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ABCF1 / ATP-binding cassette sub-family F member 1 · IHC design guide
Design Immunohistochemistry for ABCF1
Plan chromogenic ABCF1 IHC in paraffin sections with the catalog antibody at a 1:50 starting dilution (datasheet). Use colon glandular cells as a high-staining reference and adipocytes as a not-detected reference (HPA tissue IHC).
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 tissue staining (HPA tissue IHC); nuclear locations annotated (UniProt) | |
| Staining pattern | Cytoplasmic in most tissues; high in several glandular cell types (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 | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image M05358) | |
| Caveat | Catalog text also describes phospho-GluR1; verify specificity (datasheet) | |
| Regulation | Ubiquitous expression; no stated inducer (UniProt) | |
| Isoform / epitope | 2 isoforms; no TM segment; epitope coverage unknown (UniProt) |
Section 1
Recommended ABCF1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with the published bladder tissue protocol (PMC10022215).
| 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 retrieval; UniProt: nucleoplasm). The bladder paper does not specify retrieval (PMC10022215).
Section 2
What Is the Expected ABCF1 Staining Pattern?
ABCF1 staining should be mainly cytoplasmic across most tissues, with strong staining expected in several glandular and epithelial cell types (HPA: tissue IHC). UniProt also lists the nucleoplasm and nuclear envelope, and reports no transmembrane segment (UniProt Q8NE71: localization and topology). Treat the tissue pattern as a guide: HPA rates its IHC evidence Approved, with medium agreement between staining and RNA data and external verification pending (HPA: tissue IHC).
| Clear cytoplasmic staining in breast or colon glandular cells. | This matches HPA's High staining in those cells and its cytoplasmic pattern across most tissues (HPA: tissue IHC). Compare cells within the section before assigning an overall score; a positive field alone does not describe the whole tissue (general IHC practice). |
| Staining is confined to cell borders, with little cytoplasmic signal. | A border-only pattern conflicts with HPA's cytoplasmic IHC profile and ABCF1's lack of a transmembrane segment (HPA: tissue IHC; UniProt Q8NE71: topology). Check morphology and detection controls before calling it ABCF1. Nuclear staining alone also needs caution: UniProt lists nuclear locations (UniProt Q8NE71: localization). |
| Strong staining appears in adipocytes while nearby expected-positive cells are weak. | HPA reports ABCF1 as Not detected in adipocytes, so this result warrants a cross-reactivity or detection-background check (HPA: adipose tissue IHC). It is not proof of either cause; review a reagent control and an independently stained positive tissue (general IHC practice). |
| Color spreads across tissue spaces or obscures cell boundaries. | A diffuse deposit cannot establish the cytoplasmic, cell-specific pattern reported by HPA (HPA: tissue IHC). Inspect reagent controls, blocking and wash performance, and whether chromogen development has overwhelmed cellular detail (general IHC practice). |
| No staining appears in breast glandular cells on an otherwise interpretable section. | HPA reports High staining in breast glandular cells (HPA: breast IHC). First check the positive control and run conditions; a blank result cannot distinguish low antigen from a failed IHC run without those checks (general IHC practice). |
💡Expected ABCF1 appearanceCall a positive result when cytoplasmic staining is readily visible in glandular or epithelial cells reported as High by HPA; border-only deposits, widespread adipocyte staining, or color without cellular detail warrant control review (HPA: tissue IHC; UniProt Q8NE71: topology).
