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ABCC3 / ATP-binding cassette sub-family C member 3 · IHC design guide
Design Immunohistochemistry for ABCC3
Plan ABCC3 chromogenic IHC on paraffin sections using glandular cells in appendix or colon as high-staining references (HPA tissue IHC). This guide covers fixation, interpretation of membranous and cytoplasmic staining, and the catalog antibody’s 2–5 μg/ml IHC range (datasheet A02429-1).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ABCC3 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 | Membranous and cytoplasmic (HPA tissue IHC); basolateral membrane (UniProt) | |
| Staining pattern | Glandular cells show membranous and cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A02429-1) | |
| 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 paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Cytoplasmic signal may complicate basolateral scoring (HPA tissue IHC) | |
| Regulation | No target-specific induction is annotated (UniProt) | |
| Isoform / epitope | 5 isoforms; map the epitope to each variant and membrane face (UniProt) |
Section 1
Recommended ABCC3 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet A02429-1) with a published tumor tissue IHC protocol (PMC11450052: methods).
| Sample | Paraffin-embedded human appendiceal adenocarcinoma tissue; fixative not specified (datasheet A02429-1) |
| Fixation | Image fixative and duration unreported (datasheet A02429-1); 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 A02429-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02429-1) |
| Primary antibody | Rabbit anti-ABCC3, 2-5 μg/ml (datasheet A02429-1) |
| Primary incubation | Overnight at 4 °C (datasheet A02429-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A02429-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ABCC3-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Membranous and cytoplasmic expression in several tissues including adrenal gland, gastrointestinal tract and gallbladder. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet A02429-1); the published tumor protocol used citrate pH 6.0 (PMC11450052: methods).
Section 2
What Is the Expected ABCC3 Staining Pattern?
ABCC3 is a 17-pass membrane transporter expected at basal or basolateral cell membranes (UniProt O15438 topology and subcellular location). In paraffin-section IHC, assess glandular cells in adrenal gland, colon, appendix, gallbladder and rectum, where HPA reports high staining (HPA tissue IHC). HPA also describes membranous and cytoplasmic staining across several tissues; its tissue IHC reliability is Enhanced, with medium consistency against RNA data (HPA tissue IHC).
| Distinct membrane-edge staining in glandular cells, especially along a basal or basolateral border. | This fits ABCC3's annotated membrane location and polarized-cell role (UniProt O15438 subcellular location and function). High glandular-cell staining in adrenal gland, appendix, colon, gallbladder or rectum provides a tissue context for the result (HPA tissue IHC). Interpret the cell border together with tissue structure; a dark edge alone does not establish antibody specificity (general IHC practice). |
| Cytoplasmic staining accompanies a discernible membrane pattern in the expected cells. | Cytoplasmic signal is included in HPA's tissue IHC profile, so it does not by itself invalidate a section (HPA tissue IHC). A predominantly nuclear pattern, or staining confined to a compartment unrelated to the reported membrane and cytoplasmic pattern, is discordant and warrants review (HPA tissue IHC; UniProt O15438 subcellular location). Check morphology and controls before calling it an artefact (general IHC practice). |
| Strong staining appears in cells outside the expected glandular-cell pattern, including a negative-control cell population. | For example, HPA reports no detection in adipocytes of adipose tissue or hematopoietic cells of bone marrow (HPA tissue IHC). Signal there raises concern for cross-reactivity or endogenous detection activity; its appearance cannot identify the cause on its own (general IHC practice). Compare a no-primary control, cell morphology and staining in a known-positive tissue (general IHC practice). |
| Color is spread broadly across the section, including tissue spaces or many unrelated cell types. | A diffuse wash of chromogen is less persuasive than a cell-associated membrane pattern (UniProt O15438 subcellular location; general IHC practice). It may reflect nonspecific staining or detection background, but appearance alone cannot assign a single cause (general IHC practice). Inspect the no-primary control and whether the signal tracks tissue structure before scoring ABCC3 (general IHC practice). |
| No signal is visible in glandular cells of a tissue HPA reports as highly stained. | An absent result in adrenal gland, colon, appendix, gallbladder or rectum conflicts with the reported high glandular-cell staining (HPA tissue IHC). First check that the expected cells are present and that a positive control stained in the same run (general IHC practice). The result may reflect a run-level failure or specimen variability; it does not alone establish that ABCC3 is absent (general IHC practice). |
💡Expected ABCC3 appearanceCall a positive result when glandular cells show convincing membrane-associated staining, preferably at a basal or basolateral edge, with cytoplasmic staining allowable (UniProt O15438 subcellular location; HPA tissue IHC); high staining is reported in selected glandular tissues (HPA tissue IHC), whereas isolated nuclear or widespread cell-independent color is suspect (general IHC practice).
