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ABCC5 / ATP-binding cassette sub-family C member 5 · IHC design guide
Design Immunohistochemistry for ABCC5
Plan ABCC5 paraffin IHC with the catalog antibody at 2–5 μg/mL and chromogenic detection (datasheet A03040-1). Assess cytoplasmic tissue staining (HPA tissue IHC) alongside the expected membrane localization (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ABCC5 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) | |
| Staining pattern | Cytoplasmic staining in kidney tubules and germinal center cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A03040-1) | |
| Positive control | Appendix+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). Selected-image fixative and duration unreported (datasheet A03040-1); verify before use. | |
| Caveat | Staining and RNA levels show low consistency (HPA tissue IHC) | |
| Regulation | Low tissue specificity (HPA tissue IHC) | |
| Isoform / epitope | 5 isoforms; extracellular versus cytoplasmic epitope matters (UniProt) |
Section 1
Recommended ABCC5 IHC & IF Protocols
The catalog antibody has an IHC-P protocol (datasheet A03040-1). The published ABCC5 IHC method below uses OCT-embedded sections (PMC4053136).
| Sample | Paraffin-embedded human gastric cancer tissue; fixative not specified (datasheet A03040-1) |
| Fixation | Image fixative and duration unreported (datasheet A03040-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A03040-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03040-1) |
| Primary antibody | Rabbit anti-ABCC5, 2-5μg/ml (datasheet A03040-1) |
| Primary incubation | Overnight at 4 °C (datasheet A03040-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A03040-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ABCC5-positive staining in lymphoid tissue of appendix (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in several tissues. No signal in the no-primary control. |
💡Decision noteFor paraffin sections, start with heat-mediated EDTA pH 8.0 retrieval (datasheet A03040-1). The published OCT-section method does not specify retrieval (PMC4053136).
Section 2
What Is the Expected ABCC5 Staining Pattern?
ABCC5 is a 13-pass membrane transporter usually routed to the basolateral plasma membrane; brain capillary endothelial cells are an apical exception (UniProt O15440 topology and subcellular location). HPA reports mainly cytoplasmic tissue IHC staining, including medium staining in kidney tubule cells, placental trophoblasts, and lymph node germinal center cells (HPA tissue IHC). Treat these examples as provisional controls because HPA rates its tissue IHC profile Uncertain (HPA tissue IHC).
| Membrane-associated staining in the expected cells, with possible cytoplasmic staining. | This fits UniProt membrane localization and HPA's reported cytoplasmic tissue IHC profile. Assess the two observations separately: HPA's IHC profile has low agreement with RNA expression and is pending retesting, so cytoplasmic staining alone is weak evidence of specificity (UniProt O15440 subcellular location; HPA tissue IHC, Uncertain). |
| Strong staining confined to nuclei, with no convincing membrane-associated pattern. | This is discordant with the principal membrane localization described by UniProt. HPA reports additional nucleoplasmic localization in approved ICC-IF, but that finding does not validate a nuclear-only paraffin IHC result. Check compartment scoring and controls before calling it ABCC5 (UniProt O15440 subcellular location; HPA subcellular ICC-IF). |
| Prominent staining in a cell population reported as not detected by HPA. | Consider antibody cross-reactivity or endogenous chromogen-generating activity, especially if staining also appears in controls (standard IHC practice). HPA's “not detected” calls are assay observations, not proof that those cells cannot express ABCC5; its tissue IHC reliability is Uncertain (HPA tissue IHC). |
| Diffuse color across cells, stroma, or the whole section without a discernible cellular pattern. | This favors nonspecific background over a localizable ABCC5 result. Review the detection and blocking controls, then assess whether staining follows cell boundaries or a reproducible cellular compartment (standard IHC practice; UniProt O15440 subcellular location). |
| No detectable signal in kidney tubule cells or placental trophoblasts. | These are candidate positive reference cells because HPA reports medium staining in each, but its tissue IHC profile is Uncertain. Check the assay's positive control and detection steps before interpreting a blank specimen as absent ABCC5 (HPA tissue IHC; standard IHC practice). |
💡Expected ABCC5 appearanceA plausible positive shows discernible membrane-associated staining, potentially accompanied by cytoplasmic signal, in cells such as kidney tubule cells or placental trophoblasts at roughly medium HPA-reported intensity; widespread featureless color or isolated nuclear staining needs further validation (UniProt O15440 subcellular location; HPA tissue IHC, Uncertain; HPA subcellular ICC-IF).
