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BLZF1 / Golgin-45 · IHC design guide
Design Immunohistochemistry for BLZF1
Plan BLZF1 paraffin IHC using colon glandular cells as a high-staining reference (HPA tissue IHC). Start with the catalog antibody at 1:100–1:300 and include a peptide-absorbed negative control (datasheet A09997).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
BLZF1 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 and cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | Colon glandular cells show nuclear and cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 8.0 HIER, heat-mediated (datasheet A09997) | |
| Positive control | Cerebral cortex+4 more · see all | |
| Negative control | Adipose tissue |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific); optimize empirically. | |
| Caveat | A discrete Golgi pattern may be hard to distinguish in tissue IHC (HPA tissue IHC; UniProt) | |
| Regulation | Low tissue specificity (HPA tissue RNA) | |
| Isoform / epitope | 2 isoforms; epitope impact is undetermined (UniProt) |
Section 1
Recommended BLZF1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A09997) and one published BLZF1 IHC protocol (PMC13106729) provide starting conditions for paraffin sections.
| Sample | Paraffin-embedded Human brain tissue; fixative not specified (datasheet A09997) |
| Fixation | Image fixative and duration unreported (datasheet A09997); 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 8.0 (datasheet A09997); 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-BLZF1, 1:100 - 1:300 (datasheet A09997) |
| 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 | BLZF1-positive staining in neuropil of cerebral cortex (HPA tissue IHC: High). HPA tissue profile: General nuclear and cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated Tris-EDTA retrieval at pH 8.0 for the catalog antibody (datasheet A09997); the published protocol used citrate buffer under high temperature and pressure (PMC13106729).
Section 2
What Is the Expected BLZF1 Staining Pattern?
BLZF1 is mainly Golgi associated, with additional nucleoplasmic localization in ICC-IF (HPA: subcellular). Paraffin tissue IHC shows broader nuclear and cytoplasmic staining (HPA: tissue IHC). Expect staining in selected epithelial, tubular, alveolar, neural and germ cells listed below (HPA: tissue IHC). UniProt annotates a Golgi membrane association without a transmembrane segment (UniProt Q9H2G9 topology). HPA rates the tissue pattern Supported, with medium consistency between staining and RNA data (HPA: tissue IHC).
| Distinct staining in colon or duodenal glandular cells, kidney tubular cells, or esophageal squamous cells. | These are reported High IHC sites (HPA: tissue IHC). Score the named cell population against adjacent tissue and the negative control. A nuclear and cytoplasmic appearance fits the reported paraffin-section profile (HPA: tissue IHC); a resolved Golgi pattern is not required for an IHC-positive call. |
| Staining appears only in an unexpected compartment, with no convincing signal in the expected cells. | Check identity and specificity before interpreting it as BLZF1. Tissue IHC reports general nuclear and cytoplasmic expression (HPA: tissue IHC), while ICC-IF places BLZF1 mainly at the Golgi and additionally in nucleoplasm (HPA: subcellular). The two imaging methods offer different spatial detail. |
| Strong staining is confined to a cell population outside the reported positive populations, or adipocytes stain strongly. | Consider cross-reactivity or endogenous detection activity. Adipocytes are reported Not detected (HPA: adipose tissue IHC), whereas the named positive cells have tissue-specific observations (HPA: tissue IHC). Compare a no-primary control before assigning the unexpected signal to BLZF1 (general IHC practice). |
| Pale, diffuse chromogen covers many cells and the extracellular area, obscuring cell boundaries. | Treat this as background until controls establish a cellular pattern (general IHC practice). The HPA tissue profile describes nuclear and cytoplasmic expression, not an indiscriminate haze (HPA: tissue IHC). Review blocking, washes, detection reagent exposure and chromogen development using the same section conditions. |
| No interpretable staining appears in a section containing a reported High cell population. | A negative run in colon glandular cells or kidney tubular cells is inconclusive without a functioning positive control (HPA: tissue IHC; general IHC practice). Review section quality, the catalog antibody's IHC-P instructions, detection reagents and counterstain before concluding that the specimen lacks BLZF1. |
💡Expected BLZF1 appearanceCall a convincing positive when the appropriate cells show clear nuclear and cytoplasmic chromogen, with strong signal possible in reported High populations such as colon glandular cells (HPA: tissue IHC); uniform haze or strong adipocyte staining warrants investigation (HPA: adipose tissue IHC; general IHC practice).
