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ABCF1 / ATP-binding cassette sub-family F member 1 · IHC design guide
Design Immunohistochemistry for ABCF1
Plan ABCF1 chromogenic IHC in paraffin sections using the cytoplasmic pattern reported across most tissues (HPA tissue IHC). Colon glandular cells show high staining, while adipocytes are not detected; verify antibody specificity before interpreting either result (HPA tissue IHC; datasheet).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ABCF1 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 IHC; nuclear location is annotated (HPA tissue IHC; UniProt) | |
| Staining pattern | Cytoplasmic staining in most tissues, including glandular cells (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Breast+4 more · see all | |
| Negative control | Adipose tissue |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Catalog text describes phospho-GluR1; verify ABCF1 specificity (datasheet) | |
| Regulation | Ubiquitous expression (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unspecified (UniProt; datasheet) |
Section 1
Recommended ABCF1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by one published ABCF1 protocol for paraffin-embedded bladder tissue (PMC10022215: IHC methods).
| Sample | Paraffin-embedded human colon tissue; fixative not specified (datasheet M05358) |
| Fixation | Image fixative and duration unreported (datasheet M05358); 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 Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear 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 monoclonal (clone FGE-1) anti-ABCF1, 1:50 (datasheet M05358) |
| 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 | ABCF1-positive staining in glandular cells of breast (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 HIER at 95–98 °C for 20 min (page antigen retrieval); the bladder study does not report retrieval (PMC10022215: IHC methods).
Section 2
What Is the Expected ABCF1 Staining Pattern?
ABCF1 should show predominantly cytoplasmic staining in most tissues, with strong signal in several epithelial and other cell populations (HPA: tissue IHC profile; HPA: High in breast glandular, esophageal squamous epithelial, lymph node germinal center, and placental trophoblastic cells). Nuclear or nuclear-envelope signal is biologically plausible, but a membrane-dominant pattern is unexpected because ABCF1 has no transmembrane segment (UniProt Q8NE71: localization and topology). Tissue IHC reliability is Approved, with medium consistency against RNA and external verification pending (HPA: tissue IHC reliability).
| Cytoplasmic stain in breast or colon glandular cells, with clear cell outlines and limited background (HPA: High in both cell populations). | This fits the reported tissue pattern (HPA: cytoplasmic expression in most tissues). Judge the stain in its cellular context: glandular signal is useful, while intensity across every cell need not be uniform because the HPA profile reports cell-specific levels (HPA: tissue IHC levels). |
| Strong staining confined to plasma membranes, with little cytoplasmic signal (HPA: cytoplasmic tissue profile). | Investigate an off-target or detection artefact before calling this ABCF1: the dominant compartment conflicts with tissue IHC and with a protein lacking a transmembrane segment (HPA: tissue IHC profile; UniProt Q8NE71: topology). Nuclear or nuclear-envelope signal alone deserves separate review because those locations are listed by UniProt (UniProt Q8NE71: localization). |
| Prominent stain in adipocytes while adjacent expected-positive glandular cells remain unstained (HPA: adipocytes Not detected; glandular cells High in breast and colon). | Suspect cross-reactivity or endogenous detection activity and review controls (general IHC practice). HPA's adipocyte result is an observed IHC category, not proof that every adipocyte must be blank in every specimen (HPA: adipocytes Not detected; tissue IHC reliability). |
| Haze across tissue, empty spaces, and cell-free regions obscures cell boundaries (general IHC practice). | This distribution is diffuse background rather than interpretable cell-localized ABCF1 signal (general IHC practice; HPA: cytoplasmic tissue profile). Review the detection control, blocking, washing, and chromogen development before scoring intensity (general IHC practice). |
| No visible stain in a technically intact section containing breast glandular cells (HPA: High in breast glandular cells). | Treat the run as unresolved, not as evidence of absent ABCF1: HPA reports strong staining in that cell population, subject to its Approved reliability and pending external verification (HPA: breast glandular cells High; tissue IHC reliability). Check tissue identity and assay controls first (general IHC practice). |
💡Expected ABCF1 appearanceCall a positive result when cytoplasmic staining is clear in an expected-positive cell population, such as breast glandular cells reported High by HPA; isolated membrane-dominant or cell-free staining should prompt artefact review (HPA: cytoplasmic profile and breast glandular cells High; UniProt Q8NE71: topology; general IHC practice).
