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- Table of Contents
MARCO / Macrophage receptor MARCO · Western blot design guide
Design a Western Blot for MARCO
Real validated MARCO Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MARCO WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled September 2026 · For research use; verify linked source records and product datasheet before use
MARCO Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~52.7 kDa | |
| Observed band | Approximately 80 kDa | |
| Gel | 10% (catalog A00391-3) | |
| Positive control | Lung (IHC candidate; verify WB) +1 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Disulfide-linked | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated MARCO Western Blot Protocols
The A00391-3 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human THP-1, human THP-1(+LPS), human U2OS (catalog A00391-3) |
| Gel % | 10% (catalog A00391-3) |
| Load | 30 ug; reducing conditions (catalog A00391-3) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A00391-3) |
| Membrane | nitrocellulose membrane (catalog A00391-3) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A00391-3) |
| Primary antibody | A00391-3 · 0.5 μg/mL (catalog A00391-3) |
| Primary incubation | overnight at 4°C (catalog A00391-3) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A00391-3) |
| Secondary incubation | 1.5 hour at RT (catalog A00391-3) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A00391-3) |
| Detection | ECL (catalog A00391-3) |
Section 2
What Is the Expected MARCO Western Blot Band Size?
MARCO is predicted at 52.7 kDa and observed near 80 kDa under reducing conditions; the cause of that difference is not established.
| Band near 80 kDa | Empirical MARCO band observed under reducing conditions; the reason it exceeds the predicted mass is not established |
| Band near 53 kDa | Near the predicted 52.7 kDa sequence mass; confirm its identity with controls |
| High-mass band under non-reducing conditions | May represent disulfide-linked MARCO homotrimers or larger oligomers |
| Several bands at different positions | Isoforms 1 and 2 or oligomeric states may contribute; distinct isoform migration is unverified |
| Weak band in whole-cell lysate | MARCO is a cell-membrane receptor |
💡Expected MARCO appearanceMARCO has a predicted sequence mass of 52.7 kDa, while antibody QC detects approximately 80 kDa under reducing conditions; use a positive control and band-identity controls because the difference is unexplained.
| Predicted 52.7 kDa sequence mass | Provides the monomer reference; an empirical band appears near 80 kDa under reducing conditions |
| N-linked glycosylation at Asn83 | May affect apparent migration; its contribution to the 80 kDa band is unmeasured |
| N-linked glycosylation at Asn136 | May affect apparent migration; its contribution to the 80 kDa band is unmeasured |
| Disulfide-linked homotrimer | May retain a higher-mass species under non-reducing or incomplete reduction conditions |
| Isoforms 1 and 2 | May differ in size, but their individual masses and band separation are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated MARCO may be scarce in the sampled lysate | Check MARCO expression with a positive control and test a membrane-enriched fraction |
| Band higher than expected | The approximately 80 kDa empirical band exceeds the 52.7 kDa prediction; its cause is unestablished | Compare with a positive control and assess glycosylation or reduction effects separately |
| Broad smear instead of sharp band | Variable N-linked glycosylation is possible at Asn83 and Asn136 | Compare matched samples before and after N-glycan removal |
| Multiple bands | Isoforms 1 and 2 or incompletely reduced oligomers may contribute | Compare reducing and non-reducing lanes and verify bands with MARCO-specific controls |
| Weak or no signal | MARCO abundance in the sampled cells or membrane fraction may be low | Use an expression-confirmed positive control and enrich the membrane fraction |
Experiment evidence
Western blot images showing MARCO band
A00391-3 · MARCO
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MARCO Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for MARCO in Western blot, you can use lung tissue, which has high HPA expression.
Positive control: Lung (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: As a membrane protein, MARCO may be easier to detect in a membrane-enriched lysate.
HPA tissue expression evidence for MARCO
Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Higher expression tissues · candidate positive controls from IHC
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Lung | macrophages | High | Protein (IHC) | HPA → |
| Testis | spermatogonia cells | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-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 → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MARCO Western Blot Tips
Deeper troubleshooting and optimisation questions for MARCO, answered from its protein features.
How should MARCO band migration be interpreted?
