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MARCKS / Myristoylated alanine-rich C-kinase substrate · IHC design guide
Design Immunohistochemistry for MARCKS
Plan MARCKS chromogenic IHC in paraffin sections using appendix glandular cells or kidney tubules as high-staining reference tissues (HPA tissue IHC). Interpret cytoplasmic and membranous staining with the IHC-validated antibody, while allowing for PKC-dependent redistribution between those compartments (HPA tissue IHC; UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MARCKS 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 and membranous staining (HPA tissue IHC) | |
| Staining pattern | Glandular and respiratory epithelial cells: cytoplasmic/membranous (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A04133-4) | |
| 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 paraffin-section fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A04133-4) | |
| Caveat | Tissue staining has medium RNA concordance (HPA tissue IHC) | |
| Regulation | PKC phosphorylation shifts membrane signal (UniProt) | |
| Isoform / epitope | No isoforms annotated; epitope map unspecified (UniProt) |
Section 1
Recommended MARCKS IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet A04133-4) with two published MARCKS IHC protocols (PMC5351628; PMC5795991).
| Sample | Paraffin-embedded mouse brain tissue; fixative not specified (datasheet A04133-4) |
| Fixation | Image fixative and duration unreported (datasheet A04133-4); 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 A04133-4); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A04133-4) |
| Primary antibody | Rabbit anti-MARCKS, 2-5 μg/ml (datasheet A04133-4) |
| Primary incubation | Overnight at 4 °C (datasheet A04133-4) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A04133-4) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | MARCKS-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic and membranous expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet A04133-4). Use PT Link at pH 6 when reproducing either published protocol (PMC5351628; PMC5795991).
Section 2
What Is the Expected MARCKS Staining Pattern?
In paraffin-section IHC, expect cytoplasmic and membranous MARCKS staining in many tissues, including glandular and respiratory epithelial cells, kidney tubule cells, and cerebellar neuropil (HPA tissue IHC: high in these cells or regions). MARCKS has no transmembrane segment and can move between membrane and cytoplasm (UniProt P29966: topology and subcellular location). HPA rates the tissue pattern Enhanced, with medium consistency between staining and RNA data pending external verification (HPA tissue IHC: reliability).
| Membranous and cytoplasmic staining marks glandular cells in colon or appendix, respiratory epithelium in bronchus, or kidney tubule cells; the cerebellar molecular-layer neuropil also stains (HPA tissue IHC: High). | These are supported positive patterns for a total-MARCKS stain. Score the specified cells or region and record membrane and cytoplasmic signal separately when distinguishable (HPA tissue IHC: cell-level pattern; UniProt P29966: subcellular location). HPA reports staining levels, not an expected staining percentage or a numerical scoring cutoff (HPA tissue IHC: High). |
| The dominant signal is nuclear with little membrane or cytoplasmic staining in an HPA-positive tissue (HPA tissue IHC: cytoplasmic and membranous profile). | Treat a nuclear-only IHC result as discordant and review the antibody, controls, and detection conditions (HPA tissue IHC: profile; standard IHC practice). HPA lists nucleoplasmic localisation as uncertain in ICC-IF, so that observation alone does not validate dominant nuclear staining in paraffin sections (HPA subcellular: nucleoplasm uncertain). |
| Cardiomyocytes, skeletal myocytes, or adipocytes stain strongly while the expected positive cells are weak (HPA tissue IHC: Not detected in those cell types; High in listed positive cells). | This distribution raises concern for nonspecific binding, cross-reactivity, or endogenous chromogenic activity; it does not identify which cause applies (standard IHC practice). Compare tissue architecture and matched controls. HPA's Not detected calls apply to the named cell types, not every cell in their tissues (HPA tissue IHC: cell-level negatives). |
| Brown signal spreads evenly through the section or appears outside recognizable cell boundaries (standard chromogenic IHC observation). | Diffuse deposition obscures the membrane and cytoplasmic pattern used to interpret MARCKS (HPA tissue IHC: profile). Check background on a no-primary control, then assess blocking, antibody concentration, washes, and chromogen development as general IHC variables (standard IHC practice). |
| A run shows no convincing signal in colon glandular cells, bronchial respiratory epithelium, or kidney tubule cells (HPA tissue IHC: High in these cells). | The result is inconclusive until a known-positive control works in the same run (standard IHC practice). Check the catalog antibody's IHC-P instructions, retrieval conditions, and detection reagents before calling the test tissue negative (standard IHC practice). The supplied sources do not establish MARCKS-specific fixation sensitivity. |
💡Expected MARCKS appearanceCall a positive result when recognizable glandular, respiratory epithelial, or kidney tubule cells show cytoplasmic and/or membranous staining, or cerebellar molecular-layer neuropil stains, consistent with HPA's High examples; diffuse deposit or dominant nuclear-only staining is a warning pattern (HPA tissue IHC: profile and levels; HPA subcellular: nucleoplasm uncertain).