| Tissue and cell selection (HPA: tissue IHC profile). | Breast, colon, duodenum, endometrium, and gallbladder glandular cells are reported High; hepatocytes and lung alveolar cells are Low, and adipocytes are Not detected (HPA: tissue IHC levels). Compare the intended cell population, not just the organ name (general IHC practice). |
| Compartment and topology (UniProt Q8NE71: localization and topology). | UniProt lists cytoplasm, nucleoplasm, and nuclear envelope, with no transmembrane segment (UniProt Q8NE71: localization and topology). HPA tissue IHC describes predominantly cytoplasmic expression, so nuclear signal merits context and membrane-dominant signal merits investigation (HPA: tissue IHC profile; general IHC practice). |
| Isoform coverage (UniProt Q8NE71: isoforms). | Two isoforms are listed, but the supplied record does not map the IHC antibody's epitope or establish isoform-specific staining (UniProt Q8NE71: isoforms; supplied antibody record). Do not assign a slide pattern to either isoform from these data. |
| Antibody evidence (HPA: HPA017578 validation). | HPA lists HPA017578 as IHC Approved and ICC Enhanced; the tissue profile has medium antibody–RNA consistency and awaits external verification (HPA: antibody validation and tissue IHC reliability). These ratings support cautious comparison with the reported pattern, not proof of specificity in a new specimen. |
| IF/ICC Q&A: What should the separate IF/ICC guide expect? (HPA: subcellular ICC-IF). | Cytosolic localization is the enhanced main location in HPA ICC-IF, with images listed for A-431, U-251MG, and U2OS contexts (HPA: subcellular ICC-IF). Use that as a localization cross-check; it does not set an IHC-P protocol or guarantee identical intensity across methods (general assay interpretation). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected-positive glandular cells are blank (HPA: breast and colon glandular cells High). | Section identity, retrieval, antibody incubation, or detection may need review (general IHC practice); target-specific fixation sensitivity is unreported in the supplied evidence. | Confirm the cell population and run controls, then check the validated IHC-P workflow's retrieval, incubation, and detection steps (general IHC practice). |
| Stain is mainly at cell borders (HPA: cytoplasmic tissue profile). | A membrane-dominant result conflicts with reported localization and absent transmembrane topology; its precise cause cannot be assigned from the pattern alone (HPA: tissue IHC profile; UniProt Q8NE71: topology). | Inspect morphology and detection controls, then repeat with an appropriate antibody control if available (general IHC practice). |
| Adipocytes stain strongly (HPA: adipocytes Not detected). | Off-target binding or endogenous detection activity is possible (general IHC practice); HPA's category alone cannot identify the mechanism (HPA: adipocytes Not detected). | Check a primary-omission detection control and review blocking and chromogen development (general IHC practice). |
| Background masks cytoplasmic detail (HPA: expected cytoplasmic tissue profile). | Nonspecific reagent retention or excessive development can obscure cell-specific signal (general IHC practice). | Review blocking, washes, and development time alongside a detection control; score only resolved cellular staining (general IHC practice). |
| Only nuclear staining is apparent (UniProt Q8NE71: nucleoplasm and nuclear envelope). | Nuclear localization is listed, but a nuclear-only tissue pattern differs from HPA's predominantly cytoplasmic IHC profile (UniProt Q8NE71: localization; HPA: tissue IHC profile). | Verify the compartment with the counterstain and compare with a known-positive section before interpreting the result (general IHC practice; HPA: tissue IHC profile). |
| A Low cell population looks weaker than a High one (HPA: tissue IHC levels). | Cell-specific staining levels differ in the reference profile; this comparison alone does not show assay failure (HPA: hepatocytes Low; breast glandular cells High). | Interpret each identified cell population against its reported level and the same-run controls (HPA: tissue IHC levels; general IHC practice). |
Experiment evidence
IHC & IF images showing ABCF1 staining
M05358 · IHCImmunohistochemical analysis of paraffin-embedded human colon, using ABCF1 Antibody.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000204574-ABCF1/tissue/cerebral+cortex#imid_4610784
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000204574-ABCF1/tissue/colon#imid_4610726
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000204574-ABCF1/tissue/liver#imid_4610719
HPA · GallbladderHuman Protein Atlas IHC, Gallbladder. Source: https://www.proteinatlas.org/ENSG00000204574-ABCF1/tissue/gallbladder#imid_4610695
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA017578; green = ABCF1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA017578; green = ABCF1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ABCF1 IHC & IF
🧪Run colon first: glandular cells should stain strongly (HPA: High in colon glandular cells; selected M05358 caption: paraffin-embedded human colon). Use adipose tissue as the negative comparison (HPA: Not detected in adipocytes); cells without specific staining on the colon slide should show only background signal, without assuming any particular colon cell type is ABCF1-negative.
