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BLZF1 / Golgin-45 · IHC design guide
Design Immunohistochemistry for BLZF1
Plan BLZF1 chromogenic IHC in paraffin sections using high-staining colon glandular cells and adipocytes with no detected staining as references (HPA tissue IHC). The guide covers antibody titration, fixation consistency, and scoring the observed nuclear and cytoplasmic pattern (HPA tissue IHC).
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/cytoplasmic tissue staining (HPA tissue IHC); Golgi membrane annotated (UniProt) | |
| Staining pattern | Colon glandular cells: nuclear/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) | |
| Caveat | Tissue staining differs from the annotated Golgi location (HPA tissue IHC) (UniProt) | |
| Regulation | Low tissue RNA specificity (HPA tissue RNA) | |
| Isoform / epitope | 2 isoforms; epitope differences unspecified (UniProt) |
Section 1
Recommended BLZF1 IHC & IF Protocols
The catalog antibody’s IHC protocol uses Tris-EDTA retrieval (datasheet A09997). The published lung adenocarcinoma protocol below uses citrate retrieval (PMC13106729).
| 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 at pH 8.0 for the catalog antibody (datasheet A09997); citrate retrieval is the published alternative (PMC13106729).
Section 2
What Is the Expected BLZF1 Staining Pattern?
BLZF1 is mainly Golgi localized in ICC-IF, with additional nucleoplasmic signal (HPA: supported subcellular locations). Tissue IHC shows general nuclear and cytoplasmic staining, with high signal in several epithelial and other cell types (HPA: tissue IHC). The tissue profile is supported with medium consistency between staining and RNA data (HPA: Supported). BLZF1 has no transmembrane segment (UniProt Q9H2G9 topology).
| Nuclear and cytoplasmic staining in colon or duodenal glandular cells, with readily visible signal. | This fits the reported tissue pattern and high staining in those cells (HPA: general nuclear and cytoplasmic expression; High in colon and duodenal glandular cells). IHC alone does not resolve a Golgi pattern. |
| Staining is confined to extracellular spaces or gland lumens, with little cell-associated signal. | That distribution does not match the reported intracellular locations (HPA: tissue IHC; HPA: supported Golgi and nucleoplasm in ICC-IF). Check whether deposits or detection background are being read as positive cells. |
| Adipocytes stain strongly while adjacent expected-positive cells stain weakly or not at all. | Adipocytes were reported as not detected, so this pattern raises concern for cross-reactivity or endogenous detection activity (HPA: Not detected in adipocytes). It does not establish that every adipocyte must be negative. |
| A broad haze covers cells and tissue spaces without clear cellular boundaries. | Diffuse background makes compartment and cell-type scoring unreliable. The reported tissue pattern is cellular (HPA: general nuclear and cytoplasmic expression); review the negative control and detection background before interpreting it. |
| No staining appears in a section containing a reported high-staining cell population. | An absent positive control limits interpretation of the test section (HPA: High in kidney tubular cells, among other listed populations). Check tissue identity, section quality, antibody application and detection before calling BLZF1 absent. |
💡Expected BLZF1 appearanceCall a convincing IHC positive when the expected cells show discernible nuclear and cytoplasmic staining, especially in a listed high-staining population (HPA: tissue IHC); treat isolated extracellular deposits or uniform haze as suspect, since supported ICC-IF places BLZF1 mainly at the Golgi with additional nucleoplasmic signal (HPA: subcellular).
