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ABCF2 / ATP-binding cassette sub-family F member 2 · IHC design guide
Design Immunohistochemistry for ABCF2
Plan ABCF2 paraffin IHC around the cytoplasmic staining reported in most tissues (HPA tissue IHC). Compare stained sections with appropriate controls and start with the catalog antibody’s 1:50–1:200 IHC dilution range (datasheet).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ABCF2 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 staining in most tissues (HPA tissue IHC) | |
| Staining pattern | Colon glandular cells: medium, cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue+4 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 | Staining may reflect proteins from more than one gene (HPA tissue IHC) | |
| Regulation | Expression regulation unreported (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope effects undetermined (UniProt) |
Section 1
Recommended ABCF2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet) is followed by published ABCF2 staining protocols from uterine cancer and hepatocellular carcinoma studies (PMC2584961; PMC6998385).
| Sample | Paraffin-embedded mouse lung tissue; fixative not specified (datasheet A10873-1) |
| Fixation | Image fixative and duration unreported (datasheet A10873-1); 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-ABCF2, 1:50-1:200 (datasheet A10873-1) |
| 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 | ABCF2-positive staining in glandular cells of adrenal gland (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page antigen retrieval); the hepatocellular carcinoma study used citric acid at 95 °C for 20 min (PMC6998385).
Section 2
What Is the Expected ABCF2 Staining Pattern?
In paraffin-section IHC, expect mainly cytoplasmic ABCF2 staining across many tissues, including glandular, neuronal and squamous epithelial cells (HPA tissue IHC: cytoplasmic expression in most tissues; listed cell types). UniProt reports no transmembrane segment and gives no subcellular annotation (UniProt Q9UG63 topology). Interpret the pattern cautiously: HPA rates tissue IHC Approved, but reports medium consistency with RNA data and warns that the antibodies target proteins from more than one gene (HPA tissue IHC).
| Cytoplasmic staining in colon or duodenal glandular cells, or cortical neurons, at moderate intensity. | Consistent with HPA's Medium staining in those cell types (HPA tissue IHC). Compare cells within the section; a positive image alone cannot resolve HPA's multiple-gene targeting caution (HPA tissue IHC). |
| Predominantly nuclear or sharply membrane-restricted staining, with little cytoplasmic signal. | Departing from HPA's cytoplasmic tissue profile warrants an artefact or specificity check (HPA tissue IHC). UniProt lists no transmembrane segment, but topology alone cannot rule out every form of membrane association (UniProt Q9UG63 topology). |
| Strong signal in adipocytes or glomerular cells while expected positive cells are weak. | These cells are listed as Not detected, so investigate cross-reactivity or endogenous chromogenic activity with appropriate controls (HPA tissue IHC; general IHC practice). One unexpected cell population does not establish which cause applies. |
| Uniform color over tissue, stroma and nominally negative cells. | A diffuse field is difficult to reconcile with HPA's cell-resolved profile, including Not detected populations (HPA tissue IHC). In general IHC practice, check background from detection reagents, inadequate blocking or incomplete washing before assigning it to ABCF2. |
| No cytoplasmic signal in colon glandular cells or pancreatic exocrine glandular cells. | Both are listed as Medium, so a blank result needs investigation (HPA tissue IHC). Check section quality, retrieval and the IHC-validated antibody's prescribed detection workflow (general IHC practice); HPA supplies no ABCF2-specific fixation-sensitivity claim. |
💡Expected ABCF2 appearanceCall positive when cell-resolved cytoplasmic color is visible at roughly Medium intensity in HPA-listed glandular or neuronal cells; dominant nuclear, membrane-only or uniformly diffuse color needs control-based review (HPA tissue IHC).
