This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
MCM5 / DNA replication licensing factor MCM5 · IHC design guide
Design Immunohistochemistry for MCM5
Plan chromogenic IHC for MCM5 in paraffin sections using the catalog antibody's documented protocol (datasheet M03642-2). Assess nuclear staining in the cell types reported by tissue IHC, including hematopoietic and immune cells (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MCM5 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 | Nuclear staining in selected tissue cells (HPA tissue IHC) | |
| Staining pattern | Nuclear staining, especially in hematopoietic and immune cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M03642-2) | |
| 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) | |
| Caveat | Chromatin association changes during DNA replication (UniProt) | |
| Regulation | Expression regulation is not annotated (UniProt) | |
| Isoform / epitope | No annotated isoforms; the chain spans residues 2–734 (UniProt) |
Section 1
Recommended MCM5 IHC & IF Protocols
The catalog antibody uses heat-mediated EDTA retrieval (datasheet M03642-2). The three published IHC protocols below report staining of oral tissue, tissue sections, and lung tissue or mouse tumors (PMC10829438; PMC2391096; PMC8366603).
| Sample | Paraffin-embedded human gallbladder adenocarcinoma tissue; fixative not specified (datasheet M03642-2) |
| Fixation | Image fixative and duration unreported (datasheet M03642-2); 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 M03642-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M03642-2) |
| Primary antibody | Mouse monoclonal (clone 4G10) anti-MCM5, 2μg/ml (datasheet M03642-2) |
| Primary incubation | Overnight at 4 °C (datasheet M03642-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet M03642-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | MCM5-positive staining in germinal center cells of appendix (HPA tissue IHC: High). HPA tissue profile: Nuclear expression mainly in hematopoietic cells, immune cells, cytotrophoblastic cells in placenta and cells in testis. Also expressed in other cell types. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet M03642-2). Microwave citrate retrieval is a published alternative for the lung study (PMC8366603).
Section 2
What Is the Expected MCM5 Staining Pattern?
MCM5 should appear predominantly in nuclei, with chromatin association that changes as DNA replication progresses (UniProt P33992). In tissue IHC, expect prominent staining in hematopoietic and immune cells, placental trophoblastic cells, and selected other cell types (HPA tissue IHC; reliability: Enhanced). MCM5 has no transmembrane segment, so a membrane outline is inconsistent with its annotated location (UniProt P33992 topology).
| Distinct nuclear staining in bone marrow hematopoietic cells or lymph node germinal center cells, with limited surrounding background. | This matches HPA's High IHC staining in those cell populations (HPA tissue IHC). Judge localization within the identified cells; a whole-section average can obscure a strong signal confined to a particular population. |
| Predominantly cytoplasmic or membrane staining, especially when nuclei are clear. | The compartment conflicts with nuclear and chromosome localization (UniProt P33992) and HPA's nuclear tissue profile (HPA tissue IHC). Treat it as a possible staining artefact; review morphology and detection controls before assigning it to MCM5. |
| Strong staining in adipocytes or cardiomyocytes, while the expected positive cells stain appropriately. | HPA reports MCM5 as Not detected in those cell types (HPA tissue IHC). This discordance raises possible antibody cross-reactivity or endogenous detection activity; it does not, by itself, identify which is responsible. |
| Haze or granular color across nuclei, cytoplasm, and extracellular areas, with little distinction between cell populations. | The distribution is difficult to reconcile with the expected nuclear pattern (UniProt P33992; HPA tissue IHC). Evaluate it as nonspecific background using a no-primary control and the spatial pattern of staining before scoring positive nuclei. |
| No convincing nuclear signal in bone marrow hematopoietic cells or appendix germinal center cells. | Both are reported High by HPA (HPA tissue IHC), so an entirely blank known-positive region warrants a technical review. First confirm that the relevant cells are present; a negative field without them is not an informative control. |
💡Expected MCM5 appearanceCall a convincing positive result when identifiable bone marrow hematopoietic cells or germinal center cells show clear, often strong nuclear staining (HPA: High); dominant membrane, cytoplasmic, or diffuse extracellular color is discordant with MCM5 localization (UniProt P33992; HPA tissue IHC).
