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MFSD2A / Sodium-dependent lysophosphatidylcholine symporter 1 · IHC design guide
Design Immunohistochemistry for MFSD2A
Plan MFSD2A chromogenic IHC on paraffin sections using the observed cytoplasmic tissue profile (HPA tissue IHC) and annotated membrane and ER localisation (UniProt). Use the reported high staining in appendix, breast, or salivary gland glandular cells as a reference, while accounting for the uncertain reliability of the tissue staining (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MFSD2A 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); membrane and ER localisation (UniProt) | |
| Staining pattern | Cytoplasmic staining in most tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A01967-2) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Heart muscle+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 has low consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Tissue-enhanced RNA expression (HPA tissue RNA) | |
| Isoform / epitope | 3 isoforms; check extracellular versus cytoplasmic epitope (UniProt) |
Section 1
Recommended MFSD2A IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: A01967-2) is accompanied by published paraffin-section workflows for MFSD2A (PMC2846890; PMC6814593).
| Sample | Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A01967-2) |
| Fixation | Image fixative and duration unreported (datasheet A01967-2); 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 A01967-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01967-2) |
| Primary antibody | Rabbit anti-MFSD2A, 2-5 μg/ml (datasheet A01967-2) |
| Primary incubation | Overnight at 4 °C (datasheet A01967-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A01967-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | MFSD2A-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet: A01967-2); citrate pH 6 is a published alternative (PMC2846890).
Section 2
What Is the Expected MFSD2A Staining Pattern?
MFSD2A is a 12-pass membrane protein assigned to the cell and endoplasmic reticulum membranes, with possible cytoplasmic puncta (UniProt Q8NA29 topology and subcellular location). In tissue IHC, HPA reports mainly cytoplasmic staining, including high staining in selected glandular cells and cerebellar Purkinje cells (HPA: tissue IHC). Treat this as a provisional pattern: HPA rates its tissue staining Uncertain because antibody staining and RNA expression have low consistency (HPA: reliability).
| Strong cytoplasmic staining in appendix, breast, or salivary gland glandular cells, or in cerebellar Purkinje cells (HPA: High in these cells). | This matches reported tissue staining. A membrane-associated component is biologically plausible for a transmembrane transporter, but the tissue IHC profile is described as cytoplasmic; do not require a crisp membrane outline to call the pattern concordant (UniProt Q8NA29 topology; HPA: tissue IHC). |
| Predominantly nuclear staining, with little cytoplasmic or membrane-associated signal (HPA: cytoplasmic tissue profile; UniProt Q8NA29 subcellular location). | Nuclear dominance conflicts with the supplied localisation evidence. Consider nonspecific staining or a detection artefact, and compare with a no-primary control before assigning the signal to MFSD2A (HPA: tissue IHC; UniProt Q8NA29 subcellular location; general IHC practice). |
| Strong staining in cardiomyocytes, skeletal myocytes, or prostate glandular cells (HPA: Not detected in these cells). | This disagrees with HPA's reported negatives and raises cross-reactivity or endogenous detection activity as possibilities. HPA's overall IHC reliability is Uncertain, so investigate the discrepancy rather than treating those cells as a definitive biological negative (HPA: tissue IHC and reliability; general IHC practice). |
| Diffuse chromogen across many cell types and tissue spaces, without clear cellular boundaries (general IHC practice). | A widespread haze cannot establish MFSD2A localisation. Check the no-primary control and detection background, then reassess whether any cell-associated staining remains; HPA's broad cytoplasmic profile alone cannot validate diffuse signal (general IHC practice; HPA: tissue IHC and reliability). |
| No staining in an appendix, breast, or salivary gland section where the sampled glandular cells are present (HPA: High in glandular cells). | The result differs from HPA's reported pattern, but those tissues are provisional comparison material because HPA rates IHC reliability Uncertain. Check tissue preservation, retrieval, antibody and detection performance, and a concurrent positive control before interpreting absence as biological (HPA: tissue IHC and reliability; general IHC practice). |
💡Expected MFSD2A appearanceA plausible positive is cell-associated cytoplasmic staining, potentially with membrane-associated signal, in HPA-listed glandular cells or Purkinje cells at the reported High level; dominant nuclear staining or uniform signal in HPA-listed negative cells warrants investigation (HPA: tissue IHC and reliability; UniProt Q8NA29 subcellular location).