| Tissue and cell selection | Use listed high-staining populations, such as kidney tubular cells or colon glandular cells, to judge an IHC run (HPA: High). Adipocytes are a useful contrasting population, but their reported result is not a universal negative-control guarantee (HPA: Not detected). |
| Antibody evidence | Two listed antibodies have supported IHC status, HPA025703 and HPA027331 (HPA: antibody validation). HPA067113 has supported ICC status and no stated IHC status (HPA: antibody validation); do not transfer that ICC assessment to paraffin IHC. |
| Topology and molecular forms | BLZF1 has no annotated transmembrane segment, no signal peptide or propeptide, and a chain spanning residues 1–400 (UniProt Q9H2G9 topology and processing). These annotations do not establish retrieval conditions or predict staining intensity. |
| Isoforms and modified residues | UniProt lists 2 isoforms and four example phosphorylation sites (UniProt Q9H2G9 isoforms and modified residues). Without an antibody epitope map, these facts cannot predict which forms stain or whether retrieval changes recognition. |
| IF/ICC Q: Should punctate Golgi staining be expected in paraffin IHC? | A: Golgi localization is supported in ICC-IF, alongside nucleoplasmic signal (HPA: supported subcellular locations). Tissue IHC is summarized as general nuclear and cytoplasmic expression (HPA: tissue IHC); do not require resolvable Golgi puncta for an IHC positive. |
| Situation | Likely cause | Next action |
|---|---|---|
| A listed high-staining population is entirely negative. | A failed control, tissue mismatch or an unsuccessful staining step is possible; HPA reports high staining in several specific cell types (HPA: tissue IHC). | Confirm the expected cell population on the section, then review the primary antibody, detection reagents and run controls. Assess retrieval against the antibody's IHC instructions as a general workflow check. |
| The slide shows widespread, low-contrast brown haze. | Nonspecific background or detection activity can obscure cellular staining; the reported IHC distribution is nuclear and cytoplasmic (HPA: tissue IHC). | Inspect a section processed without primary antibody and review blocking, washes and chromogen development. Score cells only when their signal can be separated from background. |
| Only gland lumens or extracellular material are colored. | The distribution conflicts with the reported intracellular pattern (HPA: tissue IHC; HPA: supported Golgi and nucleoplasm in ICC-IF). Deposited chromogen or nonspecific material is possible. | Compare cell boundaries and counterstain with the colored material. Review the negative control before attributing acellular color to BLZF1. |
| Adipocytes appear strongly positive. | This differs from their reported not-detected result (HPA: adipocytes). Cross-reactivity or endogenous detection activity should be considered, especially if expected-positive cells fail. | Check the no-primary control for endogenous detection activity, then compare staining in a listed high-staining population (HPA: tissue IHC). Avoid calling the adipocyte signal specific from this section alone. |
| Golgi puncta are not distinguishable despite cellular IHC staining. | Golgi localization comes from ICC-IF, while the tissue IHC summary is nuclear and cytoplasmic (HPA: subcellular; HPA: tissue IHC). | Score the paraffin IHC against its reported cellular pattern and controls. Reserve a claim of resolved Golgi localization for imaging that actually demonstrates that compartment. |
| Nuclear staining seems stronger than cytoplasmic staining across a sample. | Nuclear staining is within the reported tissue profile and nucleoplasmic localization is supported in ICC-IF (HPA: tissue IHC; HPA: subcellular). Relative intensities are not specified. | Record each compartment separately and compare with appropriate controls. Do not reject the run solely because the nuclear-to-cytoplasmic ratio differs from an assumed value. |
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 and expect staining in glandular cells (HPA: High in colon glandular cells). Use adipose tissue as the negative tissue (HPA: Not detected in adipocytes); within the colon slide, internal negative cells should show counterstain without specific chromogen, but identify them empirically because the HPA row does not establish which nonglandular cells are BLZF1-negative.
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 and a host-species-matched isotype IgG control for a monoclonal antibody, or preimmune IgG for a polyclonal antibody (standard IHC practice). Add a BLZF1 knockout control or peptide-blocked antibody control (A09997 tissue-IHC caption: immunogen-peptide block); for chromogenic colon IHC, quench endogenous peroxidase and inspect inflammatory cells for residual background (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the fixative in the A09997 paraffin-section caption is unreported (A09997 tissue-IHC caption: fixative not stated). That caption uses high-pressure, heated Tris-EDTA at pH 8.0 for retrieval and antibody at 1:100 overnight at 4 °C; these are starting conditions, while a BLZF1-specific retrieval requirement remains unreported (A09997 tissue-IHC caption). Whether frozen sections or IF are easier is unestablished; ICC-IF shows mainly Golgi localization (HPA: Golgi apparatus supported), and endogenous peroxidase in colonic inflammatory cells can complicate chromogenic interpretation (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
Use compartment-aware controls and matched processing to troubleshoot BLZF1 staining in paraffin sections.