| Cell population and tissue context | HPA reports High staining in skin basal-layer cells, placental trophoblastic cells, duodenal Paneth cells, and small-intestinal enterocytes (HPA tissue IHC). Compare like cell types; adjacent cells may have different staining levels. |
| Low and undetected reference populations | Lung macrophages and stomach glandular cells are Low, whereas adipocytes and cardiomyocytes are Not detected (HPA tissue IHC). Low staining is a weaker benchmark than a High population; Not detected is an observed IHC category, not proof of biological absence. |
| Compartment during replication | MCM5 is nuclear and chromosome associated; its chromatin association changes as DNA replication progresses (UniProt P33992). Variation in nuclear appearance can therefore be assessed in context, while predominant membrane outlining remains discordant with its annotated topology. |
| Antibody and application validation | HPA lists Enhanced IHC validation for HPA000845 and CAB000101, and Enhanced ICC validation for HPA052880 (HPA antibodies). These statuses support the stated applications for those antibodies; they do not establish interchangeable performance across IHC and ICC. |
| Target-specific retrieval evidence | Target-specific fixation effects are not established by the supplied assay evidence. Verify with a matched IHC source before attributing a result to fixation. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive tissue has no nuclear signal. | The sampled field may lack the expected cells, or an IHC workflow step may have failed; HPA reports High staining in bone marrow hematopoietic cells (HPA tissue IHC). | Confirm the cell population on the counterstained section. Run a known-positive section alongside the sample, then check retrieval, primary-antibody application, and detection steps as general IHC workflow checks. |
| Color is strongest at cell membranes or in cytoplasm. | This conflicts with nuclear and chromosome localization and the absence of a transmembrane segment (UniProt P33992). It may reflect nonspecific binding or detection artefact. | Compare the same compartments in a no-primary control, check that nuclear morphology is preserved, and score only convincing nuclear staining in the expected cell population. |
| Adipocytes or cardiomyocytes stain strongly. | HPA reports these cell populations as Not detected (HPA tissue IHC); cross-reactivity or endogenous detection activity is possible, but the image alone cannot distinguish them. | Review a no-primary control for detection-derived color, then compare the candidate signal with a High reference population on the same run (HPA tissue IHC). |
| Diffuse color obscures the difference between positive and negative cells. | Broad background can arise from nonspecific binding or the chromogenic detection workflow; it does not resemble the reported nuclear tissue pattern (HPA tissue IHC). | Review the no-primary control, blocking and wash steps, and detection exposure. Reassess whether nuclear signal remains distinct in HPA High cells (HPA tissue IHC). |
| Only a few cells stain in a region expected to be positive. | Tissue labels do not imply uniform staining: HPA identifies specific High populations, including skin basal-layer cells and appendix germinal center cells (HPA tissue IHC). | Locate and score the named population before judging the run. If those cells are present but unstained, compare them with a concurrent known-positive section and review the IHC workflow. |
| IF/ICC Q&A: What localization should be checked? | HPA reports enhanced nucleoplasmic localization by ICC-IF, while UniProt annotates MCM5 in the nucleus and on chromosomes (HPA subcellular; UniProt P33992). | Check whether fluorescence overlaps the nuclear counterstain and is concentrated in the nucleoplasm (HPA subcellular). Use the separate IF/ICC guide for its workflow; IHC antibody validation does not establish ICC performance (HPA antibodies). |
Experiment evidence
IHC & IF images showing MCM5 staining
M03642-2 · IHCIHC analysis of MCM5 using anti-MCM5 antibody (M03642-2). MCM5 was detected in paraffin-embedded section of human gallbladder adenocarcinoma
M03642-2 · IHCIHC analysis of MCM5 using anti-MCM5 antibody (M03642-2). MCM5 was detected in paraffin-embedded section of human lung cancer tissue. Heat m
M03642-2 · IFIF analysis of MCM5 using anti-MCM5 antibody (M03642-2). MCM5 was detected in immunocytochemical section of A431 cells. Enzyme antigen retri
PA1653 · IHCAnti-MCM5 antibody, PA1653, IHC(P) IHC(P): Human Intestinal Cancer Tissue
