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ABCB6 / ATP-binding cassette sub-family B member 6 · IHC design guide
Design Immunohistochemistry for ABCB6
Plan ABCB6 paraffin-section IHC with a starting antibody concentration of 0.5–1 μg/ml (datasheet: PA1723 IHC-P). Epididymal or gallbladder glandular cells provide high-staining reference tissue (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ABCB6 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 | Granular cytoplasm (HPA tissue IHC); organelle membranes (UniProt) | |
| Staining pattern | Granular cytoplasmic staining in most tissues (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Epididymis+4 more · see all | |
| Negative control | Lymph node+2 more · see all |
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 PA1723) | |
| Caveat | ER-to-endolysosomal trafficking may shift the pattern (UniProt) | |
| Regulation | No expression regulator specified (UniProt) | |
| Isoform / epitope | 2 isoforms; check epitope against isoform and topology (UniProt) |
Section 1
Recommended ABCB6 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by one published ABCB6 skin IHC protocol (PMC3818219).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet PA1723); 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-ABCB6, 0.5-1μg/ml (datasheet PA1723) |
| 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 | ABCB6-positive staining in glandular cells of epididymis (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression with a granular pattern in most tissues. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval); the published skin protocol used EDTA pH 8.0 (PMC3818219).
Section 2
What Is the Expected ABCB6 Staining Pattern?
In paraffin-section IHC, expect predominantly granular cytoplasmic ABCB6 staining in many tissues (HPA: tissue IHC profile), with stronger staining in epididymal and gallbladder glandular cells and late spermatids (HPA: High). ABCB6 spans membranes 11 times and occupies several cellular membranes (UniProt Q9NP58 topology and subcellular location). HPA rates its tissue IHC evidence Approved, with medium consistency between staining and RNA expression (HPA: tissue IHC reliability).
| Granular cytoplasmic staining in epididymal or gallbladder glandular cells, or in elongated or late spermatids. | This matches the reported high-staining cell populations (HPA: High in each population). Compare cells within the section: the expected pattern is granular cytoplasm across most tissues, while intensity depends on the sampled cell type (HPA: tissue IHC profile and levels). |
| Predominantly nuclear staining, with little granular cytoplasmic signal, in a paraffin section. | Treat this as a compartment mismatch requiring validation, rather than confirmed ABCB6 IHC: HPA describes granular cytoplasmic tissue staining (HPA: tissue IHC profile). Nucleoplasmic localisation is an ICC-IF observation and does not establish a predominantly nuclear IHC pattern (HPA: subcellular, nucleoplasm approved). |
| Strong stain in germinal center cells, parathyroid glandular cells, or chondrocytes. | Investigate cross-reactivity or endogenous chromogenic activity before assigning target signal (general IHC practice). These specified cell populations were reported as not detected; that rating does not make every cell in their surrounding tissues a negative control (HPA: tissue IHC, Not detected). |
| Uniform color across many compartments, including spaces without cells. | A diffuse field does not resemble the reported granular cytoplasmic pattern (HPA: tissue IHC profile). Consider nonspecific reagent binding, incomplete blocking, or residual chromogen as general IHC causes; inspect a control omitting primary antibody before interpreting cellular staining (general IHC practice). |
| No stain in epididymal or gallbladder glandular cells, or in late spermatids. | These are reported high-staining populations, so an absent result warrants an assay check (HPA: High in these populations). Review tissue preservation, retrieval conditions, primary-antibody performance, and detection controls as general IHC checks; one failed section alone cannot establish absent ABCB6 expression. |
💡Expected ABCB6 appearanceCall a paraffin-section result positive when the relevant cells show discernible granular cytoplasmic staining, strongest in HPA high populations; broad diffuse color or predominantly nuclear stain needs investigation (HPA: tissue IHC profile and levels; general IHC practice).