| Compartment and phosphorylation state (UniProt P29966: subcellular location) | MARCKS associates with the cell membrane and cytoplasm; PKC-dependent phosphorylation displaces it from the membrane, and dephosphorylation accompanies reassociation (UniProt P29966: subcellular location). Therefore, relative membrane and cytoplasmic prominence can vary without changing the expected set of compartments. A total-MARCKS IHC image alone cannot establish phosphorylation state (UniProt P29966: phosphorylation-linked movement; standard interpretation). |
| Cell-level tissue reference (HPA tissue IHC: profile and examples) | HPA reports High staining in selected glandular cells, respiratory epithelial cells, kidney tubule cells, and cerebellar neuropil, alongside Not detected calls for specified adipocytes, cardiomyocytes, and myocytes (HPA tissue IHC: levels). Use the named cells as reference points; low tissue RNA specificity does not mean every cell should stain alike (HPA tissue IHC: RNA specificity and cell-level staining). |
| Strength of reference evidence (HPA tissue IHC: reliability; HPA antibodies: IHC validation) | The tissue profile is rated Enhanced, although its staining-to-RNA consistency is medium and external verification is pending (HPA tissue IHC: reliability description). HPA lists three antibodies with Enhanced IHC status (HPA antibodies: HPA067595, HPA069443, CAB022062). These observations support comparison with the pattern; they do not validate an unlisted catalog antibody or define a universal staining cutoff (HPA tissue IHC; standard interpretation). |
| Retrieval and epitope information (supplied UniProt P29966 and HPA records) | No epitope location or MARCKS-specific retrieval response is supplied. Follow the catalog antibody's IHC-P procedure and evaluate its stated retrieval condition with a positive control (standard IHC practice). The lack of a transmembrane segment does not identify an optimal retrieval method or predict fixation effects (UniProt P29966: topology). |
| Detection system background (standard chromogenic IHC practice) | Endogenous enzyme activity and nonspecific reagent binding can produce chromogenic signal unrelated to antibody binding (standard IHC practice). A no-primary control helps reveal detection-system background in the tested tissue, while a positive control checks that the run can reveal the expected cellular pattern (standard IHC practice; HPA tissue IHC: positive cell examples). |
| IF/ICC Q&A: where should fluorescence appear? (HPA subcellular: ICC-IF) | Mainly at the plasma membrane, which HPA marks supported; HPA also lists cytosol, nucleoplasm, and primary cilium as uncertain additional locations (HPA subcellular: locations). This ICC-IF observation helps interpret fluorescence images. It is separate from the paraffin-section IHC pattern and supplies no IF/ICC protocol option (HPA subcellular; HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in a section expected to contain colon glandular cells, bronchial respiratory epithelium, or kidney tubules (HPA tissue IHC: High in these cells). | The run may have failed at antibody incubation, antigen retrieval, or detection; the absent signal alone cannot identify the failed step (standard IHC practice). | Run a known-positive section alongside the sample, confirm tissue preservation and the catalog antibody's IHC-P instructions, and check detection reagents (standard IHC practice). Interpret a negative sample only after the positive control stains in the expected cells (HPA tissue IHC: High examples). |
| The entire section develops diffuse brown colour, including areas without clear cellular staining (standard chromogenic IHC observation). | Possible causes include detection background, insufficient blocking or washing, excessive primary-antibody concentration, or prolonged chromogen development (standard IHC practice). | Review a no-primary control and reagent controls; then adjust blocking, washes, antibody concentration, or development time one variable at a time (standard IHC practice). Reassess whether membrane and cytoplasmic boundaries are readable (HPA tissue IHC: expected profile). |
| A strong nuclear-only pattern replaces membrane and cytoplasmic staining in otherwise positive cells (HPA tissue IHC: expected profile). | The pattern is discordant with tissue IHC; HPA's additional nucleoplasmic ICC-IF location is uncertain and does not explain a nuclear-only paraffin result by itself (HPA subcellular: nucleoplasm uncertain; HPA tissue IHC: profile). | Compare with a no-primary control, verify antibody and detection setup, and examine an HPA-positive tissue before assigning biological meaning (standard IHC practice; HPA tissue IHC: High examples). |