| Tissue and cell selection | HPA reports low tissue specificity and cytoplasmic expression in most tissues; its Medium and Not detected labels apply to the named cell populations, not every cell in an organ (HPA tissue IHC). |
| Topology and processing | UniProt lists no transmembrane segment, signal peptide or propeptide, and one chain spanning residues 1–623 (UniProt Q9UG63). These facts support cautious compartment interpretation; they do not predict retrieval needs. |
| Isoforms and epitope coverage | UniProt lists two isoforms and two ABC transporter domains, at residues 86–325 and 396–613 (UniProt Q9UG63). The supplied record does not map antibody epitopes, so isoform coverage cannot be inferred. |
| Post-translational modifications | UniProt lists modified residues including Thr218, Lys304 and Ser512 (UniProt Q9UG63). Without an epitope map, their effect on staining or retrieval is unknown. |
| Antibody validation | HPA lists HPA020091, HPA021815 and HPA030388 as IHC Approved; its tissue profile warns of multiple-gene targeting and medium RNA concordance (HPA antibodies; HPA tissue IHC). Treat staining as supportive, not uniquely ABCF2-specific. |
| What should IF/ICC show? | HPA describes mainly cytosolic signal, labels the location uncertain, and notes multiple-gene targeting; A-431, U-251MG and U2OS have ICC-IF images (HPA subcellular). Use the separate IF/ICC guide for that application. |
| Situation | Likely cause | Next action |
|---|---|---|
| Colon or duodenum glandular cells show no signal. | Both are HPA Medium populations; the failed step is undetermined (HPA tissue IHC). | Verify the IHC-validated antibody and its stated workflow, then review retrieval, detection and section integrity (general IHC practice). Do not infer ABCF2-specific fixation sensitivity. |
| Signal is predominantly nuclear. | That pattern conflicts with HPA's cytoplasmic profile; the source does not identify its cause (HPA tissue IHC). | Compare a second HPA-listed Medium tissue and controls; review counterstain and chromogen localization before scoring the signal (HPA tissue IHC; general IHC practice). |
| Adipocytes or marrow hematopoietic cells stain strongly. | Both populations are HPA Not detected; cross-reactivity or endogenous detection activity remains possible (HPA tissue IHC; general IHC practice). | Run no-primary and detection controls, inspect cell boundaries, and compare an HPA Medium population in the same run (HPA tissue IHC; general IHC practice). |
| Brown color covers tissue and surrounding background. | Diffuse color obscures HPA's cell-resolved pattern; reagent background is a general IHC possibility (HPA tissue IHC; general IHC practice). | Check no-primary control, blocking, washes and detection development; score only localized cellular signal (general IHC practice). |
| One antibody stains a tissue that another does not. | HPA lists multiple IHC Approved antibodies but cautions that antibodies target proteins from more than one gene (HPA antibodies; HPA tissue IHC). | Compare the same named cell population and controls across antibodies; report discordance rather than assigning it to an ABCF2 isoform without epitope evidence (HPA tissue IHC; UniProt Q9UG63 isoforms). |
| A nominally negative tissue has scattered stained cells. | HPA's Not detected entries name specific cell populations; they do not establish that every other cell in that tissue is negative (HPA tissue IHC). | Identify the stained cell type, compare its HPA entry if supplied, and use controls to assess background before calling the section positive or negative (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing ABCF2 staining
A10873-1 · IHCImmunohistochemistry of paraffin-embedded mouse lung using ABCF2 antibody at dilution of 1:100 (x40 lens).
A10873-1 · IHCImmunohistochemistry of paraffin-embedded human stomach using ABCF2 antibody at dilution of 1:100 (x40 lens).
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000285292-ABCF2/tissue/cerebral+cortex#imid_5936512
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000285292-ABCF2/tissue/colon#imid_5936438
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000285292-ABCF2/tissue/liver#imid_5936439
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000285292-ABCF2/tissue/kidney#imid_5936424
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA020091; green = ABCF2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA020091; green = ABCF2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ABCF2 IHC & IF
🧪Run colon first: glandular cells should stain at the HPA Medium level (HPA: colon glandular cells, Medium). Use cervix glandular cells as the negative tissue (HPA: cervix glandular cells, Not detected); on the colon slide, use unstained nonglandular cells as a background reference, without assuming they are biologically ABCF2-negative (HPA: colon glandular-cell staining only).
Positive control tissue: Adrenal gland (Glandular cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ABCF2 in A-431, U-251MG, U2OS, with annotated localisation: Cytosol (uncertain) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and control-antibody sections: match the primary’s host and isotype for a monoclonal antibody, or use host-matched nonimmune IgG for a polyclonal antibody (standard IHC practice). Where available, use ABCF2 knockout material or an immunizing-peptide block as a biological specificity control, and quench endogenous peroxidase while checking inflammatory cells for residual chromogen signal in colon (standard IHC practice; HPA: colon selected as positive tissue).
⚠️Feasibility: A target-specific fixation window and the effect of antigen retrieval are unreported in the supplied evidence; optimize retrieval on paraffin sections empirically (standard IHC practice). The selected-SKU caption shows paraffin-embedded mouse lung at 1:100, but its fixative is unreported (A10873-1 tissue-IHC caption). The supplied evidence does not establish that frozen sections or IF are easier; for colon IHC, assess inflammatory-cell peroxidase background before scoring glandular staining (HPA: colon glandular cells, Medium; standard IHC practice).
