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- Table of Contents
MIA / Melanoma-derived growth regulatory protein · Western blot design guide
Design a Western Blot for MIA
Real validated MIA Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MIA 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
MIA 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 | ~14.5 kDa | |
| Gel | 15% (standard starting point) | |
| Positive control | Hippocampus (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 | Cleaved | |
| Caveat | Secreted protein | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated MIA Western Blot Protocols
The A01570 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | BV2 (40ug), C6 (40ug), K562 (40ug) (catalog A01570) |
| Gel % | 15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.2 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A01570; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected MIA Western Blot Band Size?
UniProt predicts a 14.5 kDa precursor; secretion, signal-peptide cleavage, and isoforms may affect the blot, but their migration effects are unconfirmed.
| Band near 14.5 kDa | may represent the full-length precursor; confirm its identity |
| Band below 14.5 kDa | may represent protein after signal-peptide cleavage |
| Little or no band in whole-cell lysate | consistent with secretion |
| Several discrete bands | could include isoforms 1 and 2 or precursor and mature protein; identities require validation |
💡Expected MIA appearanceUniProt predicts a 14.5 kDa precursor; signal-peptide cleavage may yield a smaller mature band, but no empirical band size or isoform migration is established, so confirm band identity with controls.
| UniProt predicted precursor mass | 14.5 kDa before signal-peptide cleavage |
| Signal peptide at residues 1–24 | cleavage can make the mature protein smaller than the precursor |
| Isoform 1 | its mass relative to isoform 2 is not supplied |
| Isoform 2 | its mass relative to isoform 1 is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | MIA is secreted | check conditioned medium alongside a positive control |
| Band higher than expected | an uncleaved precursor may run above mature MIA | compare lysate with conditioned medium and verify band identity |
| Band lower than expected | signal-peptide cleavage may lower the mass from the 14.5 kDa precursor | check conditioned medium and confirm identity with a positive control |
| Multiple bands | isoforms 1 and 2 or precursor and mature protein may contribute | compare lysate and conditioned medium and use an identity control |
| Weak or no signal | secretion may leave little MIA in the sampled lysate | test conditioned medium and a positive control |
Experiment evidence
Western blot images showing MIA band
A01570 · MIA
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MIA Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for MIA in Western blot, you can use hippocampus tissue, which HPA lists as a positive sample.
Positive control: Hippocampus (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: Because MIA is secreted, signal in tissue lysate may be weak; consider an extracellular protein fraction.
HPA tissue expression evidence for MIA
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 |
|---|---|---|---|---|
| Hippocampus | synapses | Medium | Protein (IHC) | HPA → |
| Testis | round or early spermatids | 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 MIA Western Blot Tips
Deeper troubleshooting and optimisation questions for MIA, answered from its protein features.
How should MIA 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 MIA isoforms affect the band pattern?
Isoforms · UniProt lists two isoforms. In isoform 2, residues 43..131 of isoform 1 are replaced by RSGRLLWRSGCSPGLFPQ. Check which sequence your antibody recognizes before assigning bands to either isoform; the features alone do not establish where either band runs.
Determine whether the antibody recognizes both isoforms or only one. The isoforms differ across UniProt residues 43..131, so an epitope in that region could make band intensity reflect antibody recognition as well as protein abundance.
Determine whether the antibody recognizes both isoforms or only one. The isoforms differ across UniProt residues 43..131, so an epitope in that region could make band intensity reflect antibody recognition as well as protein abundance.
Do annotated modifications prove a band shift?
PTM · The linked UniProt record describes protein features. A modification annotation alone does not demonstrate a visible shift; retain any condition or experimental qualifier attached to it.
Does this guide establish induction of MIA?
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 MIA?
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 A01570 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 MIA be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
Why might MIA migrate differently from its predicted 14.5 kDa mass?
Interpretation · MIA has a signal peptide at UniProt residues 1..24 and is secreted, so processing could change the mass of the detected protein. The supplied features do not establish a visible band shift or a specific apparent mass.
MIA is annotated as secreted. Check conditioned medium when a cellular lysate gives a weak signal, and compare equivalent collection conditions across samples.
UniProt lists two disulfide bonds. Comparing reducing and nonreducing lanes may help assess whether disulfide-dependent structure affects migration. These features do not guarantee a detectable change.
Consider signal-peptide processing, the two isoforms, and the two disulfide bonds when evaluating band size. MIA also interacts with FASLG and TMIGD2, but those interactions alone do not establish that an unexpected band is a complex. Confirm band identity before assigning a cause.
MIA is annotated as secreted. Check conditioned medium when a cellular lysate gives a weak signal, and compare equivalent collection conditions across samples.
UniProt lists two disulfide bonds. Comparing reducing and nonreducing lanes may help assess whether disulfide-dependent structure affects migration. These features do not guarantee a detectable change.
Consider signal-peptide processing, the two isoforms, and the two disulfide bonds when evaluating band size. MIA also interacts with FASLG and TMIGD2, but those interactions alone do not establish that an unexpected band is a complex. Confirm band identity before assigning a cause.
Boster reagents
MIA 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-MIA Antibody
Cat # A01570
A01570 is catalogued for human, mouse, and rat MIA. Its Western blot image shows testing at 1:500 on BV2, C6, K562, A2780, U-87MG, and HEK293T whole-cell lysates (40 µg each). Broader validation is not supplied.
Which to pick: A01570 is the only listed anti-MIA antibody. Its WB image documents six cell lysates at 1:500; compare those tested samples and the stated reactivity with your experiment.
Source: BosterBio MIA gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q16674.
- Human Protein Atlas. MIA tissue expression.
- PMC551595 — target-verified WB comparison
- PMC9856983 — target-verified WB comparison