| Adipocytes, cardiomyocytes, or skeletal myocytes appear strongly positive (HPA tissue IHC: Not detected in these named cell types). | Nonspecific staining or endogenous chromogenic activity is possible; HPA's cell-level negatives cannot determine the cause from one slide (standard IHC practice; HPA tissue IHC: negatives). | Confirm which cells carry the colour, inspect a no-primary control, and compare a named positive cell population in the same run (standard IHC practice; HPA tissue IHC: positives and negatives). |
| Membrane staining is weaker than cytoplasmic staining, but the expected cells retain a readable signal (HPA tissue IHC: cytoplasmic and membranous profile). | MARCKS can shift between membrane and cytoplasm with phosphorylation and dephosphorylation; relative intensity does not establish that mechanism in this specimen (UniProt P29966: phosphorylation-linked movement). | Record both compartments and score the named cells against controls using the same readout criteria (standard IHC practice; HPA tissue IHC: cell-level pattern). Do not infer phosphorylation status from total-MARCKS chromogenic staining alone (UniProt P29966: subcellular movement). |
| An IF/ICC image shows fluorescence outside the plasma membrane (HPA subcellular: ICC-IF locations). | HPA calls plasma membrane localisation supported but labels cytosol, nucleoplasm, and primary cilium uncertain; an additional location is therefore not independently confirmed by that listing (HPA subcellular: location confidence). | Interpret the image within the separate IF/ICC guide using its own controls, and avoid transferring an uncertain ICC-IF location into the paraffin-section IHC scoring rule (HPA subcellular: ICC-IF; HPA tissue IHC: profile; standard interpretation). |
Experiment evidence
IHC & IF images showing MARCKS staining
A04133-4 · IHCIHC analysis of MARCKS using anti-MARCKS antibody (A04133-4). MARCKS was detected in a paraffin-embedded section of mouse brain tissue. Heat
A04133-4 · IHCIHC analysis of MARCKS using anti-MARCKS antibody (A04133-4). MARCKS was detected in a paraffin-embedded section of rat brain tissue. Heat m
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000277443-MARCKS/tissue/cerebral+cortex#imid_19572279
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000277443-MARCKS/tissue/colon#imid_19572205
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000277443-MARCKS/tissue/liver#imid_19572198
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000277443-MARCKS/tissue/kidney#imid_19572199
HPA · ICC-IF SH-SY5YHuman Protein Atlas immunofluorescence, SH-SY5Y, antibody HPA067595; green = MARCKS, blue = nucleus, red = microtubules.
HPA · ICC-IF SH-SY5YHuman Protein Atlas immunofluorescence, SH-SY5Y, antibody HPA067595; green = MARCKS, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MARCKS IHC & IF
🧪Run appendix first and require staining in its glandular cells (HPA: High in appendix glandular cells). Use adipose tissue adipocytes as the negative tissue (HPA: Not detected in adipocytes); on the appendix slide, assess cells outside the glandular compartment as an internal background reference without assuming every such cell is negative.
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MARCKS in SH-SY5Y, U-251MG, SiHa, SuSa, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), KOLF2.1J, with annotated localisation: Plasma membrane (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and an isotype-matched rabbit IgG control appropriate to the primary antibody (selected-SKU caption: rabbit anti-MARCKS). Confirm specificity with MARCKS-knockout material or a validated peptide-block control; for appendix DAB staining, quench endogenous peroxidase and inspect inflammatory cells for background (selected-SKU caption: peroxidase detection with DAB).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected-SKU paraffin-section caption does not state the fixative (selected-SKU caption: fixative unreported). The demonstrated paraffin-section method uses heat retrieval in EDTA at pH 8.0, but the evidence does not establish that retrieval is required or that frozen sections or IF are easier (selected-SKU caption: EDTA heat retrieval). In appendix, glandular architecture and possible peroxidase-positive inflammatory cells warrant compartment-specific scoring and background checks; membrane-associated and cytoplasmic signal are biologically plausible (HPA: High in appendix glandular cells; UniProt P29966: cell membrane and cytoplasm).
HPA tissue IHC evidence for MARCKS
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium 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 → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Molecular layer - neuropil | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | 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 → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MARCKS IHC Tips
Use the catalog antibody’s paraffin section protocol as the starting point, then assess MARCKS staining by cell type, compartment and controls.