| Membrane topology and compartment | ABCB6 has 11 transmembrane segments and cytoplasmic and lumenal regions (UniProt Q9NP58 topology). An antibody's epitope and assay conditions govern access in general IHC practice; epitope location is not supplied here. |
| Intracellular distribution | ABCB6 is reported at plasma, Golgi, mitochondrial, and endolysosomal membranes (UniProt Q9NP58 subcellular location). These locations can contribute to cellular staining, but HPA's tissue IHC summary remains granular cytoplasm (HPA: tissue IHC profile). |
| Antibody evidence | HPA046723 is IHC Approved; HPA058011 has no IHC status in the supplied record (HPA: antibodies). Neither entry establishes an IHC Enhanced result (HPA: antibodies). Interpret an unverified antibody's pattern against tissue controls. |
| Isoforms and glycosylation | UniProt lists 2 isoforms and one glycosylation site at residue 6 (UniProt Q9NP58). Their effects on the IHC epitope or staining intensity cannot be inferred from these entries. |
| Endogenous detection activity | If using peroxidase-based chromogenic detection, endogenous enzyme activity can mimic antibody staining (general IHC practice). An appropriate enzyme-blocking step and a control without primary antibody help identify that source. |
| IF/ICC Q: should its localisation dictate the IHC readout? | A: No. Golgi is the supported main ICC-IF location; plasma membrane is also supported, while nucleoplasm and mitochondria are approved additional locations (HPA: subcellular). Score paraffin IHC against its granular cytoplasmic tissue profile (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| A high-staining cell population has no chromogenic signal. | Assay failure is possible because epididymal and gallbladder glandular cells and late spermatids are reported High (HPA: tissue IHC). | Check tissue morphology and a working positive section; then review retrieval, primary-antibody incubation, and detection reagents as general IHC checks. Avoid calling the target absent from this result alone. |
| The whole section looks diffusely stained. | Uniform color conflicts with the granular cytoplasmic tissue profile (HPA: tissue IHC). Nonspecific binding or chromogen background are possible (general IHC practice). | Examine a no-primary control and review blocking, washes, detection-reagent concentration, and chromogen development (general IHC practice). Reassess only cell-associated staining. |
| Nuclei dominate the paraffin-section signal. | The IHC profile is granular cytoplasmic (HPA: tissue IHC); nucleoplasm is an additional ICC-IF localisation (HPA: subcellular, approved). | Compare the slide with a reported high-staining cell population and a no-primary control; verify antibody identity and scoring compartment (general IHC practice). Do not transfer the ICC-IF location directly to IHC. |
| Cells rated Not detected show strong color. | Germinal center cells, parathyroid glandular cells, and chondrocytes were individually rated Not detected (HPA: tissue IHC); nonspecific or endogenous signal remains possible (general IHC practice). | Confirm the cell identity, examine no-primary and detection controls, and compare the pattern with reported high-staining cells (general IHC practice). Evaluate the named cells rather than scoring the whole tissue negative. |
| Only faint staining appears in a low-staining population. | Respiratory epithelial cells in bronchus and macrophages in lung are listed Low (HPA: tissue IHC); faint cellular signal may fit that rating. | Compare morphology and granular cytoplasmic localisation with a high-staining control (HPA: tissue IHC profile and levels). Avoid increasing chromogen development solely to make a Low population appear High (general IHC practice). |
| A membrane outline is seen without clear cytoplasmic granules. | Plasma membrane localisation is reported (UniProt Q9NP58; HPA: subcellular, supported), but tissue IHC is summarized as granular cytoplasm (HPA: tissue IHC). | Assess whether the outline belongs to the expected cell population, then compare a high-staining control and a no-primary control (HPA: tissue IHC; general IHC practice). Record the deviation before calling it target-specific. |
Experiment evidence
IHC & IF images showing ABCB6 staining
PA1723 · IHCAnti-ABCB6 antibody, PA1723, IHC(P) IHC(P): Human Mammary Cancer Tissue
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000115657-ABCB6/tissue/cerebral+cortex#imid_12837723
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000115657-ABCB6/tissue/colon#imid_12837641
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000115657-ABCB6/tissue/liver#imid_12837658
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000115657-ABCB6/tissue/kidney#imid_12837651
HPA · ICC-IF Hep-G2Human Protein Atlas immunofluorescence, Hep-G2, antibody HPA046723; green = ABCB6, blue = nucleus, red = microtubules.
HPA · ICC-IF Hep-G2Human Protein Atlas immunofluorescence, Hep-G2, antibody HPA046723; green = ABCB6, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ABCB6 IHC & IF
🧪Run epididymis first and score its glandular cells for staining (HPA: High in epididymis glandular cells). Run lymph node germinal center cells as the tissue negative (HPA: Not detected); on the epididymis slide, use nonglandular cells to assess background without assuming they lack ABCB6 (HPA: only glandular cells are specified).
Positive control tissue: Epididymis (Glandular cells, HPA High)
Negative control tissue: Lymph node (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ABCB6 in Hep-G2, U-251MG, U2OS, Sperm, with annotated localisation: Golgi apparatus (supported) (HPA subcellular).