What retrieval should I use when BLZF1 staining is weak in paraffin sections?
Start with heat-mediated Tris-EDTA at pH 8.0 for paraffin-section IHC (datasheet A09997). The selected human brain image used high-pressure, high-temperature retrieval in that buffer and an antibody dilution of 1:100 overnight at 4°C (caption A09997). If staining remains weak, compare a second retrieval condition on adjacent sections while holding antibody dilution, detection, and imaging constant (standard IHC practice). Judge recovery by cellular distribution as well as intensity: BLZF1 is mainly Golgi-localized, with additional nucleoplasmic localization reported, so diffuse staining alone is a poor endpoint (HPA subcellular). Include a no-primary control to reveal detection-related signal (standard IHC practice).
Could fixation explain variable BLZF1 staining across my tissue sections?
Target-specific BLZF1 sensitivity to fixative type or fixation duration is unknown from the supplied evidence; the selected paraffin-section caption does not state a fixative (caption A09997). Record fixative, fixation duration, processing history, and section age for each specimen before comparing staining intensity (standard IHC practice). Process comparison sections together and apply the same Tris-EDTA retrieval at pH 8.0, antibody dilution, and detection conditions (datasheet A09997; standard IHC practice). If a batch loses signal, examine a previously responsive control section processed alongside it before attributing the difference to BLZF1 biology (standard IHC practice). Avoid assigning any particular fixation effect to BLZF1 without a controlled comparison (standard IHC practice).
Should BLZF1 appear in the Golgi, nucleus, or diffuse cytoplasm?
Expect a Golgi-associated pattern as the strongest subcellular reference, with nucleoplasmic signal also supported (HPA subcellular). UniProt lists Golgi membrane localization and notes nuclear detection upon heterologous expression, while its subcellular note says BLZF1 was not detected in the cytoplasm (UniProt Q9H2G9 subcellular). Tissue IHC nevertheless has a general nuclear and cytoplasmic profile with only Supported reliability and medium RNA–staining consistency (HPA tissue IHC). In chromogenic sections, compare signal distribution with intact cellular morphology and a matched negative control before calling broad cytoplasmic color specific (standard IHC practice). Score nuclear and cytoplasmic staining separately when the pattern varies among specimens (standard IHC practice).
How could BLZF1 isoforms or epitope placement affect an IHC result?
BLZF1 has 2 annotated isoforms, but the supplied product caption does not identify the antibody epitope or establish isoform selectivity (UniProt Q9H2G9 isoforms; caption A09997). The protein has no annotated transmembrane segment and has recorded phosphorylation sites at residues 15, 49, 348, and 353 (UniProt Q9H2G9 topology and modified residues). These annotations alone do not show whether processing or phosphorylation changes this antibody’s staining (standard IHC interpretation). If staining differs by specimen, check the antibody’s epitope documentation and compare independently processed sections before assigning an isoform-specific pattern (standard IHC practice). Treat peptide pre-absorption as a specificity check for the selected image, not proof that the reagent distinguishes isoforms (caption A09997; standard IHC interpretation).
How should I check a BLZF1 IHC pattern by multiplex IF?
Use IF as a separate validation experiment: multiplex BLZF1 with a Golgi marker and a cell-type marker appropriate to the specimen, such as an epithelial marker for glandular cells (HPA subcellular; HPA tissue IHC; standard IF practice). For the documented high-staining colon glandular cells, choose spectrally separated fluorophores and compare unstained tissue autofluorescence before interpreting overlap (HPA tissue IHC; standard IF practice). Permeabilize sufficiently to expose intracellular epitopes, but optimize detergent conditions because the antibody epitope and its membrane-facing side are not supplied (UniProt Q9H2G9 topology; standard IF practice). Include single-stain and no-primary controls to assess bleed-through and background, then compare Golgi and nucleoplasmic patterns with the chromogenic section (HPA subcellular; standard IF practice).
