort spatial comparisons (general IHC practice). Do not assign a CREBZF-specific fixation effect without directly matched specimens and controls (general IHC practice).
Should CREBZF staining be nuclear, cytoplasmic, or both?
Evaluate nuclear staining because CREBZF is annotated as nuclear and can colocalize with CREB3 and HCFC1 in PML nuclear bodies (UniProt Q9NS37). Also document cytoplasmic staining separately: tissue IHC reports cytoplasmic expression in most tissues, while subcellular imaging reports nucleoplasm and mitochondria (HPA tissue IHC; HPA subcellular). These observations do not establish which compartment a given chromogenic signal represents, especially because tissue IHC reliability is uncertain (HPA tissue IHC). Compare compartment patterns with matched controls and tissue morphology, and avoid treating isolated diffuse cytoplasmic color as proof of CREBZF (general IHC practice). Report cell type and compartment together for every scored region (general IHC practice).
Could the antibody epitope explain a missing nuclear signal?
The supplied record lists a single 1–354 chain, no isoforms, a bZIP domain at 204–267, and phosphoserine at 50 (UniProt Q9NS37). The antibody epitope is not supplied, so those features cannot establish whether fixation or retrieval masks this antibody's binding site (caption A07952-2; general IHC practice). First repeat the documented Tris-EDTA retrieval at pH 9.0 and check that the primary incubation matches 1:200 overnight at 4°C (datasheet A07952-2; caption A07952-2). If signal remains absent, compare an independently characterized antibody or a loss-of-target control, where available, before assigning the result to an epitope effect (general IHC practice).
How can IF help resolve an ambiguous CREBZF IHC pattern?
Use IF as a follow-up comparison to chromogenic IHC, keeping its controls and interpretation separate (general IHC/IF practice). In heart muscle, multiplex CREBZF with a cardiomyocyte identity marker because cardiomyocytes show medium tissue-IHC staining; interpret that pairing cautiously because the tissue profile has uncertain reliability (HPA tissue IHC). Choose a fluorophore channel after measuring tissue autofluorescence and include single-label controls to check spectral spillover (general IF practice). Permeabilise for intracellular access when assessing nuclear or mitochondrial signal; CREBZF has no transmembrane segment, and both compartments are reported (UniProt Q9NS37; HPA subcellular). Compare those compartments with nuclear counterstaining rather than equating every fluorescent punctum with a PML nuclear body (UniProt Q9NS37; general IF practice).
How do I distinguish CREBZF staining from chromogenic background?
Begin with the documented 1:200 primary dilution and the matched 1:200 secondary incubation for 45 minutes at room temperature, then change one condition at a time (caption A07952-2; general IHC practice). Include a no-primary section to reveal secondary-reagent or detection background, and block endogenous peroxidase before DAB development (general chromogenic IHC practice). Inspect slide edges, damaged regions, and extracellular deposits for staining that does not follow intact cell boundaries (general IHC practice). Compare background with nuclear and cytoplasmic compartments separately because reported CREBZF locations differ (UniProt Q9NS37; HPA tissue IHC). Keep development and counterstaining consistent across comparison sections (general IHC practice).
What should I quantify when CREBZF stains more than one compartment? ⚠ ANSWER MARKED FOR VERIFICATION
Define cell type and compartment before scoring, since nuclear annotation and cytoplasmic tissue-IHC staining do not describe the same readout (UniProt Q9NS37; HPA tissue IHC). For each compartment, report the percentage of positive viable cells and, if intensity is reproducible, an H-score from 0–300 (general IHC practice). For sparse positive cells, report positive-cell density per mm² of evaluable tissue and the area examined (general IHC practice). Normalise cell percentages to the relevant viable cell population, excluding necrosis and tissue-free space from denominators (general IHC practice). Apply identical thresholds across sections and disclose the tissue-IHC profile's uncertain reliability (general IHC practice; HPA tissue IHC).
When is an apparent CREBZF-positive cell likely to be an artefact?
Treat a reproducible intracellular pattern in intact cells as a candidate signal, then compare its compartment with nuclear CREBZF annotation and reported cytoplasmic tissue staining (UniProt Q9NS37; HPA tissue IHC). Check cell identity: heart-muscle cardiomyocytes have medium reported staining, whereas liver cholangiocytes are listed as not detected, although the tissue profile is uncertain (HPA tissue IHC). Suspect artefact when color concentrates at section edges, necrotic areas, or damaged cells, or appears in a no-primary control (general IHC practice). Endogenous enzyme activity can also produce chromogenic signal unless appropriately blocked (general chromogenic IHC practice). Resolve discordant patterns with an independent antibody or loss-of-target control before calling them CREBZF-specific (general IHC practice).
Boster reagents
Best CREBZF / CREB/ATF bZIP transcription factor IHC Antibodies
A07952-2 has IHC data from paraffin-embedded human tonsil and IF data from HepG2 cells (catalog image captions). Listed reactivity covers Human and Mouse (catalog reactivity).
Real IHC data
Anti-Zhangfei CREBZF Antibody
Cat # A07952-2
A07952-2 will render with an IHC image of paraffin-embedded human tonsil (catalog IHC image caption). Its IF image shows HepG2 cells with peptide blocking; IHC and IF are listed applications (catalog IF image caption; catalog applications).
Which to pick: For tissue IHC, choose A07952-2 for paraffin sections: its image caption reports Tris-EDTA pH 9 retrieval and a 1:200 primary dilution; the fixative is unreported (catalog IHC image caption). For IF, A07952-2 has a HepG2 image; ICC validation is unreported, and the antibody is rabbit-hosted with no clone specified (catalog IF image caption; catalog host/clone). For cross-species work, A07952-2 lists Human and Mouse reactivity, while its supplied IHC image documents human tissue only (catalog reactivity; catalog IHC image caption).
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.
Immunohistochemistry protocol sheet for CREBZF (CREB/ATF bZIP transcription factor). Expected localisation Cytoplasmic in tissue (HPA tissue IHC); nuclear location (UniProt). Antigen retrieval: Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A07952-2). Validated IHC antibody: A07952-2 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Caudate (Neuronal cells); negative Adrenal gland. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A07952-2); 2. Fixation: Image fixative and duration unreported (datasheet A07952-2); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: Tris-EDTA pH 9.0 (datasheet A07952-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-CREBZF, 1:100-1:300 (datasheet A07952-2); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Tissue staining has low concordance with RNA expression (HPA tissue IHC); Isoform / epitope: No annotated isoforms; one 1–354 chain (UniProt); Regulation: Adult tissue abundance varies (UniProt). Evidence: UniProt Q9NS37, HPA tissue IHC · Uncertain, HPA ICC-IF · nucleoplasm and mitochondria., Datasheet A07952-2, PMC5915772, PMC6274897. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CREBZF/immunohistochemistry-ihc-guide.