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BTF3L4 / Transcription factor BTF3 homolog 4 · IHC design guide
Design Immunohistochemistry for BTF3L4
Plan BTF3L4 paraffin IHC around general cytoplasmic staining, with high signal in cerebral cortex neurons and skeletal muscle myocytes (HPA tissue IHC). The catalog antibody’s IHC dilution range is 1:100–1:300 (datasheet).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
BTF3L4 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 | General cytoplasmic staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic; high in neurons and myocytes (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Cerebral cortex+4 more · see all | |
| Negative control | Bone marrow+1 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 | Antibody staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | No expression regulator annotated (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope coverage unreported (UniProt; datasheet) |
Section 1
Recommended BTF3L4 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet) with one published BTF3L4 liver staining method (PMC10899873).
| Sample | Paraffin-embedded human brain tissue; fixative not specified (datasheet A15784) |
| Fixation | Image fixative and duration unreported (datasheet A15784); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| 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-BTF3L4, 1:100 - 1:300 (datasheet A15784) |
| 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 | BTF3L4-positive staining in neuronal cells of cerebral cortex (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval rule); the published method leaves its retrieval buffer unspecified (PMC10899873).
Section 2
What Is the Expected BTF3L4 Staining Pattern?
In paraffin-section IHC, expect a general cytoplasmic BTF3L4 pattern, with High staining reported in cerebral-cortex neuronal cells and skeletal-muscle myocytes (HPA: tissue IHC). ICC-IF also places BTF3L4 in the nucleoplasm and cytosol (HPA: approved subcellular locations). UniProt annotates no transmembrane segment (UniProt Q96K17: topology). Tissue IHC reliability is Approved, with medium staining–RNA consistency and external verification pending (HPA: tissue IHC).
| Clear cytoplasmic staining in cortical neurons or skeletal-muscle myocytes, with little surrounding haze (HPA: High in both cell types). | This matches the reported IHC distribution (HPA: general cytoplasmic expression). Score the named cells and their intensity separately from the rest of the section; a clean cell-shaped deposit is easier to interpret than widespread color (general IHC practice). |
| Predominantly nuclear or membrane-rim staining in an IHC section, with little cytoplasmic signal. | Recheck the IHC pattern and assay controls: HPA describes general cytoplasmic tissue staining, while ICC-IF also supports nucleoplasm (HPA: tissue IHC; HPA: approved ICC-IF locations). Nuclear signal alone is therefore not proof of artefact; membrane-rim signal has no support from the supplied localization or topology record (UniProt Q96K17: no transmembrane segment). |
| Strong staining in bone-marrow hematopoietic cells or soft-tissue fibroblasts. | Those cells were Not detected in HPA tissue IHC (HPA: bone marrow; HPA: soft tissue). Investigate cross-reactivity or detection-derived color before assigning the deposit to BTF3L4; a reference image is a comparison, not proof that every specimen must be negative (general IHC practice). |
| Uniform brown haze across cells, stroma, and blank spaces, obscuring cell boundaries. | A field-wide deposit does not resemble HPA's cell-associated cytoplasmic pattern (HPA: tissue IHC). Check reagent background and nonspecific detection with appropriate controls; interpret cellular intensity only after the haze is reduced (general IHC practice). |
| No convincing cytoplasmic staining in cortical neurons or skeletal-muscle myocytes. | These are reported High examples, so absent signal warrants an assay check (HPA: tissue IHC). First compare tissue preservation and a run control, then review antibody use and detection steps (general IHC practice). A single negative section cannot establish absent BTF3L4 expression. |
💡Expected BTF3L4 appearanceCall an IHC result concordant when cortical neurons or skeletal-muscle myocytes show distinct, preferably High, cytoplasmic staining (HPA: tissue IHC); diffuse haze or strong color in HPA Not detected cell types calls for a false-positive check (HPA: bone marrow and soft tissue; general IHC practice).