Which retrieval condition should I start with for MARCKS paraffin sections?
Start with heat mediated retrieval in EDTA at pH 8.0 (datasheet A04133-4). The catalog antibody’s mouse brain paraffin section example used this condition, followed by 10% goat serum blocking and 2 μg/ml primary antibody overnight at 4°C (datasheet A04133-4). If staining is weak, compare a shorter and longer heating cycle on matched sections while holding antibody concentration and detection constant (standard IHC practice). Judge the result by preserved tissue morphology and expected cytoplasmic or membranous staining, rather than by overall DAB intensity alone (HPA tissue IHC profile; standard IHC practice).
Could fixation explain weak or patchy MARCKS staining?
The selected image identifies a paraffin embedded mouse brain section but does not state its fixative, so target specific fixation sensitivity is unknown (datasheet A04133-4). Record the fixative, fixation duration, tissue thickness and processing history for each specimen before comparing staining across batches (standard IHC practice). On matched sections, first apply the documented EDTA pH 8.0 retrieval and 2 μg/ml primary antibody conditions consistently (datasheet A04133-4). If signal differs between processing batches, assess morphology and control tissue in parallel; that comparison can identify a processing association without establishing a MARCKS specific fixation effect (standard IHC practice).
Should MARCKS stain cell borders or cytoplasm in chromogenic IHC?
Both membranous and cytoplasmic staining are plausible: MARCKS associates with the cell membrane and cytoskeleton, and tissue IHC shows both patterns across many tissues (UniProt P29966 subcellular; HPA tissue IHC profile). PKC dependent phosphorylation can displace MARCKS from the membrane, with dephosphorylation accompanying reassociation, so compartment patterns may vary with cell state (UniProt P29966 subcellular). Score border and cytoplasmic DAB staining separately within the same defined cell population, using a counterstain to identify cells and tissue structure (standard IHC practice). Do not assign isolated nuclear DAB as expected tissue staining solely because an uncertain nucleoplasmic IF location is listed (HPA subcellular; standard IHC practice).
How do epitope uncertainty and MARCKS phosphorylation affect staining interpretation?
The supplied record lists 0 annotated isoforms and multiple modified residues, including phosphoserines at positions 26, 27 and 29 (UniProt P29966 isoforms; UniProt P29966 modified residues). No antibody epitope or phosphorylation dependence is supplied, so a change in DAB signal cannot be assigned to one modification (datasheet A04133-4; UniProt P29966 modified residues). Keep retrieval at EDTA pH 8.0 and antibody at 2 μg/ml when comparing specimens, since both conditions are documented for the catalog image (datasheet A04133-4). If the biological question concerns phosphorylation, assess it with an independently characterized phospho specific reagent or orthogonal assay (standard IHC practice).
How can IF help check an ambiguous MARCKS IHC pattern?
Use IF as a separate check of cell identity and compartment when an IHC DAB pattern is ambiguous (standard IF practice). Multiplex MARCKS with a validated marker for the cell population under study; for example, a neuronal marker can help interpret staining in the mouse brain setting shown by the catalog image (datasheet A04133-4; standard IF practice). Choose fluorophores after checking tissue autofluorescence and include single stain controls to distinguish bleed through from colocalisation (standard IF practice). Because MARCKS has no transmembrane segment and can occupy the cytoplasmic face of the membrane, test permeabilisation for intracellular epitope access; the antibody’s epitope side is unreported (UniProt P29966 topology and subcellular; datasheet A04133-4).
What should I change when MARCKS DAB staining is diffuse?
Compare the stained section with a no primary control and inspect whether pigment persists without the catalog antibody (standard IHC practice). The catalog example used 10% goat serum blocking, a peroxidase linked secondary and DAB development, so verify blocking and detection steps when diffuse signal appears (datasheet A04133-4). Apply an endogenous peroxidase block and check reagent only controls where tissue enzyme activity could contribute DAB signal (standard chromogenic IHC practice). Titrate primary antibody around the documented 2 μg/ml condition on matched sections, retaining expected membranous or cytoplasmic localisation and tissue morphology as decision criteria (datasheet A04133-4; HPA tissue IHC profile; standard IHC practice).