PA1653 · IHCAnti-MCM5 antibody, PA1653, IHC(P) IHC(P): Human Intestinal Cancer Tissue
PA1653 · IFIF analysis of MCM5 using anti-MCM5 antibody (PA1653). MCM5 was detected in immunocytochemical section of U20S cells. Enzyme antigen retriev
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000100297-MCM5/tissue/cerebral+cortex#imid_259486
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000100297-MCM5/tissue/colon#imid_259396
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000100297-MCM5/tissue/liver#imid_259413
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000100297-MCM5/tissue/kidney#imid_259406
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA052880; green = MCM5, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA052880; green = MCM5, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MCM5 IHC & IF
🧪Run lymph node first: germinal center cells should stain (HPA: High in lymph node germinal center cells); use adipose tissue adipocytes as the negative tissue (HPA: Not detected in adipocytes). On the positive slide, nuclei of adjacent cells without detectable MCM5 should lack chromogen; assess these as internal comparators without assuming every surrounding cell is negative.
Positive control tissue: Appendix (Germinal center cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MCM5 in A-431, U-251MG, U2OS, U2OS, siRNA 1 (10x), U2OS, siRNA 1 (40x), U2OS, siRNA 2 (10x), U2OS, siRNA 2 (40x), U2OS, scrambled (10x), U2OS, scrambled (40x), NIH 3T3, with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide, a concentration-matched mouse IgG isotype control (selected-SKU caption: mouse anti-MCM5 antibody), and identically processed MCM5-knockout material if available. For chromogenic lymph node IHC, block endogenous peroxidase and check for background in phagocytes (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported in the supplied evidence; the selected-SKU paraffin-section caption does not state the fixative. Heat retrieval in EDTA at pH 8.0 is a documented IHC starting condition (selected-SKU caption), but retrieval dependence in lymph node is unreported. ICC-IF images exist for U2OS (HPA: subcellular), while the supplied evidence does not establish whether IF or frozen sections are easier than paraffin IHC; check lymph node sections for endogenous-peroxidase background (standard IHC practice).
HPA tissue IHC evidence for MCM5
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High 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 |
|---|---|---|---|---|
| Appendix | Germinal center cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Duodenum | Paneth cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | High | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MCM5 IHC Tips
Troubleshoot MCM5 chromogenic IHC by checking nuclear staining, tissue controls, and the retrieval conditions documented for the catalog antibody.
What retrieval should I try when MCM5 nuclear staining is weak?
Start with heat-mediated EDTA at pH 8.0 for this paraffin-section assay (datasheet M03642-2). The selected tissue image used that retrieval before overnight incubation with 2 μg/ml antibody at 4°C (caption M03642-2). If nuclei remain weak, check that sections were fully deparaffinised and heated consistently across the slide before changing retrieval conditions (standard IHC practice). Compare any adjusted retrieval against a concurrently processed positive control, such as lymph-node germinal center cells, which show high expression (HPA: High in lymph-node germinal center cells). Excess retrieval can damage morphology, so judge signal alongside intact nuclear detail (standard IHC practice).
Could fixation explain weak or uneven MCM5 staining?
Target-specific sensitivity to fixation is unknown because the selected paraffin-section caption does not report its fixative (caption M03642-2). Record the fixative, fixation duration, tissue thickness, and processing history for each specimen before attributing differences to MCM5 biology (standard IHC practice). Compare sections processed together and inspect morphology: poorly preserved or unevenly fixed areas can make nuclear scoring unreliable (standard IHC practice). Use the documented EDTA pH 8.0 heat retrieval as the initial condition, then assess any changes with the same control tissue and detection settings (datasheet M03642-2; standard IHC practice). Do not infer a fixation effect from tissue-expression patterns or MCM5 modifications (HPA tissue IHC; UniProt P33992).