| Which tissue gives an informative comparison? | Colon and duodenal glandular cells, kidney tubular cells, and testis pachytene spermatocytes are reported High; adrenal glandular cells are Medium; adipocytes are Not detected (HPA: tissue IHC). Compare cell types within a section rather than treating an entire organ as uniformly positive or negative. |
| Does IF/ICC show the same level of detail? | ICC-IF places BLZF1 mainly at the Golgi apparatus and additionally in nucleoplasm (HPA: subcellular). Paraffin tissue IHC is summarized as general nuclear and cytoplasmic staining (HPA: tissue IHC). Use the separate IF/ICC guide for that application; a Golgi-resolved pattern is not a prerequisite for interpreting chromogenic IHC. |
| How strong is the antibody-pattern evidence? | HPA lists IHC as Supported for HPA025703 and HPA027331, and ICC as Supported for HPA067113 (HPA: antibodies). Its overall tissue reliability is Supported with medium staining-to-RNA consistency (HPA: tissue IHC). These ratings support comparison with the observed pattern; they do not establish specificity in every specimen. |
| Can topology or processing predict section behavior? | BLZF1 has no transmembrane segment, signal peptide, propeptide, or annotated glycosylation site; the annotated chain spans residues 1–400 (UniProt Q9H2G9). These facts do not identify the antibody epitope or establish antigen-retrieval needs or fixation sensitivity. Target-specific fixation sensitivity is unreported in the supplied sources. |
| Do variants or modifications change the expected stain? | UniProt lists two isoforms and modified residues including phosphoserines and a phosphothreonine (UniProt Q9H2G9). The supplied sources do not map the catalog antibody epitope to an isoform or modification. Do not attribute an unusual cellular pattern or a negative section to either without separate evidence. |
| Situation | Likely cause | Next action |
|---|---|---|
| Reported High cells are unstained. | The run may have failed, or the chosen section may lack an adequate internal positive population (HPA: tissue IHC; general IHC practice). | Check a section containing a named High population, such as colon glandular cells, and verify the catalog antibody's IHC-P conditions and detection controls (HPA: tissue IHC; general IHC practice). |
| All cell types carry similar weak chromogen. | Nonspecific reagent binding, excessive detection signal or insufficient washing can produce diffuse background (general IHC practice). | Compare the no-primary control; review blocking, washes and development time before scoring cell-specific staining (general IHC practice). |
| Adipocytes stain as strongly as reported positive cells. | Cross-reactivity or endogenous detection activity is possible; adipocytes are reported Not detected (HPA: adipose tissue IHC). | Check the no-primary and detection controls, then reassess whether staining tracks cell boundaries and the expected positive populations (general IHC practice; HPA: tissue IHC). |
| A nuclear or cytoplasmic IHC signal lacks a crisp Golgi shape. | HPA describes general nuclear and cytoplasmic tissue IHC, while its Golgi localization comes from ICC-IF (HPA: tissue IHC; HPA: subcellular). | Judge the paraffin section against the reported IHC cell populations and controls; use the separate IF/ICC guide if resolving the Golgi compartment is the experimental aim. |
| Only the counterstain is visible in both specimen and control. | A missing or inactive primary or detection step is possible (general IHC practice). | Verify reagent order and activity against a known-positive section before interpreting BLZF1 expression in the specimen (general IHC practice; HPA: tissue IHC). |
| The cell pattern differs between runs. | Variation in section selection, staining steps or scoring can change the apparent result (general IHC practice). HPA reports medium staining-to-RNA consistency (HPA: tissue IHC). | Repeat with matched controls and record the named cell type, compartment and intensity in each section; compare the same populations across runs (general IHC practice; HPA: tissue IHC). |
Experiment evidence
IHC & IF images showing BLZF1 staining
A09997 · IHCImmunohistochemical analysis of paraffin-embedded Human brain. Antibody was diluted at 1:100(4° overnight). High-pressure and temperature Tr
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000117475-BLZF1/tissue/cerebral+cortex#imid_6863384
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000117475-BLZF1/tissue/colon#imid_6863310
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000117475-BLZF1/tissue/liver#imid_6863319
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000117475-BLZF1/tissue/kidney#imid_6863312
HPA · ICC-IF U-251MGHuman Protein Atlas immunofluorescence, U-251MG, antibody HPA067113; green = BLZF1, blue = nucleus, red = microtubules.
HPA · ICC-IF U-251MGHuman Protein Atlas immunofluorescence, U-251MG, antibody HPA067113; green = BLZF1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for BLZF1 IHC & IF
🧪Run colon first: glandular cells should stain (HPA: High in colon glandular cells). Run adipose tissue as the negative comparator (HPA: Not detected in adipocytes); on the colon slide, neighboring cells that remain unstained should show only background signal, but their BLZF1-negative status is unconfirmed.