HPA tissue IHC evidence for ABCF2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene.). 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | Medium | Protein (IHC) | HPA → |
| Colon | Glandular cells | Medium | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ABCF2 IHC Tips
Troubleshoot ABCF2 staining in paraffin sections by checking retrieval, compartment, cell type and controls before comparing staining intensity (HPA tissue IHC; HPA subcellular).
How should I adjust retrieval when ABCF2 staining is weak in paraffin sections?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 min (page retrieval specification). After cooling, compare the ABCF2 signal and section integrity with a control section processed in the same run (standard IHC practice). If staining remains weak, test a modest change in heating time on adjacent sections while keeping antibody dilution and detection constant; excessive heating can damage tissue and increase background (standard IHC practice). The selected antibody has a paraffin-section mouse lung image at 1:100, but its caption does not report the fixative or retrieval method, so that image cannot establish an optimal retrieval condition (catalog antibody caption, A10873-1).
Could fixation explain inconsistent ABCF2 staining across my paraffin sections?
ABCF2-specific sensitivity to fixation is unknown from the supplied evidence; the selected mouse lung caption identifies paraffin embedding but does not state a fixative (catalog antibody caption, A10873-1). Record each specimen’s fixative, fixation duration and processing history before comparing chromogenic signal across runs (standard IHC practice). If those variables differ, process adjacent sections together and compare a consistent internal tissue feature or a separately validated control section (standard IHC practice). Do not treat differences in HPA tissue staining as evidence that ABCF2 tolerates one fixation condition better than another; those observations describe expression patterns, not fixation response (HPA tissue IHC).
Where should ABCF2 staining appear, and how should I assess unexpected nuclear signal?
Expect predominantly cytoplasmic staining in tissue sections, while treating that expectation cautiously because the HPA tissue profile reports cytoplasmic expression and warns that its antibody may recognize proteins from more than one gene (HPA tissue IHC). HPA cell imaging places the signal mainly in the cytosol but calls that location uncertain for the same specificity reason (HPA subcellular). ABCF2 has no annotated transmembrane segment, so a crisp plasma-membrane rim needs independent validation before assignment to ABCF2 (UniProt Q9UG63 topology). For strong nuclear staining, compare neighboring cells, a no-primary control and an independent ABCF2 reagent or perturbation where available (standard IHC practice).
Can this antibody distinguish ABCF2 isoforms or a modified epitope in tissue?
ABCF2 has 2 listed isoforms, but the supplied caption does not identify the selected antibody’s epitope or establish isoform specificity (UniProt Q9UG63; catalog antibody caption, A10873-1). Its annotated ABC transporter domains span residues 86–325 and 396–613; examples of modified residues include Thr218, Lys304 and Ser512 (UniProt Q9UG63). Those annotations alone cannot show whether fixation, retrieval or modification changes antibody binding (UniProt Q9UG63; standard IHC practice). If isoform-level interpretation matters, obtain an epitope map and compare independently validated isoform-selective material under the same staining conditions before assigning a chromogenic signal to either isoform (standard IHC practice).
How can I check ABCF2 localisation by multiplex IF alongside the IHC result?
Use IF as a separate validation experiment and multiplex ABCF2 with a marker for the cell population being assessed, such as lung macrophages if comparing with the HPA lung entry (HPA tissue IHC: low in macrophages; standard IF practice). Choose spectrally separated fluorophores after imaging an unstained section, since tissue autofluorescence can overlap dim channels (standard IF practice). For an intracellular ABCF2 epitope, apply a mild permeabilisation step after fixation and compare it with an unpermeabilised condition; ABCF2 has no annotated transmembrane segment, while this antibody’s precise epitope is unreported (UniProt Q9UG63 topology; catalog antibody caption, A10873-1). Include single-color controls for channel bleed-through and assess whether the resulting pattern is predominantly cytosolic (standard IF practice; HPA subcellular: uncertain cytosol).
What should I change when ABCF2 chromogenic staining is diffuse or widespread?
First compare a no-primary section with the stained section to separate detection-system background from antibody-dependent signal (standard IHC practice). Check the peroxidase block before DAB development, then shorten development or reduce primary concentration in a controlled titration if diffuse color persists (standard chromogenic IHC practice). The selected paraffin-section mouse lung image used 1:100; treat that as a reported image condition, not a proven optimum for other specimens (catalog antibody caption, A10873-1). Because HPA reports cytoplasmic expression in most tissues and cautions that its antibody can recognize more than one gene, diffuse staining still needs specificity controls before interpretation (HPA tissue IHC).