| Membrane topology (UniProt Q8NA29 topology) | MFSD2A has 12 transmembrane segments and both cytoplasmic and extracellular regions. Epitope position could affect antibody access, but the supplied record does not locate the IHC antibody's epitope or establish a retrieval requirement (UniProt Q8NA29 topology; general IHC practice). |
| Subcellular distribution (UniProt Q8NA29 subcellular location; HPA: subcellular ICC-IF) | UniProt places MFSD2A at cell and endoplasmic reticulum membranes and notes possible cytoplasmic puncta. HPA ICC-IF supports plasma-membrane localisation, calls cytosol uncertain, and lists cytoplasmic bodies as an additional approved location; these observations do not set an IHC scoring threshold (UniProt Q8NA29 subcellular location; HPA: subcellular ICC-IF). |
| Tissue evidence and antibody validation (HPA: tissue IHC and antibodies) | HPA reports high staining in selected cells but rates the overall tissue profile Uncertain; antibody HPA043107 is also marked IHC Uncertain. Use concordant cell and compartment patterns as supporting evidence, not proof of target specificity (HPA: tissue IHC, reliability, and antibodies). |
| Isoforms and glycosylation (UniProt Q8NA29 isoforms and glycosylation) | UniProt lists 3 isoforms and glycosylation sites at residues 230 and 240. The payload does not show which isoforms the IHC antibody recognises or whether these sites alter staining, so neither feature predicts a specific positive or negative section (UniProt Q8NA29 isoforms and glycosylation). |
| Situation | Likely cause | Next action |
|---|---|---|
| The expected glandular cells or Purkinje cells have no visible signal (HPA: High in those cells). | The assay may have failed, the sampled cells may be absent, or this section may differ from HPA's uncertain profile (HPA: tissue IHC and reliability; general IHC practice). | Confirm the relevant cells are present; review retrieval, antibody dilution and chromogenic detection against the catalog IHC-P instructions; include a concurrent positive control. Do not infer a target-specific fixation effect from this result alone (general IHC practice; HPA: reliability). |
| The entire section shows diffuse brown background (general IHC practice). | Nonspecific antibody binding or endogenous detection activity can obscure cell-associated staining (general IHC practice). | Inspect no-primary and detection controls, then review blocking, washes and antibody dilution. Score MFSD2A only after distinct cellular staining can be separated from background (general IHC practice). |
| Nuclei dominate the stain while cytoplasm is faint (HPA: cytoplasmic tissue profile). | The pattern conflicts with UniProt membrane localisation and HPA tissue staining; its cause cannot be assigned from morphology alone (UniProt Q8NA29 subcellular location; HPA: tissue IHC). | Compare with a no-primary control and an HPA-listed high-staining cell population on the same or a parallel section. Record persistent nuclear signal as discordant rather than counting it as MFSD2A-positive (general IHC practice; HPA: tissue IHC). |
| HPA-listed negative cells stain as strongly as the intended comparison cells (HPA: tissue IHC). | Cross-reactivity or endogenous detection activity is possible; HPA's uncertain reliability also limits how decisively its negative calls can be used (HPA: tissue IHC and reliability; general IHC practice). | Check no-primary and detection controls, compare staining by cell type and compartment, and seek independent target validation before interpreting the unexpected cells as expressing MFSD2A (general IHC practice; HPA: reliability). |
| Membrane edges and cytoplasmic puncta appear together (UniProt Q8NA29 subcellular location). | Both can fit the reported cell-membrane and possible vesicular distribution, although morphology alone cannot identify the puncta (UniProt Q8NA29 subcellular location). | Score each compartment separately and compare the stained cell types with HPA tissue IHC. Avoid calling every punctum specific because the HPA tissue profile remains Uncertain (HPA: tissue IHC and reliability; general IHC practice). |
| Can ICC-IF localisation settle an ambiguous chromogenic IHC result? (HPA: subcellular ICC-IF and tissue IHC) | HPA supports plasma-membrane localisation by ICC-IF, while its tissue IHC profile and the listed antibody's IHC validation are Uncertain; the assays provide different evidence (HPA: subcellular ICC-IF, tissue IHC, and antibodies). | Use the separate IF/ICC guide for that application. Here, compare IHC cell types, compartments and controls, and report any disagreement with the ICC-IF localisation explicitly (HPA: subcellular ICC-IF and tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing MFSD2A staining