Positive control tissue: Cerebral cortex (Neuropil, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show BLZF1 in U-251MG, U2OS, KOLF2.1J, with annotated localisation: Golgi apparatus (supported) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, a host-species- and antibody-class-matched nonimmune IgG control (monoclonal isotype or polyclonal IgG, as appropriate), and a knockout or immunogen-peptide-block control (A09997 caption: peptide-block control). For colon chromogenic IHC, quench endogenous peroxidase and check endogenous biotin if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: A09997 shows staining of paraffin-embedded brain after high-pressure, high-temperature Tris–EDTA retrieval at pH 8.0, but this does not establish that retrieval is required for BLZF1 (A09997 tissue-IHC caption). The target-specific fixation window and fixation effect are unreported, and the caption does not state a fixative (A09997 tissue-IHC caption). Frozen sections are not established as easier by the supplied evidence; IF/ICC has supported Golgi localization, while colon IHC should be checked for luminal mucus trapping chromogen (HPA: Golgi apparatus, supported; standard IHC practice).
HPA tissue IHC evidence for BLZF1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — 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 |
|---|---|---|---|---|
| Cerebral cortex | Neuropil | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced BLZF1 IHC Tips
Troubleshoot BLZF1 staining in paraffin sections by checking retrieval, tissue preservation, compartment pattern and controls before interpreting chromogenic signal.
Which retrieval conditions should I try first for weak BLZF1 staining?
Use heat-mediated Tris-EDTA pH 8.0 retrieval for paraffin sections (datasheet A09997); the selected human brain image used high-pressure, high-temperature retrieval (caption A09997). Start with the catalog antibody at 1:100 and 4 °C overnight, then change only retrieval duration across matched sections (caption A09997; standard IHC practice). If signal is weak, compare 10 versus 20 minutes of heating with identical cooling and chromogenic detection, checking tissue integrity after each condition (standard IHC practice). Favor crisp perinuclear staining over uniform haze, and compare with the peptide-preabsorbed negative control described for this antibody (HPA subcellular; caption A09997).
How should I investigate fixation as a cause of weak BLZF1 staining?
The selected paraffin-section caption does not report a fixative, so BLZF1-specific fixation sensitivity remains unknown (caption A09997). Compare documented fixation conditions on matched specimens while holding section thickness, retrieval, antibody incubation and detection constant (standard IHC practice). Record fixative type, fixation duration, processing delay and section age; for routinely formalin-fixed material, begin with pH 8.0 retrieval and the reported 1:100 antibody dilution (standard IHC practice; datasheet A09997; caption A09997). Judge any improvement by preserved morphology and localized signal, using the caption’s peptide-preabsorbed section as the available antibody-specific comparator (HPA subcellular; caption A09997).
Should I score nuclear or diffuse cytoplasmic BLZF1 staining as positive?
Prioritize a compact perinuclear pattern consistent with Golgi localization when reviewing chromogenic sections (HPA subcellular; UniProt Q9H2G9 subcellular location). Additional nucleoplasmic signal is supported in subcellular imaging, while nuclear detection in the UniProt record includes heterologous expression (HPA subcellular; UniProt Q9H2G9 subcellular location). HPA tissue IHC reports general nuclear and cytoplasmic expression, with only Supported reliability and medium agreement with RNA data (HPA tissue IHC). Score nuclear and broad cytoplasmic staining separately from the perinuclear pattern, and examine each against morphology, local background and a matched negative control (standard IHC practice).
Could isoforms or epitope accessibility explain inconsistent BLZF1 staining?
BLZF1 has 2 annotated isoforms, but the supplied antibody evidence does not locate its epitope or establish isoform coverage (UniProt Q9H2G9 isoforms; caption A09997). Its sequence has no annotated transmembrane segment, and recorded phosphorylation sites include residues 15, 49, 348 and 353 (UniProt Q9H2G9 topology; modified residues). Those facts alone cannot establish whether fixation, retrieval or phosphorylation changes this antibody’s binding in sections (UniProt Q9H2G9; standard IHC interpretation). Compare matched sections under the reported pH 8.0 retrieval conditions and use peptide preabsorption to assess the observed staining pattern (datasheet A09997; caption A09997).
How can IF help resolve an ambiguous BLZF1 IHC pattern?
Use IF as a separate localization check, pairing BLZF1 with an established marker for the expected cell type in the tissue being examined (standard IF practice). For example, HPA reports high staining in kidney tubular cells, so a tubular cell marker can clarify which cells carry the signal (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and inspect unstained tissue for autofluorescence before interpreting apparent overlap (standard IF practice). Because BLZF1 has no annotated transmembrane segment but the antibody epitope’s membrane-facing side is unspecified, compare gentle permeabilization conditions and judge whether signal resolves to Golgi structures (UniProt Q9H2G9 topology; HPA subcellular; standard IF practice).
How do I reduce diffuse chromogenic background without losing BLZF1 signal?