| Membrane topology and polarity (UniProt O15438 topology and subcellular location) | ABCC3 has 17 transmembrane segments and is annotated at basal or basolateral membranes (UniProt O15438). Score membrane position in intact cells when tissue orientation permits; do not require every cut cell to show a complete outline (general IHC practice). |
| Tissue and cell-type context (HPA tissue IHC) | HPA reports high staining in glandular cells of several tissues but no detection in liver cholangiocytes (HPA tissue IHC). The cholangiocyte entry does not describe hepatocytes; UniProt separately reports predominant liver expression (UniProt O15438 tissue specificity). Keep those cell populations distinct when choosing or interpreting controls. |
| Antibody evidence (HPA antibody validation; HPA tissue IHC) | HPA048483 is IHC Enhanced and CAB037136 is IHC Approved (HPA antibody validation). The overall tissue IHC profile is Enhanced but has medium consistency with RNA data (HPA tissue IHC). Treat a matching pattern as supported evidence, while retaining tissue and run controls for any particular slide (general IHC practice). |
| Isoforms and epitope scope (UniProt O15438 isoforms and topology) | UniProt lists five ABCC3 isoforms and extracellular and cytoplasmic regions (UniProt O15438). These facts alone do not show which isoforms an antibody detects or how a paraffin-section epitope behaves. Check the selected antibody's documented epitope and IHC validation before interpreting an unexpected compartment (general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected glandular cells are negative in an HPA high-staining tissue (HPA tissue IHC). | The expected cells may be absent from the section, or the IHC run may have failed (general IHC practice). | Confirm cell identity and section quality; review a positive control processed in the same run and the antibody's IHC instructions (general IHC practice). Do not infer target-specific fixation sensitivity from this result. |
| The section shows widespread color with weak cell boundaries. | Nonspecific staining or detection background can obscure a membrane pattern (general IHC practice). | Compare the no-primary control, blocking and detection controls; adjust reagent conditions according to the validated IHC method (general IHC practice). Reassess whether signal follows glandular-cell membranes (UniProt O15438; HPA tissue IHC). |
| Nuclei stain more strongly than cell borders. | Nuclear-dominant staining is discordant with reported membrane and cytoplasmic localization (UniProt O15438 subcellular location; HPA tissue IHC). | Check a no-primary control and a positive tissue in the same run, then review the antibody's IHC validation before assigning ABCC3 positivity (general IHC practice). |
| Unexpected cells stain strongly while the expected glandular cells do not. | Cross-reactivity or endogenous detection activity is possible; the slide alone cannot distinguish them (general IHC practice). | Compare matched controls and cell morphology, especially in an HPA-reported high-staining tissue and a reported undetected population (HPA tissue IHC; general IHC practice). |
| A liver section appears negative in cholangiocytes. | HPA reports ABCC3 as not detected in liver cholangiocytes, while UniProt describes predominant liver expression without making that a cholangiocyte claim (HPA tissue IHC; UniProt O15438 tissue specificity). | Identify hepatocytes and cholangiocytes separately before judging the liver result; use a documented glandular-cell positive tissue to assess the run (HPA tissue IHC; general IHC practice). |
| IF/ICC shows plasma-membrane signal plus a basal-body focus: is that expected? | HPA supports plasma-membrane localization in ICC-IF but marks the additional basal-body location uncertain (HPA subcellular ICC-IF). | Use the plasma-membrane pattern as the stronger localization reference and treat a basal-body focus as provisional (HPA subcellular ICC-IF). Evaluate IF/ICC on its dedicated guide page; it does not provide an IHC protocol option here. |