Technical controls: Include no-primary and secondary-only controls, an isotype control matched to the primary antibody’s host species and clonality, and ABCB6-knockout material as a biological negative (standard IHC practice). For chromogenic detection, block endogenous peroxidase and check glandular background in the secondary-only control (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and retrieval dependence are unreported in the supplied evidence; the selected PA1723 IHC(P) caption also leaves the fixative unreported (catalog caption: fixative not stated). These data do not establish whether frozen sections or IF/ICC would be easier, although ICC-IF images are available (HPA: subcellular ICC-IF images). Assess glandular background and endogenous peroxidase as potential interpretation artifacts in the positive slide (standard IHC practice).
HPA tissue IHC evidence for ABCB6
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Testis | Elongated or late spermatids | High | Protein (IHC) | HPA → |
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced ABCB6 IHC Tips
Troubleshoot ABCB6 staining in paraffin section chromogenic IHC, with one companion question on multiplex immunofluorescence.
How should I retrieve ABCB6 when paraffin section staining is weak?
Start with citrate buffer at pH 6.0 and heat induced epitope retrieval at 95–98 °C for 20 minutes (page retrieval rule). Compare a retrieved section with an untreated section while keeping antibody concentration, detection, and development time constant, so a change in signal can be attributed to retrieval. Score intracellular granular staining in glandular cells alongside background in germinal center cells (HPA tissue IHC: high in gallbladder glandular cells; not detected in lymph node germinal center cells; granular cytoplasmic profile). If signal remains weak, adjust heating time in small increments on matched sections and reject conditions that spread staining across tissue compartments.
Could fixation explain weak or diffuse ABCB6 staining in paraffin sections?
ABCB6 specific fixation sensitivity is unknown from the supplied evidence; the selected IHC(P) caption identifies human mammary cancer tissue but does not state a fixative (PA1723 tissue IHC caption). Record the actual fixative and fixation duration for each specimen, then compare sections processed together before assigning weak staining to antigen loss. If fixation histories differ, standardize section thickness, citrate retrieval at pH 6.0 for 20 minutes, and chromogen development before comparing intensity (page retrieval rule; standard IHC practice). Assess cellular morphology and background alongside staining; neither a granular tissue pattern nor ABCB6 membrane topology establishes a fixation effect (HPA tissue IHC; UniProt Q9NP58 topology).
Which staining compartments are plausible for ABCB6 in chromogenic tissue IHC?
Expect a predominantly granular cytoplasmic pattern in tissue sections (HPA tissue IHC: granular cytoplasmic expression in most tissues). ABCB6 is assigned to Golgi, cell, mitochondrial outer, endosomal, and lysosomal membranes, so a membrane associated or punctate intracellular pattern can be biologically plausible (UniProt Q9NP58 subcellular location). HPA cell imaging supports Golgi as the main location and also supports plasma membrane staining, but those cell observations do not set a required pattern for every tissue (HPA subcellular). Compare the stain with cell outlines and organelle marker controls on matched sections; diffuse nuclear only chromogen should prompt a review of background and detection conditions.
How can epitope position change the apparent ABCB6 IHC pattern?
Map the antibody immunogen to the 842 amino acid sequence before interpreting a negative section; ABCB6 has 2 listed isoforms and 11 transmembrane segments (UniProt Q9NP58 sequence, isoforms, topology). The 551–842 region is cytoplasmic, whereas several short loops are lumenal, so epitope position can affect accessibility after section processing (UniProt Q9NP58 topology; standard IHC practice). A glycosylation site is annotated at residue 6, but its effect on this antibody's staining is unreported (UniProt Q9NP58 glycosylation). When an expected cell population stains weakly, confirm the immunogen sequence and isoform coverage, then compare retrieval conditions without treating absent staining as proof of absent protein.
How should I assess ABCB6 localisation in a companion multiplex IF experiment?
Use a cell type marker in a separate channel to identify the population under study, such as glandular cells when examining gallbladder sections (HPA tissue IHC: high in gallbladder glandular cells). Select fluorophores after imaging unstained tissue for autofluorescence, and use spectral separation and single stain controls to check bleed through (standard IF practice). Plan permeabilisation around the antibody's documented epitope: cytoplasmic regions such as 551–842 require intracellular access, while lumenal loops may require access to internal membranes (UniProt Q9NP58 topology). Compare puncta with an appropriate organelle marker, and do not assume the PA1723 tissue IHC caption validates IF conditions (PA1723 tissue IHC caption).
How can I reduce diffuse brown background without losing ABCB6 signal?
First compare a no primary control with the stained section to locate background from the detection system (standard chromogenic IHC practice). Quench endogenous peroxidase before chromogen development, block nonspecific binding, and shorten overly strong DAB development while keeping matched sections under identical conditions (standard chromogenic IHC practice). Preserve the expected granular cytoplasmic readout during optimization (HPA tissue IHC: granular cytoplasmic profile), and monitor a lower staining reference such as lymph node germinal center cells (HPA tissue IHC: not detected). Record retrieval at pH 6.0 for 20 minutes and compare one variable at a time; excessive retrieval can complicate interpretation by degrading morphology (page retrieval rule; standard IHC practice).