| Compartment and polarity | UniProt places ABCC5 at basolateral and apical cell membranes, endosome membranes, cytoplasmic granules, and the Golgi lumen. Most cells route it basolaterally; brain capillary endothelial cells are an apical exception. Section orientation and cell boundaries therefore matter when judging a membrane pattern (UniProt O15440 subcellular location; standard IHC practice). |
| Tissue IHC confidence | HPA describes cytoplasmic expression in several tissues but rates the tissue profile Uncertain because antibody staining and RNA expression show low consistency; retesting is pending. Use its medium and not-detected calls as observed patterns, not definitive expression thresholds (HPA tissue IHC). |
| Antibody validation by application | HPA lists HPA052295 tissue IHC as Uncertain and HPA044067 ICC as Approved; these labels apply to different antibodies and applications. HPA's approved plasma membrane, junctional, and additional nucleoplasmic ICC-IF locations cannot establish the catalog antibody's paraffin IHC specificity (HPA antibodies; HPA subcellular ICC-IF). |
| Isoforms and epitope coverage | UniProt lists 5 ABCC5 isoforms. If comparing antibodies, check each disclosed immunogen or epitope against the isoform of interest before treating divergent staining as biological; no epitope map is supplied here, so coverage cannot be predicted (UniProt O15440 isoforms; standard IHC practice). |
| Topology and antigen access | UniProt annotates 13 transmembrane segments and 8 glycosylation sites. These features make epitope position relevant when reviewing an antibody's documentation, but this record supplies no antibody epitope or target-specific retrieval response; choose retrieval conditions from the antibody's IHC instructions and controls (UniProt O15440 topology and glycosylation; standard IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Blank section or weak signal in a candidate positive control. | Detection failure, unsuitable antibody conditions, or a specimen below the assay's detection limit are possibilities; HPA's medium calls are uncertain (HPA tissue IHC; standard IHC practice). | Verify that the IHC-validated antibody and detection controls work, then review the documented retrieval, dilution, and incubation conditions. Compare with kidney tubule or placental trophoblast reference cells without treating either as a guaranteed positive (HPA tissue IHC; standard IHC practice). |
| Predominantly cytoplasmic staining with little visible membrane signal. | HPA reports cytoplasmic tissue IHC staining, while UniProt emphasizes membrane localization; the discrepancy cannot be resolved from these sources alone (HPA tissue IHC; UniProt O15440 subcellular location). | Record cytoplasmic and membrane staining separately, inspect cell boundaries and section morphology, and seek an independent specificity check before assigning the cytoplasmic signal to ABCC5 (standard IHC practice; HPA tissue IHC, Uncertain). |
| Nuclear-only staining in paraffin IHC. | HPA's additional nucleoplasmic finding comes from approved ICC-IF, whereas UniProt lists principally membrane and intracellular membrane-associated locations (HPA subcellular ICC-IF; UniProt O15440 subcellular location). | Check negative detection controls and compare the same antibody's staining across expected cell types; report nuclear-only IHC as unresolved unless independently validated (standard IHC practice; HPA tissue IHC, Uncertain). |
| Color appears in cells HPA calls not detected. | Cross-reactivity or endogenous detection activity may contribute, although an HPA not-detected result does not exclude genuine expression (standard IHC practice; HPA tissue IHC, Uncertain). | Inspect no-primary and detection-system controls, assess whether the signal matches cell morphology, and avoid using one HPA not-detected cell type as a definitive specificity test (standard IHC practice; HPA tissue IHC). |
| Diffuse background obscures cellular localization. | Nonspecific antibody or detection-system background can make a membrane-associated pattern unreadable (standard IHC practice; UniProt O15440 subcellular location). | Check blocking, washes, antibody dilution, and detection controls according to the assay instructions; score ABCC5 only where a reproducible cellular pattern remains interpretable (standard IHC practice). |