Experiment evidence
IHC & IF images showing ABCC3 staining
A02429-1 · IHCIHC analysis of MRP3/ABCC3 using anti-MRP3/ABCC3 antibody (A02429-1). MRP3/ABCC3 was detected in a paraffin-embedded section of human append
A02429-1 · IHCIHC analysis of MRP3/ABCC3 using anti-MRP3/ABCC3 antibody (A02429-1). MRP3/ABCC3 was detected in a paraffin-embedded section of human coloni
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000108846-ABCC3/tissue/cerebral+cortex#imid_16738943
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000108846-ABCC3/tissue/colon#imid_16744089
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000108846-ABCC3/tissue/liver#imid_16744082
HPA · GallbladderHuman Protein Atlas IHC, Gallbladder. Source: https://www.proteinatlas.org/ENSG00000108846-ABCC3/tissue/gallbladder#imid_16744074
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA048483; green = ABCC3, blue = nucleus, red = microtubules.
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA048483; green = ABCC3, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ABCC3 IHC & IF
🧪Run appendix first: its glandular cells should stain (HPA: High in appendix glandular cells). Run adipose tissue as the negative, focusing on adipocytes (HPA: Not detected in adipocytes); on the appendix slide, assess adjacent nonglandular cells for background without assuming they are ABCC3-negative (HPA: appendix annotation specifies glandular cells).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ABCC3 in A-549, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Plasma membrane (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and concentration-matched rabbit IgG isotype controls, plus ABCC3-knockout tissue or a validated peptide-block control (datasheet: rabbit primary antibody; standard IHC practice). For the appendix slide, check endogenous peroxidase and biotin background with the chromogenic detection system used in the caption; for IF, check tissue autofluorescence (caption: biotinylated secondary, streptavidin–biotin complex and DAB; standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A02429-1 paraffin-section caption does not state the fixative (selected-SKU caption). The demonstrated IHC procedure uses heat retrieval in EDTA pH 8.0, so assess staining with that retrieval condition; the caption does not establish that retrieval is required under every processing condition (selected-SKU caption). Paraffin IHC has a supplied example, while the evidence does not establish that frozen sections or IF are easier; in appendix, evaluate DAB signal against endogenous peroxidase and biotin background (selected-SKU caption; standard IHC practice).
HPA tissue IHC evidence for ABCC3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | 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 → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ABCC3 IHC Tips
Use the catalog antibody’s paraffin-section protocol as the starting point, then judge ABCC3 staining by cell type, membrane position, and controls.
What retrieval should I try when ABCC3 staining is weak in paraffin sections?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 (datasheet A02429-1). The catalog image used a paraffin-embedded human appendiceal adenocarcinoma section, followed by 10% goat serum blocking and 2 μg/ml primary antibody overnight at 4°C (datasheet A02429-1). If staining remains weak, compare a second retrieval condition on adjacent sections while keeping antibody concentration, detection, and exposure to chromogen constant (standard IHC practice). Judge the result by preserved morphology and convincing basolateral or basal membrane staining, rather than total brown signal alone (UniProt O15438 subcellular location; standard IHC practice).
How should I troubleshoot variable ABCC3 staining across differently fixed sections?
