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- Table of Contents
IMMT / MICOS complex subunit MIC60 · Western blot design guide
Design a Western Blot for IMMT
Source-linked IMMT Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-IMMT 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
IMMT Western Blot Experimental Design Guide
Expected bands, source-linked protocol options, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~83.7 kDa | |
| Observed band | ~80–90 kDa | |
| Gel | 5–20% (catalog A04102-2) | |
| Positive control | Heart muscle (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 4 isoform(s) |
Section 1
Source-Linked IMMT Western Blot Protocol Options
The A04102-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Colo320, human HepG2, rat NRK, rat PC-12, rat C6, mouse HEPA1/6, mouse MFC (catalog A04102-2) |
| Gel % | 5–20% (catalog A04102-2) |
| Load | 30 ug; reducing conditions (catalog A04102-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A04102-2) |
| Membrane | nitrocellulose membrane (catalog A04102-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A04102-2) |
| Primary antibody | A04102-2 · 0.5 μg/mL (catalog A04102-2) |
| Primary incubation | overnight at 4°C (catalog A04102-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A04102-2) |
| Secondary incubation | 1.5 hour at RT (catalog A04102-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A04102-2) |
| Detection | ECL (catalog A04102-2) |
Section 2
What Is the Expected IMMT Western Blot Band Size?
IMMT is predicted at 83.7 kDa and observed at ~80–90 kDa; the cause of any difference is not established.
| Band at ~80–90 kDa | Empirical IMMT signal near the predicted 83.7 kDa; confirm identity with controls |
| Single band near 83.7 kDa | Consistent with predicted IMMT mass |
| Additional discrete bands | Could represent isoforms 1–4, whose migration is unestablished |
| Weak or absent band in a mitochondria-depleted fraction | Consistent with mitochondrial inner-membrane localization |
💡Expected IMMT appearanceIMMT has a predicted mass of 83.7 kDa and an empirical band at ~80–90 kDa; confirm band identity with ordinary controls because isoform-specific migration is unknown.
| Predicted IMMT mass | 83.7 kDa predicted; an empirical band appears at ~80–90 kDa |
| Splice isoform 1 | Its migration relative to the other isoforms is unknown |
| Splice isoform 2 | Its migration relative to the other isoforms is unknown |
| Splice isoform 3 | Its migration relative to the other isoforms is unknown |
| Splice isoform 4 | Its migration relative to the other isoforms is unknown |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Mitochondrial inner-membrane IMMT may be poorly recovered | Check mitochondrial recovery and membrane-protein extraction |
| Band higher than expected | The band's identity or migration is unestablished | Compare with the ~80–90 kDa reference and verify identity by IMMT depletion |
| Band lower than expected | An isoform is possible, but its migration is unknown | Verify identity by IMMT depletion before assigning an isoform |
| Multiple bands | Four isoforms are listed, but distinct bands are unproven | Check which bands decrease after IMMT depletion |
| Weak or no signal | Mitochondrial inner-membrane protein may be poorly extracted | Check extraction and a mitochondrial loading marker |
Experiment evidence
Western blot images showing IMMT band
A04102-2 · IMMT
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for IMMT Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for IMMT in Western blot, you can use heart muscle lysate, based on high HPA expression.
Positive control: Heart muscle (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: Heart muscle offers a strong positive sample, while HPA lists adipose tissue as not detected.
HPA tissue expression evidence for IMMT
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 |
|---|---|---|---|---|
| Heart muscle | cardiomyocytes | High | Protein (IHC) | HPA → |
| Kidney | collecting ducts | High | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | High | Protein (IHC) | HPA → |
| Stomach | glandular cells | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular 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 → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Ovary | follicle cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced IMMT Western Blot Tips
Deeper troubleshooting and optimisation questions for IMMT, answered from its protein features.
How should IMMT 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.
Could IMMT isoforms produce different bands?
Isoforms · UniProt lists four isoforms. Relative to the canonical sequence, isoform 2 lacks residues 142–152, isoform 3 replaces residues 187–219 with A, and isoform 4 lacks residue 188. These differences could affect migration or antibody recognition, depending on the epitope; the features alone do not predict distinct visible bands.
Which IMMT phosphorylation sites matter when interpreting a band?
PTM · UniProt lists phosphoserines at canonical positions 103, 112, 113, 223, 388, and 390. Check whether an antibody’s epitope overlaps these sites when comparing signals. Site presence alone does not establish a detectable mobility shift; paper or antibody numbering may use a different sequence convention.
N6-acetyllysine is listed at canonical positions 211, 222, and 451. Position 211 lies within the region replaced in isoform 3. When comparing site-specific results, check which isoform and numbering convention the antibody or paper uses. These annotations do not establish a visible band shift.
N6-acetyllysine is listed at canonical positions 211, 222, and 451. Position 211 lies within the region replaced in isoform 3. When comparing site-specific results, check which isoform and numbering convention the antibody or paper uses. These annotations do not establish a visible band shift.
Does this guide establish induction of IMMT?
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.
What transfer method to use for IMMT Western blot?
Transfer · IMMT is a single-pass mitochondrial inner-membrane protein with a predicted mass of 83.7 kDa. Choose transfer conditions that retain and transfer proteins in this size range, then check the membrane and post-transfer gel for recovery. The supplied features do not specify a particular transfer method or duration.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A04102-2 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 IMMT bands be quantified?
Quantitation · Quantify a consistently defined band in the reported 80–90 kDa range, keeping the same band boundaries across samples. Because four isoforms are listed, state whether the antibody and analysis measure a shared signal or a particular isoform. The features do not establish that every band in the range represents the same species.
Why might IMMT migrate around 80–90 kDa?
Interpretation · The reported band range is close to IMMT’s predicted 83.7 kDa. The listed modifications and isoforms do not, by themselves, establish a visible shift or explain a difference between apparent and calculated mass.
First compare their positions with the 80–90 kDa reported range and the predicted 83.7 kDa mass. Consider the listed isoform sequence changes and whether the antibody epitope is retained. IMMT’s phosphorylation and acetylation annotations alone cannot identify an unexpected band or prove its cause.
First compare their positions with the 80–90 kDa reported range and the predicted 83.7 kDa mass. Consider the listed isoform sequence changes and whether the antibody epitope is retained. IMMT’s phosphorylation and acetylation annotations alone cannot identify an unexpected band or prove its cause.
Boster reagents
IMMT 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-Mitofilin/IMMT Antibody Picoband®
Cat # A04102-2
The catalog reports one anti-IMMT antibody, A04102-2, with reported human, mouse, and rat reactivity. Its WB image shows an approximately 80–90 kDa band in the specified whole-cell lysates. The supplied evidence does not establish performance in other sample types.
Which to pick: A04102-2 is the only listed option and has a WB image using human Colo320 and HepG2, rat NRK, PC-12 and C6, and mouse HEPA1/6 and MFC lysates. Use those tested contexts to guide selection; optimize for your sample.
Source: BosterBio IMMT gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q16891.
- Human Protein Atlas. IMMT tissue expression.
- PMC11150385 — target-verified WB comparison
- PMC13081484 — target-verified WB comparison
- PMC5928897 — target-verified WB comparison