| IHC and ICC-IF show different compartments. | HPA's tissue IHC profile is Uncertain, while its plasma membrane, cell junction, and additional nucleoplasmic ICC-IF locations are Approved; antibody identity and application differ in the supplied records (HPA antibodies; HPA tissue IHC; HPA subcellular ICC-IF). | Document the antibody and application used for each result. Compare compartment calls within each assay's controls and do not transfer ICC-IF approval to paraffin IHC (HPA antibodies; standard IHC practice). |
Experiment evidence
IHC & IF images showing ABCC5 staining
A03040-1 · IHCIHC analysis of MRP5/ABCC5 using anti-MRP5/ABCC5 antibody (A03040-1). MRP5/ABCC5 was detected in paraffin-embedded section of human gastric
A03040-1 · IHCIHC analysis of MRP5/ABCC5 using anti-MRP5/ABCC5 antibody (A03040-1). MRP5/ABCC5 was detected in paraffin-embedded section of human mammary
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000114770-ABCC5/tissue/cerebral+cortex#imid_14737564
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000114770-ABCC5/tissue/colon#imid_14736850
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000114770-ABCC5/tissue/liver#imid_14736835
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000114770-ABCC5/tissue/kidney#imid_14736836
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA044067; green = ABCC5, blue = nucleus, red = microtubules.
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA044067; green = ABCC5, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ABCC5 IHC & IF
🧪Run kidney first: tubular cells should stain at the Medium level (HPA: Kidney, cells in tubules, Medium). Use adipose tissue as a negative comparator (HPA: Adipose tissue, adipocytes, Not detected); on the kidney slide, cells outside the expected tubular pattern should lack specific staining, with their status checked empirically.
Positive control tissue: Appendix (Lymphoid tissue, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ABCC5 in MCF-7, RT-4, with annotated localisation: Plasma membrane (approved), Cell Junctions (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a rabbit IgG isotype control matched to the primary antibody’s clonality if known, and an ABCC5-knockout biological negative (datasheet: rabbit primary; standard IHC practice). For kidney sections, block endogenous peroxidase and check for endogenous biotin before using the caption’s biotin-based detection system (caption: biotinylated secondary and SABC; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A03040-1 tissue-IHC caption does not state a fixative (caption: fixative not stated). The demonstrated paraffin-section workflow uses heat-mediated retrieval in EDTA, pH 8.0; this supports starting there but does not establish that retrieval is required for every specimen (caption: EDTA retrieval). Frozen sections are not shown to be easier; HPA provides ICC-IF images in MCF-7 and RT-4 cells, while kidney sections using biotin-based detection need an endogenous-biotin check (HPA: subcellular ICC-IF; caption: SABC; standard IHC practice).
HPA tissue IHC evidence for ABCC5
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low consistency between antibody staining and RNA expression data. Pending retesting.). 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 |
|---|---|---|---|---|
| Appendix | Lymphoid tissue | Medium | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Medium | Protein (IHC) | HPA → |
| Kidney | Cells in tubules | Medium | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Medium | Protein (IHC) | HPA → |
| Placenta | Trophoblastic cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ABCC5 IHC Tips
Troubleshooting ABCC5 chromogenic IHC in paraffin sections requires attention to retrieval, membrane localisation and the limits of the available staining evidence.
What retrieval should I try first when ABCC5 staining is weak?
Start with heat mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A03040-1). The selected tissue image used that retrieval before incubation with 2 μg/ml primary antibody overnight at 4°C, so keep those conditions together during the first comparison (datasheet A03040-1). If staining remains weak, compare a shorter and longer heating exposure on matched sections while keeping the antibody concentration and detection steps fixed (standard IHC practice). Score membrane associated staining separately from diffuse signal, because ABCC5 has 13 transmembrane segments and documented plasma membrane localisation (UniProt O15440 topology; HPA subcellular).
Could fixation explain inconsistent ABCC5 staining between paraffin blocks?