How should I score ABCF2 staining when cell populations differ between sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring; HPA reports cytoplasmic tissue expression and lists medium staining in colon glandular cells but low staining in lung macrophages (HPA tissue IHC). For chromogenic sections, report the percentage of positive target cells and an H-score of 0–300 using intensity grades 0–3, with thresholds set from controls (standard IHC scoring practice). If counting discrete positive cells, report density per mm² of evaluable tissue and exclude folds, necrosis and section edges (standard histology scoring practice). Normalize comparisons to the number of eligible cells or evaluable area, and keep staining and imaging settings consistent across groups (standard IHC practice).
How do I distinguish genuine ABCF2 staining from artefact in a positive section?
A credible result should fit the expected cytoplasmic pattern and the cell population being evaluated; HPA reports medium staining in pancreatic exocrine glandular cells and no detection in heart cardiomyocytes (HPA tissue IHC). Treat an isolated membrane rim or strong nuclear signal as a reason to seek corroboration, given ABCF2’s lack of an annotated transmembrane segment and HPA’s uncertain cytosolic assignment (UniProt Q9UG63 topology; HPA subcellular). Exclude edge staining, necrotic areas and color present in a no-primary control, and check peroxidase blocking when endogenous enzyme activity is suspected (standard chromogenic IHC practice). Interpret apparent agreement with HPA cautiously because its tissue and subcellular entries warn of antibody recognition across multiple genes (HPA tissue IHC; HPA subcellular).
Boster reagents
Best ABCF2 / ATP-binding cassette sub-family F member 2 IHC Antibodies
A10873-1 has paraffin-section IHC images from mouse lung and human stomach (catalog image captions). IF is listed, but no IF image is supplied (catalog applications; IF image alts).
Real IHC data
Anti-ABCF2 Antibody
Cat # A10873-1
A10873-1 has IHC images of paraffin-embedded mouse lung and human stomach at 1:100 (catalog image captions). It lists IF and Human, Mouse, and Rat reactivity, with no IF image supplied (catalog applications, reactivity, IF image alts).
Which to pick: For tissue IHC, choose A10873-1: its own captions show paraffin-embedded mouse lung and human stomach at 1:100, within its listed 1:50–1:200 IHC range (catalog image captions; IHC dilution). For IF/ICC, A10873-1 lists IF at 1:50–1:200, but supplies no IF image or ICC validation; its clone is unspecified (catalog applications, IF dilution, IF image alts, clone). For cross-species work, A10873-1 lists Human, Mouse, and Rat reactivity; its IHC captions document mouse and human only, and do not report the fixative (catalog reactivity; image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9UG63 (ABCF2_HUMAN, ATP-binding cassette sub-family F member 2).
- Human Protein Atlas. ABCF2 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. ABCF2 subcellular location (ICC-IF): Mainly localized to the cytosol. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. ABCF2 antibody validation summary (3 antibodies).
- Can ABCF2 protein expression predict the prognosis of uterine cancer?. British journal of cancer 2008 — PMC2584961.
- CircRNA_000864 Upregulates B-cell Translocation Gene 2 Expression and Represses Migration and Invasion in Pancreatic Cancer Cells by Binding to miR-361-3p. Frontiers in oncology 2020 — PMC7793745.
- ABCF2, an Nrf2 target gene, contributes to cisplatin resistance in ovarian cancer cells. Molecular carcinogenesis 2017 — PMC5509336.
- Circ-TCF4.85 silencing inhibits cancer progression through microRNA-486-5p-targeted inhibition of ABCF2 in hepatocellular carcinoma. Molecular oncology 2020 — PMC6998385.
- PubMed PMID:10944468 — UniProt-cited evidence.
- PubMed PMID:17974005 — UniProt-cited evidence.
- PubMed PMID:12853948 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ABCF2 (ATP-binding cassette sub-family F member 2). Expected localisation Cytoplasmic staining in most tissues (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A10873-1 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse lung tissue; fixative not specified (datasheet A10873-1); 2. Fixation: Image fixative and duration unreported (datasheet A10873-1); 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-ABCF2, 1:50-1:200 (datasheet A10873-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining may reflect proteins from more than one gene (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope effects undetermined (UniProt); Regulation: Expression regulation unreported (UniProt). Evidence: UniProt Q9UG63, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the cytosol. Caution: Based on antibodies…, Datasheet A10873-1, PMC2584961, PMC6998385. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ABCF2/immunohistochemistry-ihc-guide.