| What should IF/ICC show? | HPA reports approved nucleoplasm and cytosol localization in ICC-IF (HPA: subcellular). That result provides context for a nuclear IHC observation, but tissue IHC is summarized as generally cytoplasmic (HPA: tissue IHC). Judge each assay against its own reported pattern. |
| Choice of tissue and cell population | Cortical neurons and skeletal-muscle myocytes provide High reference staining; adrenal or appendix glandular cells are Medium (HPA: tissue IHC). Parathyroid glandular cells and liver cholangiocytes are Low (HPA: tissue IHC). Compare like cell types when assessing intensity. |
| Antibody validation | HPA067026 is IHC Approved and ICC Approved; HPA074056 is ICC Approved with no IHC status listed (HPA: antibody validation). The tissue profile is Approved, with medium staining–RNA consistency and external verification pending (HPA: tissue IHC). Match validation to the assay when weighing unexpected staining. |
| Isoforms and epitope coverage | UniProt lists 3 BTF3L4 isoforms (UniProt Q96K17: alternative splicing). The supplied record does not map the catalog antibody's epitope to those isoforms; an unusual pattern cannot be attributed to isoform selection from this evidence alone. |
| Protein architecture | The annotated chain spans residues 1–158, with no signal peptide, propeptide, or transmembrane segment (UniProt Q96K17: processing and topology). These annotations offer no basis for expecting a membrane-rim IHC pattern; they do not establish fixation sensitivity or antibody epitope accessibility. |
| Situation | Likely cause | Next action |
|---|---|---|
| A known High reference cell population is unstained (HPA: cortical neurons or skeletal-muscle myocytes). | An unsuccessful stain, a section-quality issue, or weak detection can produce an apparent negative (general IHC practice). The supplied sources identify no BTF3L4-specific fixation effect. | Confirm the run control, tissue morphology, antibody application, and detection sequence; repeat with a suitable reference section if needed (general IHC practice). |
| The whole IHC section has diffuse color that masks cytoplasm. | Nonspecific reagent binding or inadequate washing can raise background (general IHC practice). HPA instead reports general cytoplasmic expression (HPA: tissue IHC). | Inspect a no-primary control, blocking, wash steps, and detection reagent exposure; adjust the general workflow only after locating the source of background (general IHC practice). |
| IHC staining appears nuclear only. | This differs from the general tissue-IHC profile, though nucleoplasm is an approved ICC-IF location (HPA: tissue IHC; HPA: subcellular). The compartment difference alone cannot identify the cause. | Check cell morphology and staining controls, then compare cytoplasmic signal in a High reference population before assigning the IHC result to BTF3L4 (HPA: tissue IHC; general IHC practice). |
| Bone-marrow hematopoietic cells or soft-tissue fibroblasts stain strongly. | HPA reports Not detected staining in these populations (HPA: tissue IHC). Cross-reactivity or detection background is possible, but the image alone cannot distinguish them. | Compare a no-primary control and the expected positive cells on a reference section; inspect whether color follows cell boundaries or occurs outside them (general IHC practice). |
| Granular chromogen appears in many cells regardless of the expected cell pattern. | If an HRP detection system is used, endogenous peroxidase activity can generate color unrelated to primary-antibody binding (general IHC practice). | Use a no-primary control and review the endogenous-peroxidase blocking step for that detection system; reassess only well-defined cellular staining (general IHC practice). |
| Color forms a membrane rim while the cytoplasm remains faint. | A membrane pattern is unsupported by HPA's general cytoplasmic tissue profile and UniProt's lack of a transmembrane segment (HPA: tissue IHC; UniProt Q96K17: topology). | Check section edges and reagent background, then compare an expected positive cell population and a no-primary control before treating the rim as specific (general IHC practice; HPA: tissue IHC). |
Experiment evidence
IHC & IF images showing BTF3L4 staining
A15784 · IHCImmunohistochemistry analysis of paraffin-embedded human brain tissue, using BTF3L4 Antibody. The picture on the right is blocked with the s
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000134717-BTF3L4/tissue/cerebral+cortex#imid_18236111
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000134717-BTF3L4/tissue/colon#imid_18236037
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000134717-BTF3L4/tissue/liver#imid_18236054
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000134717-BTF3L4/tissue/kidney#imid_18236047
HPA · ICC-IF SH-SY5YHuman Protein Atlas immunofluorescence, SH-SY5Y, antibody HPA067026; green = BTF3L4, blue = nucleus, red = microtubules.
HPA · ICC-IF SH-SY5YHuman Protein Atlas immunofluorescence, SH-SY5Y, antibody HPA067026; green = BTF3L4, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for BTF3L4 IHC & IF
🧪Run cerebral cortex first and look for staining in neuronal cells (HPA: High in cerebral cortex neuronal cells). Use bone marrow hematopoietic cells as the biological negative (HPA: Not detected); within the cortex section, treat unstained cells as background comparators without assuming a particular non-neuronal cell type is negative.
Positive control tissue: Cerebral cortex (Neuronal cells, HPA High)
Negative control tissue: Bone marrow (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show BTF3L4 in SH-SY5Y, U-251MG, U2OS, HBEC3-KT, with annotated localisation: Nucleoplasm (approved), Cytosol (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and an isotype control matched to the primary antibody’s host species and clonality (standard IHC practice). If available, use BTF3L4 knockout material as a biological specificity control, or perform a peptide block as shown for the catalog antibody (A15784 tissue-IHC caption). For cortex, block endogenous peroxidase for chromogenic detection and check background pigment; for IF, assess tissue autofluorescence (standard IHC/IF practice).