Target specific ABCC5 sensitivity to fixation is unknown: the selected paraffin tissue caption does not state a fixative (datasheet A03040-1). Record each block’s fixative and fixation duration, then compare sections with the same EDTA pH 8.0 retrieval and 2 μg/ml primary antibody conditions (datasheet A03040-1; standard IHC practice). Uneven preservation, section detachment or tissue morphology can help identify processing differences before assigning a staining difference to ABCC5 abundance (standard IHC practice). Use a matched control section in every run, and avoid inferring fixation tolerance from tissue staining patterns or predicted protein features (standard IHC practice).
How should I evaluate cytoplasmic staining when ABCC5 is a membrane transporter?
Examine whether chromogen outlines a cell surface or junction, and record diffuse cytoplasmic staining as a separate pattern (HPA subcellular; standard IHC practice). UniProt places ABCC5 at basolateral and apical membranes, endosome membranes, cytoplasmic granules and the Golgi apparatus lumen; it describes apical localisation in brain capillary endothelial cells (UniProt O15440 subcellular). HPA reports mainly plasma membrane and cell junction localisation, with additional nucleoplasmic signal, while its tissue IHC profile describes cytoplasmic expression (HPA subcellular; HPA tissue IHC). Because HPA rates its tissue staining reliability uncertain, require reproducible compartment patterns and appropriate controls before interpreting cytoplasmic DAB as ABCC5 (HPA tissue IHC; standard IHC practice).
Could isoforms or epitope accessibility change the IHC pattern?
ABCC5 has 5 annotated isoforms, but the supplied antibody caption does not identify its epitope or establish which isoforms it detects (UniProt O15440 isoforms; datasheet A03040-1). Its 13 transmembrane segments and 8 annotated glycosylation sites make epitope position relevant when assessing access after paraffin processing and retrieval (UniProt O15440 topology and glycosylation; standard IHC practice). Request the immunogen or mapped epitope before attributing different staining patterns to isoform expression (standard IHC practice). Meanwhile, compare serial sections under the documented EDTA pH 8.0 retrieval conditions and describe the observed compartment without assigning it to a particular isoform (datasheet A03040-1; standard IHC practice).
How can IF help assess an uncertain ABCC5 IHC pattern?
Use IF as a separate localisation check and multiplex ABCC5 with a marker of the expected cell type, such as a marker identifying kidney tubular cells when examining kidney (HPA tissue IHC; standard IF practice). Choose a far red fluorophore where tissue autofluorescence obscures shorter wavelengths, and inspect single stain controls before interpreting overlap (standard IF practice). Match permeabilisation to the antibody’s mapped epitope: access to a cytoplasmic epitope may require it, whereas a surface accessible epitope may be assessed without it (standard IF practice). HPA lists IF images in MCF-7 and RT-4 and reports plasma membrane and junction signal; those observations do not establish this catalog antibody’s IF performance (HPA subcellular).
How do I separate ABCC5 staining from detection background?
The selected paraffin tissue image used 10% goat serum blocking, a biotinylated secondary antibody, streptavidin biotin detection and DAB (datasheet A03040-1). Run a no primary control through the same detection steps to reveal background from the secondary, endogenous biotin or endogenous peroxidase (standard IHC practice). Include a peroxidase block before DAB development and assess whether residual colour follows tissue structures or section edges rather than plausible ABCC5 compartments (standard IHC practice; UniProt O15440 subcellular). If background persists, optimise blocking and washing while retaining EDTA pH 8.0 retrieval as the documented starting condition (datasheet A03040-1; standard IHC practice).
What should I score when comparing ABCC5 across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring; keep membrane associated and diffuse cytoplasmic DAB measurements separate (HPA subcellular; HPA tissue IHC; standard IHC practice). An H-score combines intensity with the percentage of positive cells, while percentage positive alone may be easier to reproduce when intensity calibration is uncertain (standard IHC practice). Normalise each score to the number of evaluable cells in the chosen compartment, or report positive cell density per mm² of viable tissue when cell counting is impractical (standard IHC practice). Apply identical thresholds, retrieval and detection settings across sections, and report HPA’s uncertain tissue IHC reliability alongside the results (datasheet A03040-1; HPA tissue IHC).