How should I investigate MCM5 signal outside nuclei?
Expect predominantly nuclear chromogenic staining: MCM5 is annotated in the nucleus and on chromosomes, while the subcellular atlas places it in the nucleoplasm (UniProt P33992; HPA subcellular). Confirm that the DAB deposit lies within counterstained nuclei at high magnification, especially where cells overlap (standard IHC practice). Widespread cytoplasmic or extracellular colour warrants review of the no-primary control, detection background, and tissue integrity before it is scored as MCM5 (standard IHC practice). MCM5 has no annotated transmembrane segment, so a crisp membrane pattern would be unexpected (UniProt P33992 topology). Chromatin association changes during replication, so nuclear intensity need not be identical in every cell (UniProt P33992 subcellular annotation).
Can epitope location explain patchy MCM5 staining?
The supplied record lists no MCM5 isoforms and places its AAA(+) ATPase domain at residues 314–654 (UniProt P33992). It also lists modifications including phosphoserine at 315 and 605 and acetyllysine at 392 and 396 (UniProt P33992). The antibody epitope is not supplied, so these sites cannot establish why a section stains weakly or patchily (supplied M03642-2 caption; UniProt P33992). First compare nuclear signal in matched positive controls under the same retrieval and antibody conditions (standard IHC practice). If performance differs between antibody lots, request epitope information and validate the new lot on the same tissue panel before interpreting biological differences (standard IHC practice).
How can I assess MCM5 by IF alongside a cell-type marker?
For IF/ICC, design the multiplex panel around nuclear MCM5 and a marker identifying the cell population being examined (UniProt P33992 localisation; standard IF practice). The subcellular atlas reports nucleoplasmic MCM5 and includes ICC/IF images, but those images do not establish an IF protocol for catalog antibody M03642-2 (HPA subcellular; caption M03642-2). Choose spectrally separated fluorophores after checking tissue autofluorescence, and include single-stain controls to assess bleed-through (standard IF practice). Because MCM5 has no transmembrane segment and is nuclear, permeabilisation must permit antibody access to the nucleus; optimise it while preserving nuclear morphology (UniProt P33992 topology and localisation; standard IF practice). Score colocalisation in identified cells rather than from colour overlap alone (standard IF practice).
What should I check when DAB obscures MCM5-positive nuclei?
First distinguish diffuse staining from discrete nuclear DAB signal using a no-primary control and the same development conditions (standard IHC practice). The selected image used 10% goat-serum blocking, 2 μg/ml primary antibody overnight at 4°C, and a biotin-based detection sequence (caption M03642-2). Check endogenous peroxidase activity and, with biotin-based detection, endogenous biotin when background follows tissue structures instead of nuclei (standard IHC practice). Shorten chromogen development or adjust antibody concentration only while retaining a concurrently stained positive control (standard IHC practice). MCM5 is expected in nuclei, so broad stromal or extracellular colour should not be counted as positive cells (UniProt P33992 localisation; standard IHC practice).
How should I quantify MCM5 IHC across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue region before scoring, then report the percentage of nuclei judged positive and the denominator of evaluable nuclei (standard IHC practice). An H-score can combine the percentages at intensity grades 0–3 into a 0–300 summary, provided thresholds are fixed across slides (standard IHC practice). For spatial comparisons, report positive-nucleus density per mm² of viable tissue and also the total nuclear density (standard IHC practice). Normalise comparisons to the same cell type, sampled area, staining run, and control performance rather than mixing populations with different expected expression (standard IHC practice; HPA tissue IHC). Exclude folds, necrosis, and poorly preserved regions before calculating scores (standard IHC practice).
When is an apparent MCM5-positive cell likely to be an artefact?