A01967-2 · IHCIHC analysis of MFSD2A using anti-MFSD2A antibody (A01967-2). MFSD2A was detected in a paraffin-embedded section of human liver cancer tissu
A01967-2 · IHCIHC analysis of MFSD2A using anti-MFSD2A antibody (A01967-2). MFSD2A was detected in a paraffin-embedded section of human liver tissue. Heat
A01967-2 · IFIF analysis of MFSD2A using anti-MFSD2A antibody (A01967-2). MFSD2A was detected in an immunocytochemical section of SIHA cells. Enzyme anti
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000168389-MFSD2A/tissue/cerebral+cortex#imid_11323895
HPA · CerebellumHuman Protein Atlas IHC, Cerebellum. Source: https://www.proteinatlas.org/ENSG00000168389-MFSD2A/tissue/cerebellum#imid_11323882
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000168389-MFSD2A/tissue/colon#imid_11323821
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000168389-MFSD2A/tissue/liver#imid_11323806
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA043107; green = MFSD2A, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA043107; green = MFSD2A, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MFSD2A IHC & IF
🧪Run appendix first and look for staining in glandular cells (HPA: High in appendix glandular cells). Use skeletal muscle myocytes as the negative tissue (HPA: Not detected); on the appendix slide, assess non-glandular cells for background staining, recognizing that the HPA row establishes a level only for glandular cells.
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: Heart muscle (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MFSD2A in A-431, U-251MG, U2OS, with annotated localisation: Plasma membrane (supported), Cytosol (uncertain) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a matched nonimmune rabbit IgG control, using the appropriate monoclonal isotype or polyclonal IgG for the primary antibody (A01967-2 caption: rabbit primary). Use an MFSD2A knockout specimen as a biological negative; for chromogenic detection, quench endogenous peroxidase and check whether inflammatory cells in appendix contribute background (A01967-2 caption: HRP/DAB detection).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A01967-2 paraffin-section caption does not report a fixative. The caption uses heat retrieval in EDTA at pH 8.0, but whether retrieval is required is unreported (A01967-2 caption). The supplied evidence does not establish whether frozen sections or IF are easier; appendix inflammatory cells may complicate interpretation of chromogenic background (A01967-2 caption: HRP/DAB detection).
HPA tissue IHC evidence for MFSD2A
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low consistency between antibody staining and RNA expression data. Pending external verification.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Salivary gland | Glandular cells | High | Protein (IHC) | HPA → |
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Prostate | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MFSD2A IHC Tips
Troubleshoot MFSD2A chromogenic IHC in paraffin sections using documented retrieval conditions, compartment patterns, cellular context, and staining controls.
What retrieval conditions should I start with for weak MFSD2A staining in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A01967-2). The selected tissue-IHC image used this condition, followed by 10% goat-serum blocking and 2 μg/ml primary antibody overnight at 4°C (datasheet A01967-2). If staining remains weak, vary heating time in small increments while holding section thickness, antibody concentration, and detection constant (standard IHC practice). Inspect morphology and background alongside signal, because excessive heating can damage tissue and increase nonspecific staining (standard IHC practice). Compare each trial with a no-primary control and a consistently processed reference section before changing the retrieval buffer (standard IHC practice).
Could fixation explain inconsistent MFSD2A staining between paraffin blocks?
MFSD2A-specific fixation sensitivity is unknown: the selected paraffin-section caption does not report its fixative or fixation duration (datasheet A01967-2). Record each block’s fixative and processing history, and compare sections of similar thickness in the same staining run (standard IHC practice). For newly collected material, document the duration of fixation in 10% neutral buffered formalin if that is the laboratory’s chosen tissue workflow (standard IHC practice). Keep EDTA retrieval at pH 8.0 and the primary incubation constant while comparing blocks (datasheet A01967-2; standard IHC practice). Do not attribute a block difference to MFSD2A epitope sensitivity without a controlled fixation comparison (standard IHC practice).