| Compartment and topology | HPA describes cytoplasmic expression in most tissues (HPA: tissue IHC). UniProt also lists nucleoplasm and nuclear envelope, but no transmembrane segment (UniProt Q8NE71: localization and topology). Interpret nuclear signal with controls rather than requiring it in every section. |
| Tissue and cell selection | High-staining examples include duodenal and gallbladder glandular cells, esophageal squamous cells, lymph-node germinal-center cells, and placental trophoblasts (HPA: tissue IHC). Adipocytes are reported Not detected; several other listed cell types are Low (HPA: tissue IHC). |
| Isoforms and antibody scope | UniProt lists two ABCF1 isoforms and assigns ribosome association and translation-initiation functions specifically to isoform 2 (UniProt Q8NE71: isoforms and function). The supplied IHC pattern does not distinguish isoforms; do not infer isoform-specific staining from location alone. |
| Strength of IHC evidence | HPA marks antibody HPA017578 Approved for IHC, while its tissue profile notes medium staining-to-RNA consistency and pending external verification (HPA: antibody validation; tissue IHC). Use its pattern as an expectation, and retain controls when interpreting discordant samples. |
| IF/ICC Q: Where should signal appear? | A: HPA reports enhanced cytosolic localization and Enhanced ICC validation for HPA017578 (HPA: subcellular ICC-IF; antibody validation). This is an IF/ICC interpretation cue; the paraffin-section IHC pattern remains the primary guide here (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| A known High tissue gives no usable signal. | The run or detection may have failed; HPA reports High staining in colon glandular cells (HPA: colon IHC). | Review the positive control, reagent sequence, antibody dilution, retrieval conditions and detection development (general IHC practice). Change one variable at a time; the payload provides no ABCF1-specific retrieval setting. |
| Staining is weak in a tissue chosen as a positive control. | Cell choice may be the issue: HPA lists hepatocytes and alveolar cells as Low, versus High colon glandular cells (HPA: tissue IHC). | Compare the same cell type and compartment across sections, then check run controls (general IHC practice). Use an HPA High cell population when judging whether the assay can reveal the expected pattern (HPA: tissue IHC). |
| Brown signal outlines cells but cytoplasm stays pale. | That distribution differs from HPA's cytoplasmic tissue profile and is not explained by a transmembrane segment (HPA: tissue IHC; UniProt Q8NE71: topology). | Inspect an independent positive section and reagent controls for the same edge pattern (general IHC practice). Score only convincing intracellular staining after reviewing cell boundaries and counterstain (general IHC practice). |
| Adipocytes stain strongly. | HPA reports adipocytes as Not detected, making nonspecific antibody binding or detection background possibilities (HPA: adipose tissue IHC; general IHC practice). | Check a reagent control and compare staining in an HPA High cell type on a parallel section (HPA: tissue IHC; general IHC practice). Avoid treating adipocyte color alone as evidence of ABCF1 expression. |
| Diffuse color obscures nuclei and cytoplasmic borders. | Excess detection signal or background may prevent a compartment call (general IHC practice). | Inspect controls and review blocking, washes, and development time (general IHC practice). Repeat with conditions that preserve cell detail before scoring the cytoplasmic pattern reported by HPA (HPA: tissue IHC). |
| Only nuclei stain in an otherwise clean section. | UniProt lists nucleoplasm and nuclear envelope, while HPA's tissue IHC summary emphasizes cytoplasm (UniProt Q8NE71: localization; HPA: tissue IHC). | Record the nuclear pattern separately and check positive-tissue and reagent controls (general IHC practice). Do not automatically label it artefact or treat it as confirmation of the HPA cytoplasmic IHC pattern. |
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: its glandular cells should stain (HPA: High in colon glandular cells; M05358 tissue-IHC caption: paraffin-embedded human colon). Run adipose tissue as the negative: adipocytes are listed as not detected (HPA: Not detected in adipocytes); on the colon slide, use non-glandular cells as an internal background reference only where they are visibly unstained, without assuming they are ABCF1-negative (HPA: colon row identifies glandular cells only).
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, subclass and concentration; and, where available, ABCF1-knockout material or a validated immunizing-peptide block (standard IHC practice). For colon chromogenic IHC, block endogenous peroxidase and check for background in blood or inflammatory cells; block endogenous biotin if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: Paraffin-section colon IHC is documented, but the selected SKU caption does not report its fixative (M05358 tissue-IHC caption); no target-specific fixation window or retrieval dependence is reported in the supplied evidence. These data do not establish whether frozen sections or IF are easier, although cytosolic ICC-IF localization is documented (HPA subcellular: Cytosol, enhanced). In colon sections, assess glandular luminal material and endogenous peroxidase as possible sources of chromogenic background (standard IHC practice).