⚠️Feasibility: The selected paraffin-section brain IHC caption does not report a fixative, and the supplied evidence reports no BTF3L4-specific fixation window or antigen-retrieval dependence (A15784 tissue-IHC caption). There is no supplied comparison showing that frozen sections or IF are easier; HPA does show ICC-IF images and reports nucleoplasm and cytosol localization (HPA subcellular). In brain sections, background pigment can complicate chromogenic scoring and autofluorescence can complicate IF scoring (standard IHC/IF practice).
HPA tissue IHC evidence for BTF3L4
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium 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 |
|---|---|---|---|---|
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced BTF3L4 IHC Tips
Use matched paraffin sections and compartment-aware controls to troubleshoot BTF3L4 staining; the selected tissue image reports paraffin embedding but leaves fixation unreported (A15784 caption).
How should I optimize retrieval when BTF3L4 staining is weak?
Start with citrate buffer at pH 6.0 for heat-induced retrieval at 95–98 °C for 20 min (page retrieval setting). The selected A15784 image shows paraffin-embedded human brain but reports neither fixative nor retrieval conditions, so treat this schedule as a starting point rather than image-matched validation (A15784 caption). Compare adjacent sections with and without retrieval while keeping antibody incubation and chromogen development identical, then assess neuronal signal and background (HPA: high cerebral-cortex neuronal staining; standard IHC practice). If staining remains weak, test a longer citrate exposure before an alternative buffer, watching for tissue detachment and loss of cellular detail (standard IHC practice).
Could fixation explain variable BTF3L4 staining between paraffin blocks?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not identify its fixative, and no BTF3L4 fixation comparison is supplied (A15784 caption). Record fixative, fixation duration, processing history and block age for each specimen before attributing intensity differences to biology (standard IHC practice). If matched material is available, compare sections fixed in 10% neutral buffered formalin for 6 and 24 hours, using the same citrate pH 6.0 retrieval and staining run (standard IHC practice; page retrieval setting). Interpret any difference as specific to that comparison until replicated with additional specimens, because fixation and tissue composition can both affect chromogenic intensity (standard IHC practice).
Should BTF3L4 staining be cytoplasmic or nuclear in tissue?
Expect predominantly cytoplasmic tissue staining as the initial IHC reference, while allowing a separately scored nuclear component because cellular imaging places BTF3L4 in both cytosol and nucleoplasm (HPA tissue IHC: general cytoplasmic expression; HPA subcellular: approved cytosol and nucleoplasm). UniProt supplies no subcellular annotation and records no transmembrane segment, so membrane-restricted outlines lack support from this record (UniProt Q96K17). Score nuclear and cytoplasmic signal independently within identified cells, using identical DAB development across comparison sections (standard IHC practice). If staining shifts entirely to one compartment between runs, review retrieval, counterstain and control slides before treating the shift as biological (standard IHC practice).
How can I assess whether the antibody detects all BTF3L4 isoforms?
BTF3L4 has 3 annotated isoforms, and its NAC-A/B domain spans residues 33–98 (UniProt Q96K17). Without the antibody’s mapped epitope and isoform sequences, staining cannot establish which isoforms contribute to an IHC signal (UniProt Q96K17; standard IHC interpretation). Obtain the immunogen or epitope sequence, align it to each isoform, and check whether fixation and citrate pH 6.0 retrieval preserve staining in matched sections (standard IHC practice; page retrieval setting). The A15784 brain image includes a synthesized-peptide block, which supports peptide-competitive staining in that image but does not resolve isoform coverage or prove specificity against unrelated proteins (A15784 caption; standard IHC interpretation).
How should I adapt the tissue finding for BTF3L4 multiplex IF?
For a separate IF experiment, pair BTF3L4 with a neuronal marker when testing cerebral cortex, where neuronal cells show high tissue staining (HPA tissue IHC: cerebral-cortex neuronal cells, high). Select spectrally separated fluorophores after inspecting unstained tissue for autofluorescence, and include single-color controls to assess bleed-through (standard IF practice). Because cellular imaging places BTF3L4 in cytosol and nucleoplasm, use permeabilisation that permits access to intracellular epitopes and check whether nuclear and cytosolic patterns remain visible (HPA subcellular: approved cytosol and nucleoplasm; standard IF practice). Optimize fixation and permeabilisation directly for IF; the A15784 paraffin-tissue caption provides no IF fixation conditions (A15784 caption).
What controls help distinguish BTF3L4 signal from chromogenic background?