Should MFSD2A stain cell borders, cytoplasm, or both in tissue IHC?
Prioritize a cell-border pattern when assessing MFSD2A, while allowing intracellular signal consistent with endoplasmic reticulum membrane and reported cytoplasmic punctae (UniProt Q8NA29 subcellular location). The protein has 12 transmembrane segments, so a diffuse nuclear pattern would not match its annotated topology (UniProt Q8NA29 topology). HPA reports mainly plasma-membrane localization, with cytosol marked uncertain and cytoplasmic bodies additionally observed in cell images (HPA subcellular). Its tissue-IHC profile also describes cytoplasmic expression across most tissues, but that dataset has uncertain reliability (HPA tissue IHC). Judge the compartment within identifiable cells and compare no-primary controls before treating granular DAB deposits as intracellular localization (standard IHC practice).
How should I assess an apparent epitope-dependent staining pattern?
Check the catalog antibody’s stated immunogen against the 3 annotated MFSD2A isoforms before interpreting a missing stain as absent protein (UniProt Q8NA29 isoforms; standard IHC practice). The supplied record does not map this antibody’s epitope or establish which isoforms it recognizes (datasheet A01967-2; UniProt Q8NA29 isoforms). MFSD2A contains 12 membrane-spanning segments, cytoplasmic regions, extracellular loops, and glycosylation sites at residues 230 and 240 (UniProt Q8NA29 topology and glycosylation). Those features make epitope position relevant to interpreting accessibility, but they do not identify this antibody’s binding site (UniProt Q8NA29 topology; standard IHC practice). Compare staining from an independently mapped antibody only after checking its tissue-IHC suitability and controls (standard IHC practice).
How can I assess MFSD2A by multiplex IF alongside this IHC guide?
For the separate IF workflow, pair MFSD2A with a marker of brain microvascular endothelium when assessing its blood-brain-barrier context (UniProt Q8NA29 function; standard IF practice). Select spectrally separated fluorophores and check unstained tissue autofluorescence, favoring a far-red channel for a weak target when that background is lower (standard IF practice). Check the antibody epitope before permeabilisation: an intracellular-facing epitope needs membrane access, whereas a surface-facing extracellular epitope can be examined without permeabilisation (UniProt Q8NA29 topology; standard IF practice). The supplied catalog caption documents paraffin-section chromogenic IHC, not an IF procedure for this antibody (datasheet A01967-2). Validate IF staining with single-label, no-primary, and compartment controls before comparing it with DAB results (standard IF practice).
How do I reduce diffuse DAB background without losing MFSD2A signal?
Begin with a no-primary section to separate detection-system deposits from primary-antibody-associated staining (standard IHC practice). The selected paraffin-section example used 10% goat serum, 2 μg/ml rabbit primary overnight at 4°C, a peroxidase-conjugated secondary for 30 minutes at 37°C, and DAB development (datasheet A01967-2). Add an endogenous-peroxidase block, thorough washes, and a controlled DAB development time as general chromogenic IHC steps (standard IHC practice). If background persists, titrate primary concentration downward while keeping retrieval at EDTA pH 8.0 initially (datasheet A01967-2; standard IHC practice). Evaluate tissue edges and necrotic areas separately from intact cells when judging improvement (standard IHC practice).
What should I quantify when MFSD2A staining is heterogeneous? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then record the percentage of positive cells and intensity on a 0–3 scale (standard IHC practice). An H-score sums each intensity category multiplied by its cell percentage, yielding a 0–300 range; report the scoring rules alongside results (standard IHC practice). For vessel-focused analysis, measure MFSD2A-positive microvessels per mm² of viable tissue and normalize against all evaluable microvessels in the same region (UniProt Q8NA29 function; standard IHC practice). Keep imaging, DAB development, and threshold settings consistent across sections (standard IHC practice). Report membrane-associated and cytoplasmic staining separately because both locations have supporting observations with different confidence levels (HPA subcellular; HPA tissue IHC).