Positive control tissue: Breast (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ABCF1 in A-431, U-251MG, U2OS, U2OS, siRNA 2 (10x), U2OS, scrambled (10x), with annotated localisation: Cytosol (enhanced) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, an isotype control matched to the primary antibody’s host species and antibody class, and an ABCF1 knockout specimen as a biological negative. For chromogenic colon IHC, quench endogenous peroxidase and check for endogenous biotin if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported in the supplied evidence, and the fixative is unreported in the selected M05358 paraffin-section caption (selected M05358 caption: fixative not stated). Antigen-retrieval dependence is also unreported; optimize retrieval with the IHC-validated antibody rather than inferring it from staining patterns (standard IHC practice). The supplied evidence does not establish whether frozen sections or IF are easier, or identify a colon-specific artefact; HPA reports cytosolic ICC-IF localisation (HPA: Cytosol, enhanced).
HPA tissue IHC evidence for ABCF1
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 |
|---|---|---|---|---|
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | High | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | 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 ABCF1 IHC Tips
Troubleshoot ABCF1 staining in paraffin sections by checking retrieval, cellular distribution, controls, and scoring against reported tissue patterns (HPA tissue IHC; UniProt Q8NE71).
How should I optimize retrieval when ABCF1 staining is weak in paraffin sections?
Start with Tris-EDTA at pH 9.0 for heat-induced retrieval at 95–98 °C for 20 min (page retrieval setting). The catalog antibody has a paraffin-section image from human colon, but its caption does not report the fixative or retrieval method (M05358 tissue-IHC caption). If staining remains weak, test citrate at pH 6.0 as a fallback on matched sections while holding detection conditions constant (standard IHC optimization). Compare glandular cells with an omission control: colon glandular staining is reported as high, so loss of that signal can help identify an inadequate retrieval condition (HPA tissue IHC).
Could fixation explain weak ABCF1 staining in my paraffin sections?
Target-specific fixation sensitivity is unknown: the selected human-colon image identifies a paraffin section but does not state its fixative (M05358 tissue-IHC caption). For a fixation comparison, record the fixative and duration, process matched samples together, and change only one variable at a time (standard IHC practice). A pilot using 10% neutral buffered formalin with a consistent fixation duration is a conventional starting point for new paraffin specimens, not a reported ABCF1 condition (standard IHC practice). Judge each condition against preserved morphology, a no-primary control, and staining in an expected cell population such as colon glandular cells (standard IHC practice; HPA tissue IHC).
Which cellular compartments should show convincing ABCF1 staining?
Expect predominantly cytoplasmic staining in tissue sections, since that is the reported pattern across most tissues (HPA tissue IHC). Cytosol is the enhanced subcellular location, while cytoplasm, nucleoplasm, and nuclear envelope are also annotated for ABCF1 (HPA subcellular; UniProt Q8NE71). Score nuclear signal separately from cytoplasmic signal and check it against a no-primary control before interpreting a change in distribution (standard IHC practice). A strong, continuous cell-surface outline is harder to reconcile with a protein lacking a transmembrane segment, so inspect such staining for edge effects or detection background (UniProt Q8NE71 topology; standard IHC practice).
Can this stain distinguish ABCF1 isoforms or phosphorylation states?
ABCF1 has two annotated isoforms, but the supplied catalog image does not identify the antibody epitope or establish isoform selectivity (UniProt Q8NE71 isoforms; M05358 tissue-IHC caption). Map the immunogen to each isoform before making an isoform-specific claim, and use an independently validated reagent if that distinction matters (standard antibody-validation practice). Isoform 2 associates with ribosomes through both ABC transporter domains and interacts with EIF2S1 through its N-terminus (UniProt Q8NE71 subunit). Several phosphorylated residues are annotated, including positions 22, 24, 105, 108, and 109; ordinary chromogenic staining cannot assign a phosphorylation state without a validated modification-specific epitope (UniProt Q8NE71 modified residues; standard IHC interpretation).
How should I check ABCF1 localisation in a multiplex IF experiment?
For an IF experiment, pair ABCF1 with an epithelial marker such as cytokeratin when evaluating colon glandular cells, an expected high-staining population in tissue IHC (HPA tissue IHC; standard multiplex IF practice). Put the weaker signal in a bright, spectrally separated channel and consider a far-red fluorophore when tissue autofluorescence interferes with shorter wavelengths (standard IF practice). Because ABCF1 is annotated in cytoplasm and nucleus and has no transmembrane segment, trial a mild permeabilisation condition such as 0.1% Triton X-100 for 5 min to access intracellular epitopes (UniProt Q8NE71; standard IF practice). Include single-stain and no-primary controls, and optimize IF fixation independently because no antibody-specific IF fixation condition is supplied (standard IF practice).
What should I check if chromogenic staining is diffuse or widespread?