Run a no-primary control and inspect pigment, endogenous peroxidase and diffuse DAB deposition before assigning positive cells (standard IHC practice). Apply a peroxidase block and an appropriate protein block as general chromogenic workflow steps, then keep DAB development time identical across compared sections (standard IHC practice). The selected A15784 paraffin-brain image includes a synthesized-peptide-blocked companion image, providing a useful competition reference for that antibody and specimen (A15784 caption). Compare cellular staining with the reported high neuronal staining in cerebral cortex and undetected hematopoietic cells in bone marrow, while recognizing that those HPA observations are context rather than absolute negative controls for every specimen (HPA tissue IHC).
How should I quantify BTF3L4 staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define cell type, compartment, staining threshold and analysis regions before comparing slides, because the tissue profile reports general cytoplasmic expression while cellular imaging also supports nucleoplasmic signal (HPA tissue IHC; HPA subcellular). For intensity, score each cell from 0–3 and calculate an H-score as the sum of intensity multiplied by the percentage of cells at that intensity, yielding 0–300 (standard IHC practice). Alternatively, report percentage of positive cells or positive-cell density per mm² within a defined tissue compartment (standard IHC practice). Normalize to the relevant cell population or sampled viable tissue area, and compare sections processed in the same staining run with identical thresholds (standard IHC practice).
When should a BTF3L4-positive cell be considered convincing?
A convincing result shows cellular staining in an expected population, such as cerebral-cortex neurons, with a predominantly cytoplasmic tissue pattern and a separately assessed nuclear component (HPA tissue IHC: high neuronal staining and general cytoplasmic expression; HPA subcellular: approved nucleoplasm). Strong membrane-only staining has no support from the recorded topology, which lacks a transmembrane segment (UniProt Q96K17). Examine section edges, necrotic regions and no-primary controls for nonspecific DAB or endogenous enzyme signal before counting positive cells (standard IHC practice). The A15784 peptide-blocked brain image is supporting evidence for its observed signal, but HPA tissue staining has medium RNA concordance and awaits external verification, so corroborate unexpected distributions independently (A15784 caption; HPA tissue IHC: Approved reliability description).
Boster reagents
Best BTF3L4 / Transcription factor BTF3 homolog 4 IHC Antibodies
A15784 has an IHC image from paraffin-embedded human brain tissue (catalog image caption); IF/ICC is listed without an image (catalog applications), and Human and Mouse are listed as reactive species (catalog reactivity).
Real IHC data
Anti-BTF3L4 Antibody
Cat # A15784
A15784 will render with its IHC image of paraffin-embedded human brain tissue and a peptide-blocked comparison (catalog image caption). Its listed applications include IHC, IF and ICC, with Human and Mouse reactivity; no IF image is supplied (catalog applications; catalog reactivity; catalog image list).
Which to pick: For tissue IHC, choose A15784: its own image documents staining in paraffin-embedded human brain tissue, with a peptide-blocked comparison (catalog image caption); the fixative is unreported (catalog image caption). For IF/ICC, A15784 lists both applications and is polyclonal, though no IF image is supplied (catalog applications; catalog dilution data; catalog image list). For work involving Human and Mouse samples, A15784 lists reactivity with both species; the supplied tissue image documents Human only (catalog reactivity; catalog image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q96K17 (BT3L4_HUMAN, Transcription factor BTF3 homolog 4).
- Human Protein Atlas. BTF3L4 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. BTF3L4 subcellular location (ICC-IF): Mainly localized to the nucleoplasm and cytosol..
- Human Protein Atlas. BTF3L4 antibody validation summary (2 antibodies).
- BTF3L4 Overexpression Mediates APAP-induced Liver Injury in Mouse and Cellular Models. Journal of clinical and translational hepatology 2024 — PMC10899873.
- Multi-omics analysis of BTF3L4 as a prognostic and immune biomarker in hepatocellular carcinoma. Translational cancer research 2026 — PMC12971597.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16710414 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for BTF3L4 (Transcription factor BTF3 homolog 4). Expected localisation General cytoplasmic staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A15784 at 1:100 - 1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Cerebral cortex (Neuronal cells); negative Bone marrow. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human brain tissue; fixative not specified (datasheet A15784); 2. Fixation: Image fixative and duration unreported (datasheet A15784); 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-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 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-BTF3L4, 1:100 - 1:300 (datasheet A15784); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Antibody staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope coverage unreported (UniProt; datasheet); Regulation: No expression regulator annotated (UniProt). Evidence: UniProt Q96K17, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm and cytosol., Datasheet A15784, PMC10899873. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/BTF3L4/immunohistochemistry-ihc-guide.