The selected paraffin-section caption does not state its fixative, so ABCC3-specific fixation sensitivity is unknown for this antibody (datasheet A02429-1). Record each specimen’s fixative and fixation duration, and compare sections processed with matched retrieval and detection conditions before changing antibody concentration (standard IHC practice). Use the stated EDTA pH 8.0 heat retrieval as the initial condition for this catalog antibody (datasheet A02429-1). If morphology or staining varies, assess matched positive and negative tissue controls alongside the samples; neither tissue-expression patterns nor protein topology establish a fixation effect (HPA tissue IHC; UniProt O15438 topology; standard IHC practice).
Where should convincing ABCC3 staining appear in polarized tissue?
Prioritize basolateral or basal membrane staining in appropriately polarized cells: ABCC3 is annotated at these membrane domains, including the basolateral membrane of enterocytes (UniProt O15438 subcellular location; PubMed:28408210). HPA also reports membranous and cytoplasmic staining across several tissues, so document cytoplasmic signal separately rather than treating it as equivalent to membrane localization (HPA tissue IHC). Compare the membrane facing the underlying tissue with the luminal border in intact glands, using morphology and a counterstain to orient each cell (UniProt O15438 subcellular location; standard IHC practice). Diffuse staining without a discernible cellular pattern warrants review of retrieval, background, and section integrity (standard IHC practice).
Could epitope placement or alternative splicing explain inconsistent ABCC3 staining?
ABCC3 has 5 annotated isoforms and 17 transmembrane segments, so epitope location matters when interpreting an antibody’s staining pattern (UniProt O15438 isoforms and topology). The supplied catalog caption does not identify the antibody epitope; do not assign isoform coverage or an extracellular versus cytoplasmic binding site from that image (datasheet A02429-1). Consult the antibody’s mapped immunogen or epitope information before comparing it with isoform sequences or choosing permeabilization conditions (standard IHC practice). For discordant sections, compare the same cell compartment across specimens and report the antibody identity and retrieval condition with the result (UniProt O15438 subcellular location; standard IHC practice).
How can I use IF to investigate an ambiguous chromogenic ABCC3 pattern?
Use IF as a complementary localization check for an ambiguous IHC pattern, pairing ABCC3 with a marker that identifies the expected cell population in the specimen (standard IF practice). Choose fluorophores and filter sets after checking tissue autofluorescence and include single-label controls for multiplex imaging (standard IF practice). ABCC3 spans the membrane 17 times and has extracellular and cytoplasmic regions; choose permeabilization only after establishing which side contains the antibody epitope (UniProt O15438 topology; standard IF practice). Compare any IF membrane pattern with the IHC section’s basolateral or basal staining, without assuming that the paraffin-section antibody conditions validate IF (UniProt O15438 subcellular location; datasheet A02429-1).
What should I adjust when DAB obscures ABCC3 membrane staining?
Check a no-primary control to identify signal from the secondary reagent, detection system, or endogenous tissue activity (standard IHC practice). The catalog image used a biotinylated secondary antibody, streptavidin–biotin detection, and DAB, making detection-system background relevant to this workflow (datasheet A02429-1). Apply an appropriate endogenous peroxidase block, assess background from the biotin-based system, and shorten DAB development if broad brown deposits mask cell borders (standard IHC practice). Compare the adjusted section with a matched positive control and favor a resolved basolateral membrane pattern over an increase in overall staining intensity (UniProt O15438 subcellular location; standard IHC practice).
How should I score ABCC3 when membrane and cytoplasmic staining differ? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored compartment before analysis, separating basolateral or basal membrane staining from cytoplasmic staining (UniProt O15438 subcellular location; HPA tissue IHC). Within the same annotated cell population, report the percentage of positive cells and a membrane-specific H-score; record cytoplasmic staining separately if it is biologically relevant to the question (standard IHC practice). Normalize cell-based scores to the number of evaluable target cells, or normalize stained area or cell counts to evaluable tissue area in mm² (standard IHC practice). Keep retrieval, chromogen development, image acquisition, thresholds, and exclusion rules consistent across specimens (standard IHC practice).