Treat a signal as credible when DAB is confined to intact nuclei in an appropriate cell population and the run controls perform as expected (UniProt P33992 localisation; standard IHC practice). High expression is reported in lymph-node germinal center cells and bone-marrow hematopoietic cells, whereas adipocytes are reported as not detected (HPA tissue IHC). A strong membrane or extracellular pattern conflicts with MCM5's nuclear localisation and lack of a transmembrane segment (UniProt P33992 localisation and topology). Check edge-restricted staining, necrotic areas, and endogenous enzyme signal with morphology and no-primary controls before calling cells positive (standard IHC practice). Finally, interpret heterogeneous nuclear staining in light of MCM5's role in DNA replication rather than assuming every cell should stain equally (UniProt P33992 function).
Boster reagents
Best MCM5 / DNA replication licensing factor MCM5 IHC Antibodies
Anti-MCM5 antibodies have IHC images from human paraffin sections and IF/ICC images from A431 and U20S cells (catalog image captions). Both listed products have Human, Mouse and Rat reactivity (catalog).
Real IHC data
Anti-MCM5 ® Antibody (monoclonal, 4G10)
Cat # M03642-2
Real IHC data
Anti-MCM5 Antibody ®
Cat # PA1653
M03642-2 has IHC images from human gallbladder adenocarcinoma, lung cancer and pancreatic cancer paraffin sections, plus an IF/ICC image from A431 cells (M03642-2 image captions). PA1653 has an IHC image from human intestinal cancer tissue and an IF/ICC image from U20S cells (PA1653 image captions).
Which to pick: For tissue IHC, M03642-2 offers a mouse monoclonal antibody with documented EDTA pH 8.0 retrieval and 2 μg/ml staining on human paraffin sections; the fixative is unreported (catalog: clone 4G10; M03642-2 IHC caption). For IF/ICC, PA1653 is a rabbit antibody with a documented 2 μg/ml U20S cell image; its separate intestinal cancer IHC caption does not report the fixative (catalog: PA1653 applications; PA1653 IF/ICC and IHC captions). For cross-species work, both list Human, Mouse and Rat reactivity, but the supplied IHC images for these two SKUs show human samples (catalog: reactivity; M03642-2 and PA1653 IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P33992 (MCM5_HUMAN, DNA replication licensing factor MCM5).
- Human Protein Atlas. MCM5 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. MCM5 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. MCM5 antibody validation summary (3 antibodies).
- Minichromosome maintenance protein 5 - a promising prognostic marker of oral epithelial dysplasias and oral squamous cell carcinomas. Journal of oral and maxillofacial pathology : JOMFP 2023 — PMC10829438.
- MCMs expression in lung cancer: implication of prognostic significance. Journal of Cancer 2017 — PMC5688916.
- Diagnosis of pancreaticobiliary malignancy by detection of minichromosome maintenance protein 5 in bile aspirates. British journal of cancer 2008 — PMC2391096.
- MCM5 Aggravates the HDAC1-Mediated Malignant Progression of Lung Cancer. Frontiers in cell and developmental biology 2021 — PMC8366603.
- PubMed PMID:15461802 — UniProt-cited evidence.
- PubMed PMID:10591208 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MCM5 (DNA replication licensing factor MCM5). Expected localisation Nuclear staining in selected tissue cells (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M03642-2). Validated IHC antibody: M03642-2 at 2μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Germinal center cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human gallbladder adenocarcinoma tissue; fixative not specified (datasheet M03642-2); 2. Fixation: Image fixative and duration unreported (datasheet M03642-2); 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 M03642-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M03642-2); 8. Primary antibody: Mouse monoclonal (clone 4G10) anti-MCM5, 2μg/ml (datasheet M03642-2); 9. Primary incubation: Overnight at 4 °C (datasheet M03642-2). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Chromatin association changes during DNA replication (UniProt); Isoform / epitope: No annotated isoforms; the chain spans residues 2–734 (UniProt); Regulation: Expression regulation is not annotated (UniProt). Evidence: UniProt P33992, HPA tissue IHC · Enhanced, HPA ICC-IF · nucleoplasm., Datasheet M03642-2, PMC10829438, PMC2391096. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MCM5/immunohistochemistry-ihc-guide.