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
These questions focus on chromogenic IHC of paraffin sections, with one IF/ICC entry for the separate guide (selected product caption; HPA subcellular).
What retrieval conditions should I try first for ABCF1 in paraffin sections?
Start with Tris-EDTA pH 9.0 heat-induced antigen retrieval at 95–98 °C for 20 min (page retrieval setting). Let sections cool in the retrieval buffer before washing, and keep heating conditions consistent across the staining run (standard IHC practice). The selected product caption shows ABCF1 staining in paraffin-embedded human colon, but reports no fixative or retrieval method, so it cannot establish which conditions produced that image (selected product caption, M05358). If staining is weak, compare a citrate pH 6.0 fallback on adjacent sections while holding antibody and detection conditions constant (standard IHC practice). Judge recovery by glandular-cell staining and background together (HPA tissue IHC; standard IHC practice).
How should I assess whether fixation is suppressing ABCF1 staining?
ABCF1-specific fixation sensitivity is unknown from the supplied evidence; the selected paraffin-colon caption does not state a fixative (selected product caption, M05358). Record the fixative, fixation duration, tissue thickness, and processing history for each block before comparing staining (standard IHC practice). Use matched sections from blocks with documented handling and run them with the same Tris-EDTA pH 9.0, 95–98 °C, 20 min retrieval setting (page retrieval setting; standard IHC practice). Compare glandular cells in colon against adjacent morphology and background, since these cells show high ABCF1 staining in the HPA tissue profile (HPA tissue IHC). Treat a fixation effect as a hypothesis until matched material separates it from retrieval or detection effects (standard IHC practice).
Should ABCF1 staining be cytoplasmic, nuclear, or membranous?
Expect predominantly cytoplasmic staining in tissue sections: the HPA tissue profile describes cytoplasmic expression in most tissues (HPA tissue IHC). UniProt also lists the nucleoplasm and nuclear envelope, so a nuclear component can be assessed, but should be recorded separately from cytoplasmic signal (UniProt Q8NE71 subcellular; standard IHC practice). ABCF1 has no transmembrane segment, making a crisp plasma-membrane-only pattern difficult to reconcile with its recorded topology (UniProt Q8NE71 topology). In colon, examine glandular cells against an appropriate counterstain and preserve cell boundaries when judging compartmental staining (HPA tissue IHC; standard IHC practice). Compare any unexpected nuclear or rim pattern with morphology, adjacent sections, and antibody-omission controls before assigning it to ABCF1 (standard IHC practice).
Can this IHC antibody distinguish ABCF1 isoforms or modified epitopes?
The record lists 2 ABCF1 isoforms, but the supplied caption gives no antibody epitope or isoform-specific staining evidence (UniProt Q8NE71 isoforms; selected product caption, M05358). Do not label a stained cell as isoform 2 solely from chromogenic intensity or compartment (standard IHC interpretation). Isoform 2 associates with ribosomes through both ABC transporter domains and interacts with EIF2S1 through its N-terminus, but those functions do not identify the catalog antibody's binding site (UniProt Q8NE71 subunit). The record also lists phosphorylation sites, including serines 22 and 24; staining alone cannot establish phosphorylation without a validated modification-specific reagent (UniProt Q8NE71 modified residues; standard IHC practice). Seek documented epitope mapping and an appropriate specificity control before making isoform or modification claims (standard IHC practice).
How can IF help check ABCF1 localisation alongside a cell-type marker?
For multiplex IF/ICC, pair ABCF1 with a marker identifying the expected glandular cells when examining colon, and score colocalisation within individual cells (HPA tissue IHC; standard IF practice). Choose a fluorophore channel away from the specimen's strongest autofluorescence and collect single-label controls before interpreting overlap (standard IF practice). Permeabilise sufficiently to expose intracellular epitopes, because ABCF1 is recorded in the cytosol and nucleoplasm and has no transmembrane segment (UniProt Q8NE71 subcellular and topology; standard IF practice). Optimise permeabilisation against preserved morphology rather than assuming an epitope location that has not been supplied (standard IF practice). HPA reports enhanced cytosolic localisation from ICC/IF, including U2OS images with siRNA and scrambled conditions, but supplies no fixation recipe here (HPA subcellular).