What distinguishes credible MFSD2A staining from tissue artefact?
A credible pattern should occur in identifiable cells and accord with MFSD2A’s plasma-membrane and endoplasmic-reticulum-membrane annotations (UniProt Q8NA29 subcellular location; standard IHC practice). Brain microvascular endothelial staining has biological context, whereas HPA’s broad cytoplasmic tissue-IHC profile carries uncertain reliability (UniProt Q8NA29 function; HPA tissue IHC). Treat dominant nuclear staining, isolated tissue-edge deposits, and staining restricted to necrotic regions as reasons to investigate artefact rather than assign positivity (UniProt Q8NA29 topology; standard IHC practice). Compare intact regions with the no-primary section and verify that endogenous peroxidase was blocked before interpreting DAB signal (standard IHC practice). A negative result alone cannot establish absent MFSD2A without suitable tissue and staining controls (standard IHC practice).
Boster reagents
Best MFSD2A / Sodium-dependent lysophosphatidylcholine symporter 1 IHC Antibodies
A01967-2 has IHC images from paraffin-embedded human liver and liver cancer sections, plus IF images from SIHA cells and human liver cancer sections (catalog image captions).
Real IHC data
Anti-MFSD2A Antibody ®
Cat # A01967-2
A01967-2 is listed for IHC, IF and ICC, with Human, Mouse and Rat reactivity (catalog applications and reactivity). Its IHC images show paraffin-embedded human liver and liver cancer sections; its IF images show SIHA cells and a paraffin-embedded human liver cancer section (catalog image captions).
Which to pick: For tissue IHC, choose A01967-2: its own IHC captions document paraffin-embedded human sections at 2 μg/ml after heat retrieval in EDTA, pH 8.0; the fixative is unreported (catalog IHC captions). For IF/ICC, A01967-2 is also listed for both applications, with IF images of SIHA cells and human liver cancer tissue at 5 μg/ml (catalog applications and IF captions). For cross-species work, A01967-2 is a polyclonal antibody listed as reactive with Human, Mouse and Rat, although its supplied IHC and IF images show human samples only (catalog dilution record, reactivity and image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8NA29 (NLS1_HUMAN, Sodium-dependent lysophosphatidylcholine symporter 1).
- Human Protein Atlas. MFSD2A tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. MFSD2A subcellular location (ICC-IF): Mainly localized to the plasma membrane and cytosol. In addition localized to the cytoplasmic bodies..
- Human Protein Atlas. MFSD2A antibody validation summary (1 antibodies).
- MFSD2A is a novel lung tumor suppressor gene modulating cell cycle and matrix attachment. Molecular cancer 2010 — PMC2846890.
- Mfsd2a suppresses colorectal cancer progression and liver metastasis via the S100A14/STAT3 axis. Journal of translational medicine 2025 — PMC11726956.
- The prognostic value of major facilitator superfamily domain-containing protein 2A in patients with hepatocellular carcinoma. Aging 2019 — PMC6814593.
- Mfsd2a is critical for the formation and function of the blood-brain barrier. Nature 2014 — PMC4134871.
- PubMed PMID:12975309 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15221005 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MFSD2A (Sodium-dependent lysophosphatidylcholine symporter 1). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC); membrane and ER localisation (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A01967-2). Validated IHC antibody: A01967-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative Heart muscle. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A01967-2); 2. Fixation: Image fixative and duration unreported (datasheet A01967-2); 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 A01967-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01967-2); 8. Primary antibody: Rabbit anti-MFSD2A, 2-5 μg/ml (datasheet A01967-2); 9. Primary incubation: Overnight at 4 °C (datasheet A01967-2). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining has low consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 3 isoforms; check extracellular versus cytoplasmic epitope (UniProt); Regulation: Tissue-enhanced RNA expression (HPA tissue RNA). Evidence: UniProt Q8NA29, HPA tissue IHC · Uncertain, HPA ICC-IF · Mainly localized to the plasma membrane and cytosol. In addition…, Datasheet A01967-2, PMC2846890, PMC6814593. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MFSD2A/immunohistochemistry-ihc-guide.