Band shift · Use the separately labelled calculated mass and catalog-observed evidence above. A sequence annotation does not establish an observed migration shift. Verify target identity with orthogonal controls.
How could MARCO isoforms affect band interpretation?
Isoforms · UniProt lists isoforms 1 and 2. Isoform 2 lacks residues 1–78 in the canonical UniProt sequence, so it may yield a different band or escape detection by an antibody targeting that region. Confirm the antibody epitope before assigning bands to isoforms.
Choose an epitope outside canonical UniProt residues 1–78, which are missing from isoform 2. Check the antibody’s stated epitope before interpreting absence of a band as absence of MARCO.
Choose an epitope outside canonical UniProt residues 1–78, which are missing from isoform 2. Check the antibody’s stated epitope before interpreting absence of a band as absence of MARCO.
Which MARCO glycosylation sites matter for blot analysis?
PTM · UniProt lists N-linked glycosylation at asparagines 83 and 136, using canonical sequence coordinates. Compare matched samples before and after N-glycan removal to assess its effect on migration; the site annotations alone do not predict a visible shift.
Does this guide establish induction of MARCO?
Induction · No general induction response is established by this guide. A pathway or gene-set association is not evidence of induction in a particular specimen. Verify the relevant treatment and control in a target-specific experiment.
How should transfer be checked for MARCO?
Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A00391-3 datasheet where specified. Otherwise compare 5% milk or 5% BSA in TBST; for a phospho-specific assay start with BSA. Optimize background and specific signal with matched controls.
How should I quantify MARCO bands across samples?
Quantitation · Use the same sample preparation and reducing conditions across samples, then quantify corresponding bands consistently. MARCO’s two isoforms, N-linked glycosylation and disulfide-linked oligomers can complicate assignment of different bands to one protein form.
Why might MARCO appear near 80 kDa instead of 52.7 kDa?
Interpretation · The supplied observed band is approximately 80 kDa, while the predicted mass is 52.7 kDa. MARCO has N-linked glycosylation sites at UniProt positions 83 and 136, but their presence alone does not establish the cause or size of the apparent shift. Compare untreated and deglycosylated samples to test whether glycans contribute.
UniProt describes MARCO as a disulfide-linked homotrimer that may form larger oligomers and lists three disulfide bonds. Compare reducing and nonreducing samples when assessing high-molecular-weight bands. A band’s position alone cannot establish its oligomeric state.
Check whether the antibody recognizes canonical residues 1–78, compare reducing with nonreducing samples, and assess the effect of N-glycan removal. These checks address the listed isoform difference, disulfide-linked oligomers and glycosylation sites without assigning an unexpected band from size alone.
UniProt describes MARCO as a disulfide-linked homotrimer that may form larger oligomers and lists three disulfide bonds. Compare reducing and nonreducing samples when assessing high-molecular-weight bands. A band’s position alone cannot establish its oligomeric state.
Check whether the antibody recognizes canonical residues 1–78, compare reducing with nonreducing samples, and assess the effect of N-glycan removal. These checks address the listed isoform difference, disulfide-linked oligomers and glycosylation sites without assigning an unexpected band from size alone.
Boster reagents
MARCO Western Blot Antibodies
Catalog antibodies with Western blot application and product-specific WB images. Evaluate suitability with the reported sample, controls and experimental conditions.
Real WB data
Anti-MARCO Antibody Picoband®
Cat # A00391-3
The catalog reports one anti-MARCO antibody for Western blot: A00391-3. Its image shows human cell, rat spleen, and mouse cell lysates tested at 0.5 μg/mL. The reported band is approximately 80 kDa, versus an expected 53 kDa; no independent validation is supplied.
Which to pick: A00391-3 is the only listed option. It has a Western blot image covering human THP-1 and U2OS cells, rat spleen, and mouse RAW264.7 cells. Consider the 80 kDa observed band versus the 53 kDa expected size when interpreting your result.
Source: BosterBio MARCO gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9UEW3.
- Human Protein Atlas. MARCO tissue expression.
- PMC9200231 — target-verified WB comparison
- PMC12936005 — target-verified WB comparison
- PMC9667514 — target-verified WB comparison