What should I check when BLZF1 IHC shows widespread brown background?
Run a no-primary section through the full detection sequence to separate reagent or tissue background from antibody-dependent staining (standard IHC practice). In chromogenic IHC, check the peroxidase block before DAB development and inspect for endogenous pigment that could resemble brown reaction product (standard IHC practice). Review antibody concentration against the selected image’s 1:100 overnight condition at 4°C, while treating that image as one tested condition rather than a universal optimum (caption A09997). Evaluate whether signal respects recognizable Golgi or nuclear compartments, because BLZF1 has supported Golgi and nucleoplasmic localization (HPA subcellular). Compare tissue edges and damaged areas with intact interior regions before interpreting uniform color as expression (standard IHC practice).
How should I quantify BLZF1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the anatomical compartment and cell population before scoring, and apply the same threshold to all sections within a comparison (standard IHC practice). For cellular staining, report percent positive cells and an H-score using intensity categories of 0–3; record nuclear and cytoplasmic scores separately when both occur (standard IHC practice; HPA tissue IHC). If measuring stained profiles in an area rather than individual cells, report their density per mm² of evaluable tissue (standard IHC practice). Normalize counts to the relevant cell population or analyzable tissue area, excluding folds, necrosis, and edge artifacts (standard IHC practice). Keep retrieval, exposure, detection, and scoring rules matched across batches (standard IHC practice).
How can I distinguish credible BLZF1 signal from tissue artifacts?
Give greatest weight to reproducible staining in intact cells with a plausible Golgi-associated or nucleoplasmic distribution (HPA subcellular; standard IHC practice). High HPA tissue staining is reported in cerebral cortex neuropil, colon and duodenal glandular cells, and kidney tubular cells, while adipocytes are reported as not detected (HPA tissue IHC). These references help select comparisons, but the HPA tissue profile has Supported reliability with medium consistency against RNA data (HPA tissue IHC). Inspect section edges, necrotic regions, endogenous pigment, and residual peroxidase activity before calling unexpected brown signal positive (standard IHC practice). The selected brain image includes an immunogen-peptide pre-absorption control, which supports that image’s antibody-dependent staining without resolving every tissue or compartment pattern (caption A09997; standard IHC interpretation).
Boster reagents
Best BLZF1 / Golgin-45 IHC Antibodies
The catalog lists human and mouse IHC/IF options for BLZF1; A09997 has an IHC image from paraffin-embedded human brain, while no IF image is supplied (catalog applications/reactivity; A09997 IHC caption).
Real IHC data
Anti-BLZF1/Golgin 45 Antibody
Cat # A09997
Only A09997 renders a card: its IHC image shows paraffin-embedded human brain at 1:100, with Tris-EDTA pH 8.0 retrieval and a peptide-preabsorption control (A09997 IHC caption). Its catalog entry lists human and mouse reactivity and IHC, IF, and ICC applications; the supplied image documents human brain IHC only (A09997 catalog; A09997 IHC caption).
Which to pick: For tissue IHC, choose A09997 because its rabbit polyclonal antibody has a human brain paraffin-section image; the fixative is unreported (A09997 catalog; A09997 IHC caption). For IF/ICC, A09997 lists both applications and human/mouse reactivity, with a 1:200–1:1000 dilution range, although no IF image is supplied (A09997 catalog). For rat work, A09997-2 lists rat reactivity and IHC/IF applications, but has no supplied IHC or IF image; A09997-1 lists human/mouse IHC and likewise has no supplied image (catalog applications/reactivity; image alts).
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/cytoplasmic tissue staining (HPA tissue IHC); Golgi membrane annotated (UniProt). 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); Caveat: Tissue staining differs from the annotated Golgi location (HPA tissue IHC) (UniProt); Isoform / epitope: 2 isoforms; epitope differences unspecified (UniProt); Regulation: Low tissue RNA 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.