How should I quantify MARCKS staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue region before scoring, then apply the same DAB exposure and image analysis settings to every section (standard IHC practice). For cellular staining, report the percentage of positive cells and an H score combining intensity categories with their cell percentages; score membranous and cytoplasmic signal separately (standard IHC practice; HPA tissue IHC profile). Where individual cells cannot be resolved reliably, report positive area or integrated DAB density per mm² of viable tissue instead (standard IHC practice). Normalize counts to eligible cells and area based measurements to viable tissue area, and compare matched controls within each staining batch (standard IHC practice).
How can I distinguish genuine MARCKS signal from section artefact?
Expect interpretable MARCKS signal in cytoplasmic or membranous compartments, consistent with its membrane association and the tissue IHC profile (UniProt P29966 subcellular; HPA tissue IHC profile). Check the stained cell type: HPA reports high staining in kidney tubule cells and no detected staining in adipocytes, although its tissue IHC reliability remains pending external verification (HPA tissue IHC). Treat staining confined to section edges, necrotic regions or disrupted tissue as suspect, and compare it with intact regions on the same slide (standard IHC practice). A no primary control helps identify detection background, while an endogenous peroxidase control helps assess enzyme derived DAB deposits (standard chromogenic IHC practice).
Boster reagents
Best MARCKS / Myristoylated alanine-rich C-kinase substrate IHC Antibodies
The catalog antibody has real paraffin-section IHC images from mouse and rat brain and human stomach cancer tissue (A04133-4 image captions); no IF image is supplied (catalog payload).
Real IHC data
Anti-MARCKS Antibody ®
Cat # A04133-4
A04133-4 is listed for human, mouse and rat IHC, with paraffin-section images from human stomach cancer tissue and mouse and rat brain (catalog applications/reactivity; A04133-4 image captions). A04133S163 is listed for human, mouse and rat IHC targeting phospho-S163, but has no IHC image in the payload (catalog applications/reactivity; A04133S163 catalog entry).
Which to pick: Choose A04133-4 for tissue IHC and cross-species comparisons: its own captions document paraffin sections from all three listed species, while the fixative and clonality are unreported (A04133-4 image captions; catalog reactivity/clone field). Choose A04133S163 when phospho-S163 is the target, allowing for its lack of an IHC image; its listed IHC dilution is 1:50–1:200 and clonality is unreported (A04133S163 catalog entry). Neither SKU lists IF/ICC validation or supplies an IF image, so there is no catalog-supported IF/ICC pick here (catalog applications; IF image fields).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P29966 (MARCS_HUMAN, Myristoylated alanine-rich C-kinase substrate).
- Human Protein Atlas. MARCKS tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. MARCKS subcellular location (ICC-IF): Mainly localized to the plasma membrane. In addition localized to the nucleoplasm, cytosol and primary cilium..
- Human Protein Atlas. MARCKS antibody validation summary (3 antibodies).
- MARCKS protein overexpression in inflammatory breast cancer. Oncotarget 2017 — PMC5351628.
- Stromal Expression of MARCKS Protein in Ovarian Carcinomas Has Unfavorable Prognostic Value. International journal of molecular sciences 2017 — PMC5795991.
- Upregulation of MARCKS in kidney cancer and its potential as a therapeutic target. Oncogene 2017 — PMC5926797.
- MARCKS contributes to stromal cancer-associated fibroblast activation and facilitates ovarian cancer metastasis. Oncotarget 2016 — PMC5122339.
- PubMed PMID:1860846 — UniProt-cited evidence.
- PubMed PMID:1427823 — UniProt-cited evidence.
- PubMed PMID:14574404 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MARCKS (Myristoylated alanine-rich C-kinase substrate). Expected localisation Cytoplasmic and membranous staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A04133-4). Validated IHC antibody: A04133-4 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse brain tissue; fixative not specified (datasheet A04133-4); 2. Fixation: Image fixative and duration unreported (datasheet A04133-4); 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 A04133-4); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A04133-4); 8. Primary antibody: Rabbit anti-MARCKS, 2-5 μg/ml (datasheet A04133-4); 9. Primary incubation: Overnight at 4 °C (datasheet A04133-4). Watch for: Fixation: Keep paraffin-section fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A04133-4); Caveat: Tissue staining has medium RNA concordance (HPA tissue IHC); Isoform / epitope: No isoforms annotated; epitope map unspecified (UniProt); Regulation: PKC phosphorylation shifts membrane signal (UniProt). Evidence: UniProt P29966, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the plasma membrane. In addition localized to…, Datasheet A04133-4, PMC5351628, PMC5795991. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MARCKS/immunohistochemistry-ihc-guide.