How can I distinguish true ABCC3 staining from tissue artefact?
A credible ABCC3 pattern follows the expected cell population and favors basolateral or basal membrane localization over indiscriminate luminal, nuclear, or extracellular staining (UniProt O15438 subcellular location; standard IHC practice). HPA reports high staining in appendix and colon glandular cells, while its tissue profile also includes cytoplasmic signal; use these observations as context rather than proof for a new specimen (HPA tissue IHC). Review section edges, folds, and necrotic regions for localized staining artefacts, and inspect a no-primary control for endogenous enzyme or detection-system signal (standard IHC practice). Require agreement between morphology, compartment, and controls before assigning a positive score (standard IHC practice).
Boster reagents
Best ABCC3 / ATP-binding cassette sub-family C member 3 IHC Antibodies
A02429-1 has IHC images from human paraffin sections (catalog IHC captions); the catalog supplies no IF/ICC validation or images (catalog applications and IF image alts).
Real IHC data
Anti-MRP3/ABCC3 Antibody ®
Cat # A02429-1
A02429-1 is listed for human IHC (catalog applications and reactivity). Its IHC captions show paraffin sections of human appendiceal adenocarcinoma, colonic adenocarcinoma, placenta and stomach cancer (catalog IHC captions).
Which to pick: For tissue IHC, choose A02429-1, a human-reactive rabbit polyclonal antibody with a listed IHC dilution of 2–5 μg/ml and paraffin-section images; the fixative is unreported (catalog reactivity, dilution and IHC captions). For IF/ICC, no SKU has listed validation or IF images (catalog applications and IF image alts). For other species, no listed SKU has documented reactivity beyond human (catalog reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O15438 (MRP3_HUMAN, ATP-binding cassette sub-family C member 3).
- Human Protein Atlas. ABCC3 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. ABCC3 subcellular location (ICC-IF): Mainly localized to the plasma membrane. In addition localized to the basal body..
- Human Protein Atlas. ABCC3 antibody validation summary (2 antibodies).
- Quercetin Regulates Key Components of the Cellular Microenvironment during Early Hepatocarcinogenesis. Antioxidants (Basel, Switzerland) 2022 — PMC8868318.
- Detection of ABCC1 expression in classical Hodgkin lymphoma is associated with increased risk of treatment failure using standard chemotherapy protocols. Journal of hematology & oncology 2012 — PMC3470996.
- Downregulation of ABCC3 activates MAPK signaling through accumulation of deoxycholic acid in colorectal cancer cells. Cancer science 2024 — PMC11145118.
- Molecular mechanisms and diagnostic model of glioma-related epilepsy. NPJ precision oncology 2024 — PMC11450052.
- PubMed PMID:9813153 — UniProt-cited evidence.
- PubMed PMID:9738950 — UniProt-cited evidence.
- PubMed PMID:9827529 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ABCC3 (ATP-binding cassette sub-family C member 3). Expected localisation Membranous and cytoplasmic (HPA tissue IHC); basolateral membrane (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A02429-1). Validated IHC antibody: A02429-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human appendiceal adenocarcinoma tissue; fixative not specified (datasheet A02429-1); 2. Fixation: Image fixative and duration unreported (datasheet A02429-1); 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 A02429-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02429-1); 8. Primary antibody: Rabbit anti-ABCC3, 2-5 μg/ml (datasheet A02429-1); 9. Primary incubation: Overnight at 4 °C (datasheet A02429-1). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Cytoplasmic signal may complicate basolateral scoring (HPA tissue IHC); Isoform / epitope: 5 isoforms; map the epitope to each variant and membrane face (UniProt); Regulation: No target-specific induction is annotated (UniProt). Evidence: UniProt O15438, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the plasma membrane. In addition localized to…, Datasheet A02429-1, PMC11450052. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ABCC3/immunohistochemistry-ihc-guide.