What should I score when comparing ABCB6 across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, since ABCB6 shows granular cytoplasmic staining across many tissues (HPA tissue IHC: granular cytoplasmic profile). For each predefined region, report the percentage of positive cells and an H score from intensity categories 0–3, or count positive cells per mm² when cell density is the outcome (standard IHC scoring practice). Normalize to the number of eligible cells or analyzed tissue area, and keep section processing, DAB development, and image thresholds consistent. Report scores by cell type rather than whole section averages when populations differ; gallbladder glandular cells and lymph node germinal center cells have different reference staining levels (HPA tissue IHC: high; not detected).
When is apparent ABCB6 positivity more likely to be artefact?
Treat staining as more convincing when it is cellular and granular in an expected population, such as gallbladder glandular cells (HPA tissue IHC: granular cytoplasmic profile; high in gallbladder glandular cells). Strong isolated nuclear staining warrants scrutiny because HPA tissue IHC describes a cytoplasmic pattern, although cell imaging also lists nucleoplasm as an additional location (HPA tissue IHC; HPA subcellular: nucleoplasm approved). Examine section edges and necrotic areas for uneven deposit, and compare a no primary control to identify endogenous enzyme or detection background (standard chromogenic IHC practice). Avoid declaring every unstained cell negative: lymph node germinal center cells are a useful low signal reference, and tissue expression varies (HPA tissue IHC: not detected in germinal center cells; low tissue specificity).
Boster reagents
Best ABCB6 / ATP-binding cassette sub-family B member 6 IHC Antibodies
The IHC-validated anti-ABCB6 antibody has an IHC(P) image of human mammary cancer tissue (PA1723 image caption). Human ICC is listed, but no IF image is supplied (PA1723 applications/reactivity; IF image alts).
Real IHC data
Anti-ABCB6 Antibody ®
Cat # PA1723
PA1723 shows IHC(P) staining of human mammary cancer tissue (PA1723 image caption). It is listed for human IHC and ICC, with no IF image supplied (PA1723 applications/reactivity; IF image alts).
Which to pick: Choose PA1723 for human paraffin-section IHC: its own image shows human mammary cancer tissue, and the fixative is unreported (PA1723 image caption). For human ICC, PA1723 is listed at 0.5–1 μg/ml; no IF image is supplied (PA1723 applications/IF dilution/IF image alts). There is no supported cross-species choice because PA1723 lists human reactivity only (PA1723 reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9NP58 (ABCB6_HUMAN, ATP-binding cassette sub-family B member 6).
- Human Protein Atlas. ABCB6 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. ABCB6 subcellular location (ICC-IF): Mainly localized to the Golgi apparatus. In addition localized to the nucleoplasm, plasma membrane, cytosol, mitochondria, mid piece and principal piece..
- Human Protein Atlas. ABCB6 antibody validation summary (2 antibodies).
- Understanding the role of iron/heme metabolism in the anti‑inflammatory effects of natural sulfur molecules against lipopolysaccharide‑induced inflammation. Molecular medicine reports 2025 — PMC12046942.
- Novel mutations of ABCB6 associated with autosomal dominant dyschromatosis universalis hereditaria. PloS one 2013 — PMC3818219.
- Protoporphyrin IX (PpIX) Fluorescence during Meningioma Surgery: Correlations with Histological Findings and Expression of Heme Pathway Molecules. Cancers 2023 — PMC9818642.
- Elevated Glutathione Peroxidase 2 Expression Promotes Cisplatin Resistance in Lung Adenocarcinoma. Oxidative medicine and cellular longevity 2020 — PMC7079220.
- PubMed PMID:10837493 — UniProt-cited evidence.
- PubMed PMID:11955620 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ABCB6 (ATP-binding cassette sub-family B member 6). Expected localisation Granular cytoplasm (HPA tissue IHC); organelle membranes (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: PA1723 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Epididymis (Glandular cells); negative Lymph node. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet PA1723); 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-ABCB6, 0.5-1μg/ml (datasheet PA1723); 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 PA1723); Caveat: ER-to-endolysosomal trafficking may shift the pattern (UniProt); Isoform / epitope: 2 isoforms; check epitope against isoform and topology (UniProt); Regulation: No expression regulator specified (UniProt). Evidence: UniProt Q9NP58, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the Golgi apparatus. In addition localized to…, Datasheet PA1723, PMC3818219. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ABCB6/immunohistochemistry-ihc-guide.