How do I reduce diffuse or granular ABCF1 staining in chromogenic IHC?
First compare the stained section with a primary-antibody omission control and a matched isotype control where appropriate (standard IHC practice). Block endogenous peroxidase before enzyme-based detection, and inspect pigment or precipitate under brightfield because these can mimic chromogenic signal (standard IHC practice). Use a protein block and wash consistently, then titrate the catalog antibody and detection reagents within their documented instructions; no dilution is supplied in this payload (standard IHC practice). Because HPA describes cytoplasmic expression in most tissues, diffuse cellular signal alone is not proof of background (HPA tissue IHC). A useful comparison is strong glandular-cell staining in colon against the reported absence of staining in adipocytes, interpreted with tissue quality and controls (HPA tissue IHC; standard IHC practice).
What is a defensible way to quantify ABCF1 across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the compartment and cell population before scoring; HPA reports high ABCF1 staining in colon glandular cells and cytoplasmic expression in most tissues (HPA tissue IHC). For chromogenic IHC, record the percentage of positive target cells and staining intensity, then calculate an H-score if the study uses prespecified intensity bins (standard IHC practice). Alternatively, report positive-cell density per mm² of viable annotated tissue when cell counts are the outcome (standard IHC practice). Normalise across comparable tissue area, cell type, section quality, staining batch, and exposure to detection reagents (standard IHC practice). Exclude folds, necrosis, and section edges by a prespecified rule, and keep nuclear and cytoplasmic scores separate if both are measured (standard IHC practice).
Which staining patterns would make an ABCF1-positive result questionable?
A convincing colon result shows signal in intact glandular cells with a credible intracellular distribution; HPA reports high glandular-cell staining and predominantly cytoplasmic tissue expression (HPA tissue IHC). A plasma-membrane-only outline is questionable because ABCF1 has no transmembrane segment, while nuclear signal needs separate assessment because UniProt also lists nucleoplasm and nuclear envelope (UniProt Q8NE71 topology and subcellular). Treat staining confined to necrotic regions or section edges as suspect and check adjacent well-preserved tissue (standard IHC practice). Compare signal with antibody-omission and endogenous-peroxidase controls before attributing brown deposits to ABCF1 (standard IHC practice). HPA rates its tissue staining Approved but describes medium consistency with RNA data and pending external verification, so use independent specificity evidence for strong biological claims (HPA tissue IHC).
Boster reagents
Best ABCF1 / ATP-binding cassette sub-family F member 1 IHC Antibodies
The catalog antibody has real IHC data from paraffin-embedded human colon (M05358 IHC image caption); IF/ICC is listed, but no IF image is supplied (M05358 applications; catalog IF image alts).
Real IHC data
Anti-ABCF1 Rabbit Monoclonal Antibody
Cat # M05358
M05358 has an IHC image from paraffin-embedded human colon (M05358 IHC image caption). IHC and IF/ICC are listed applications, with Human and Rat listed as reactive species (M05358 applications; M05358 reactivity).
Which to pick: For tissue IHC, choose M05358: its own image documents staining in paraffin-embedded human colon, while the fixative is unreported (M05358 IHC image caption). For IF/ICC, M05358 is listed for both applications, although no IF image is supplied (M05358 applications; catalog IF image alts). For rat samples, M05358 lists Rat reactivity; it is a rabbit monoclonal antibody, but its supplied IHC image shows human colon only (M05358 reactivity; M05358 host and clone; M05358 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 tissue staining (HPA tissue IHC); nuclear locations annotated (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: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image M05358); Caveat: Catalog text also describes phospho-GluR1; verify specificity (datasheet); Isoform / epitope: 2 isoforms; no TM segment; epitope coverage unknown (UniProt); Regulation: Ubiquitous expression; no stated inducer (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.