First compare the stained section with a no-primary control and the peptide-preabsorbed control described for A09997 to separate detection background from antibody-dependent staining (caption A09997; standard IHC practice). Block endogenous peroxidase before chromogenic detection, and keep DAB development time identical across comparison sections (standard IHC practice). If haze persists, titrate the catalog antibody around its reported 1:100, 4 °C overnight condition while maintaining the stated pH 8.0 retrieval (caption A09997; datasheet A09997; standard IHC practice). Evaluate background in tissue regions alongside the expected compact Golgi-region pattern, since HPA places BLZF1 mainly at the Golgi apparatus (HPA subcellular; standard IHC practice).
What is a defensible way to quantify BLZF1 IHC staining? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored compartment before analysis, keeping compact perinuclear signal separate from nuclear and diffuse cytoplasmic staining (HPA subcellular; HPA tissue IHC; standard IHC practice). Report the percentage of positive viable cells and an H-score using intensity grades 0–3; if measuring focal structures, report positive area or object density per mm² (standard IHC practice). Normalize counts to the number of eligible cells or viable tissue area within the same annotated region, and apply one staining threshold across the comparison set (standard IHC practice). Include matched negative controls and stratify results by cell type, since HPA tissue staining differs across annotated populations (caption A09997; HPA tissue IHC).
How can I distinguish true BLZF1 staining from artefact?
A credible result follows tissue morphology and shows compartmental enrichment consistent with the mainly Golgi localization, while allowing separately assessed nucleoplasmic signal (HPA subcellular; standard IHC practice). Check the expected cell population: HPA reports high staining in cerebral cortex neuropil and kidney tubular cells, but no detected staining in adipocytes (HPA tissue IHC). Treat staining confined to section edges, folds or necrotic regions as suspect, and check persistent DAB signal in a no-primary section for endogenous enzyme activity (standard IHC practice). Compare any proposed positive pattern with the antibody’s peptide-preabsorbed control before assigning specificity (caption A09997; standard IHC practice).
Boster reagents
Best BLZF1 / Golgin-45 IHC Antibodies
A09997 has an IHC image of a human brain paraffin section (A09997 IHC caption). The catalog lists human and mouse IHC/IF reactivity, plus rat reactivity for A09997-2 (catalog applications/reactivity).
Real IHC data
Anti-BLZF1/Golgin 45 Antibody
Cat # A09997
The rendered SKU, A09997, has an IHC image from a human brain paraffin section (A09997 IHC caption). Its catalog lists IHC, IF and ICC applications for human and mouse samples; no IF image is supplied (A09997 catalog applications/reactivity; catalog image records).
Which to pick: For tissue IHC, start with A09997: its own image documents a human brain paraffin section at 1:100 with Tris-EDTA pH 8.0 retrieval (A09997 IHC caption). For IF/ICC, A09997 lists both applications for human and mouse, while A09997-2 lists IF but not ICC (catalog applications/reactivity). For rat samples, A09997-2 lists rat reactivity and IHC/IF applications (A09997-2 catalog applications/reactivity); the fixative in A09997’s paraffin-section image is unreported (A09997 IHC caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9H2G9 (GO45_HUMAN, Golgin-45).
- Human Protein Atlas. BLZF1 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. BLZF1 subcellular location (ICC-IF): Mainly localized to the Golgi apparatus. In addition localized to the nucleoplasm..
- Human Protein Atlas. BLZF1 antibody validation summary (3 antibodies).
- The role of BLZF1 in lung adenocarcinoma and its value as a diagnostic and prognostic biomarker. Scientific reports 2026 — PMC13106729.
- PubMed PMID:9129147 — UniProt-cited evidence.
- PubMed PMID:11056056 — UniProt-cited evidence.
- PubMed PMID:11739401 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for BLZF1 (Golgin-45). Expected localisation Nuclear and cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 8.0 HIER, heat-mediated (datasheet A09997). Validated IHC antibody: A09997 at 1:100 - 1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Cerebral cortex (Neuropil); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded Human brain tissue; fixative not specified (datasheet A09997); 2. Fixation: Image fixative and duration unreported (datasheet A09997); 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 8.0 (datasheet A09997); 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-BLZF1, 1:100 - 1:300 (datasheet A09997); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); optimize empirically.; Caveat: A discrete Golgi pattern may be hard to distinguish in tissue IHC (HPA tissue IHC; UniProt); Isoform / epitope: 2 isoforms; epitope impact is undetermined (UniProt); Regulation: Low tissue specificity (HPA tissue RNA). Evidence: UniProt Q9H2G9, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the Golgi apparatus. In addition localized to…, Datasheet A09997, PMC13106729. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/BLZF1/immunohistochemistry-ihc-guide.