Run a no-primary section alongside the test section to identify signal from the secondary reagent or detection system (standard IHC practice). For peroxidase-based detection, a 3% hydrogen peroxide block for 10 min and careful control of DAB development are general workflow steps, not ABCF1-specific validation (standard IHC practice). Titrate the catalog antibody from a documented starting condition rather than assuming that stronger chromogen deposition reflects more ABCF1 (standard IHC optimization). Inspect section folds, edges, and damaged areas separately; compare intact glandular cells with adipocytes, which are reported as high and not detected, respectively (standard IHC practice; HPA tissue IHC).
How should I score ABCF1 across heterogeneous tissue regions? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring: cytoplasmic expression is reported in most tissues, while colon glandular cells show high staining (HPA tissue IHC). For an H-score, assign intensity 0–3 and multiply each intensity by its percentage of viable target cells, yielding 0–300 (standard IHC scoring). Report the percentage of positive cells and the number of viable target cells assessed alongside the score (standard IHC quantification). Normalize comparisons to the same cell population or measured viable epithelial area, and exclude necrotic or folded regions using the same rules for every section (standard IHC quantification).
How can I separate genuine ABCF1 signal from staining artefact?
A plausible positive pattern is cytoplasmic staining in intact colon glandular cells, which are reported to stain highly, with appropriate control sections remaining clean (HPA tissue IHC; standard IHC practice). Adipocytes are reported as not detected and can provide a tissue-context check when present in the evaluated material (HPA tissue IHC). Investigate staining concentrated at section edges, folds, or necrotic areas, or staining that persists without primary antibody, as potential artefact (standard IHC practice). Predominantly sharp cell-surface staining warrants scrutiny because ABCF1 has no transmembrane segment; isolated nuclear staining needs control-based assessment because nucleoplasm and nuclear envelope are annotated locations (UniProt Q8NE71 topology and subcellular location; standard IHC interpretation).
Boster reagents
Best ABCF1 / ATP-binding cassette sub-family F member 1 IHC Antibodies
Anti-ABCF1 M05358 has an IHC image from paraffin-embedded human colon (IHC image caption). IF/ICC is listed without an IF image; human and rat reactivity is listed (catalog applications and reactivity).
Real IHC data
Anti-ABCF1 Rabbit Monoclonal Antibody
Cat # M05358
M05358 will render with an IHC image of paraffin-embedded human colon (IHC image caption). Its listed applications include IHC and IF/ICC, and its listed reactivity is human and rat (catalog applications and reactivity).
Which to pick: For tissue IHC, choose M05358: its image documents paraffin-embedded human colon, but the fixative is unreported (IHC image caption). For IF/ICC, M05358 is listed for those applications, although no IF image is supplied (catalog applications and IF image alts). For work involving human and rat samples, M05358 is a rabbit monoclonal with both species listed as reactive; the supplied IHC image documents human tissue only (catalog title and reactivity; IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8NE71 (ABCF1_HUMAN, ATP-binding cassette sub-family F member 1).
- Human Protein Atlas. ABCF1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. ABCF1 subcellular location (ICC-IF): Localized to the cytosol..
- Human Protein Atlas. ABCF1 antibody validation summary (1 antibodies).
- High expression of ABCF1 is an independent predictor of poor prognosis in bladder cancer. BMC urology 2023 — PMC10022215.
- ABCF1 Regulates dsDNA-induced Immune Responses in Human Airway Epithelial Cells. Frontiers in cellular and infection microbiology 2020 — PMC7525020.
- NTRK2 promotes malignant progression and paclitaxel resistance of lung adenocarcinoma through targeting MYC/ABCF1 axis. Translational oncology 2026 — PMC12813135.
- PubMed PMID:9790762 — UniProt-cited evidence.
- PubMed PMID:14574404 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ABCF1 (ATP-binding cassette sub-family F member 1). Expected localisation Cytoplasmic in tissue IHC; nuclear location is annotated (HPA tissue IHC; UniProt). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: M05358 at 1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Breast (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colon tissue; fixative not specified (datasheet M05358); 2. Fixation: Image fixative and duration unreported (datasheet M05358); 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 Tris-EDTA buffer, pH 9.0 , 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 monoclonal (clone FGE-1) anti-ABCF1, 1:50 (datasheet M05358); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Catalog text describes phospho-GluR1; verify ABCF1 specificity (datasheet); Isoform / epitope: 2 isoforms; epitope coverage is unspecified (UniProt; datasheet); Regulation: Ubiquitous expression (UniProt). Evidence: UniProt Q8NE71, HPA tissue IHC · Approved, HPA ICC-IF · cytosol., Datasheet M05358, PMC10022215. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ABCF1/immunohistochemistry-ihc-guide.