When is an ABCC5 positive IHC result credible?
Look for reproducible staining in a plausible cell population and compartment, supported by a clean no primary control (UniProt O15440 subcellular; HPA subcellular; standard IHC practice). Kidney tubular cells and placental trophoblastic cells have reported medium tissue IHC staining, while adipocytes have a reported not detected result; these are comparison points, not definitive controls (HPA tissue IHC). Treat strong colour confined to torn edges, necrotic areas or sites of endogenous enzyme activity as a potential artefact (standard IHC practice). Interpret isolated nuclear or diffuse cytoplasmic signal cautiously: HPA reports additional nucleoplasmic and cytoplasmic patterns, but rates tissue IHC reliability uncertain because staining and RNA expression show low consistency (HPA subcellular; HPA tissue IHC).
Boster reagents
Best ABCC5 / ATP-binding cassette sub-family C member 5 IHC Antibodies
The catalog lists one human-reactive anti-ABCC5 antibody with real IHC data from paraffin-embedded human gastric and mammary cancer sections (catalog: A03040-1 IHC captions).
Real IHC data
Anti-MRP5/ABCC5 Antibody
Cat # A03040-1
A03040-1 is listed for human IHC, flow cytometry and ELISA (catalog: A03040-1 applications and reactivity). Its IHC captions show staining in paraffin-embedded human gastric and mammary cancer sections (catalog: A03040-1 IHC captions).
Which to pick: Choose A03040-1 for human paraffin-section IHC; it is a rabbit polyclonal antibody with an IHC application listing and matching tissue images (catalog: A03040-1 applications, reactivity, clonality and IHC captions). The captions report paraffin embedding but do not report the fixative (catalog: A03040-1 IHC captions). No listed SKU has IF/ICC validation or cross-species reactivity (catalog: A03040-1 applications and reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O15440 (MRP5_HUMAN, ATP-binding cassette sub-family C member 5).
- Human Protein Atlas. ABCC5 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. ABCC5 subcellular location (ICC-IF): Mainly localized to the plasma membrane and cell junctions. In addition localized to the nucleoplasm..
- Human Protein Atlas. ABCC5 antibody validation summary (2 antibodies).
- Loss of miR-516a-3p mediates upregulation of ABCC5 in prostate cancer and drives its progression. OncoTargets and therapy 2018 — PMC6039062.
- Identification and Validation of Three-Gene Signature in Lung Squamous Cell Carcinoma by Integrated Transcriptome and Methylation Analysis. Journal of oncology 2022 — PMC9525794.
- Three novel ABCC5 splice variants in human retina and their role as regulators of ABCC5 gene expression. BMC molecular biology 2007 — PMC1890297.
- ABCC5 supports osteoclast formation and promotes breast cancer metastasis to bone. Breast cancer research : BCR 2012 — PMC4053136.
- PubMed PMID:9827529 — UniProt-cited evidence.
- PubMed PMID:10438534 — UniProt-cited evidence.
- PubMed PMID:10721709 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ABCC5 (ATP-binding cassette sub-family C member 5). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A03040-1). Validated IHC antibody: A03040-1 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Lymphoid tissue); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human gastric cancer tissue; fixative not specified (datasheet A03040-1); 2. Fixation: Image fixative and duration unreported (datasheet A03040-1); 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 retrieval: EDTA pH 8.0 (datasheet A03040-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03040-1); 8. Primary antibody: Rabbit anti-ABCC5, 2-5μg/ml (datasheet A03040-1); 9. Primary incubation: Overnight at 4 °C (datasheet A03040-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific). Selected-image fixative and duration unreported (datasheet A03040-1); verify before use.; Caveat: Staining and RNA levels show low consistency (HPA tissue IHC); Isoform / epitope: 5 isoforms; extracellular versus cytoplasmic epitope matters (UniProt); Regulation: Low tissue specificity (HPA tissue IHC). Evidence: UniProt O15440, HPA tissue IHC · Uncertain, HPA ICC-IF · Mainly localized to the plasma membrane and cell junctions. In…, Datasheet A03040-1, PMC4053136. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ABCC5/immunohistochemistry-ihc-guide.