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- Table of Contents
TMEM106A / Transmembrane protein 106A · Western blot design guide
Design a Western Blot for TMEM106A
Source-linked TMEM106A Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TMEM106A WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled October 2026 · For research use; verify linked source records and product datasheet before use
TMEM106A 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 | ~28.9 kDa | |
| Observed band | 68 kDa | |
| Gel | 12–15% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | Blocking peptide control | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked TMEM106A Western Blot Protocol Options
The A16224 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | A-20 cell lysate (catalog A16224) |
| Gel % | 12–15% (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 | 0.45 µm PVDF (standard starting point) |
| Blocking | Western blot analysis of TMEM106A in A-20 cell lysate with TMEM106A antibody at 1 μg/mL in (A) the absence and (B) the presence of blocking peptide (catalog A16224) |
| Primary antibody | A16224 · 1 μg/mL (catalog A16224) |
| 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 TMEM106A Western Blot Band Size?
TMEM106A is predicted at 28.9 kDa; antibody QC reports 68 kDa, but the cause of the difference is not established.
| Band near 68 kDa | Empirical antibody QC band; TMEM106A identity requires controls |
| Band near 28.9 kDa | Near the UniProt predicted polypeptide mass |
| Several bands at different positions | Isoforms 1 and 2 are documented, but distinct migration is unverified |
| Little or no band in a soluble fraction | TMEM106A is a cell membrane protein |
💡Expected TMEM106A appearanceUniProt predicts 28.9 kDa, while antibody QC reports a 68 kDa band; the cause of this difference is unestablished, so confirm band identity with appropriate controls.
| UniProt predicted mass | Predicts 28.9 kDa; the reported 68 kDa band has an unexplained difference |
| Isoform 1 | Its individual mass and migration are not supplied |
| Isoform 2 | Its individual mass and migration are not supplied |
| Alternative splicing | Produces isoforms 1 and 2, whose size difference is unreported |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane protein may be poorly recovered during extraction | Check the membrane fraction and extraction conditions |
| Band higher than expected | The reported 68 kDa band exceeds the 28.9 kDa prediction for an unknown reason | Confirm identity with blocking peptide and an independent antibody |
| Band lower than expected | Identity or isoform migration is uncertain | Check antibody specificity and compare with a reference sample |
| Multiple bands | Two isoforms exist, but distinct bands have not been demonstrated | Check each band with an independent antibody or isoform-specific evidence |
| Weak or no signal | Membrane extraction or antibody detection may be inadequate | Check membrane recovery, loading, and antibody performance |
Experiment evidence
Western blot images showing TMEM106A band
A16224 · TMEM106A
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TMEM106A Western blot
🧪For positive controls for TMEM106A in Western blot, you can use an HPA-IHC candidate positive sample once one is identified; none is supplied here.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the blot.
⚠️Feasibility: HPA data are unavailable, so positive and negative tissue controls cannot be selected from the supplied evidence.
HPA tissue expression evidence for TMEM106A
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced TMEM106A Western Blot Tips
Deeper troubleshooting and optimisation questions for TMEM106A, answered from its protein features.
How should TMEM106A 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 TMEM106A isoforms produce different bands?
Isoforms · Two isoforms are listed. In isoform 2, canonical residues 1–49 are replaced by a single methionine, so its sequence is shorter. These are UniProt canonical coordinates; antibody or paper numbering may differ. The sequence change alone does not establish where either isoform will migrate.
Check its epitope against both sequences. An antibody targeting canonical residues 1–49 may miss isoform 2 because that segment is replaced by one methionine. An epitope retained in both isoforms is better suited to measuring both.
Check its epitope against both sequences. An antibody targeting canonical residues 1–49 may miss isoform 2 because that segment is replaced by one methionine. An epitope retained in both isoforms is better suited to measuring both.
Should the 68 kDa band be attributed to a modification?
PTM · The supplied record lists no modified residues or glycosylation sites. It therefore provides no specific modification to assign to the 68 kDa band. Feature annotations alone cannot explain the difference from the predicted mass.
Does this guide establish induction of TMEM106A?
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 TMEM106A Western blot?
Transfer · TMEM106A is annotated as a single-pass cell-membrane protein. The supplied features do not specify a transfer method. Check transfer efficiency for the chosen conditions before interpreting a weak or absent band.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A16224 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 TMEM106A bands be quantified?
Quantitation · Define which validated band and isoform the measurement represents. Because TMEM106A is annotated at the cell membrane, use consistent sample preparation across lanes and normalize within the same preparation. Report the band measured rather than combining unresolved bands.
Why might TMEM106A appear at 68 kDa rather than 28.9 kDa?
Interpretation · The supplied Western blot reports 68 kDa, while the predicted mass is 28.9 kDa. The listed features do not establish the cause of that difference. Confirm the band with an independent antibody or TMEM106A depletion before assigning it to TMEM106A.
Consider the two listed isoforms and whether the antibody epitope is retained in isoform 2. Compare candidate bands with the reported 68 kDa band, then use an independent antibody or TMEM106A depletion to test their identity. Do not identify a band from predicted mass alone.
Consider the two listed isoforms and whether the antibody epitope is retained in isoform 2. Compare candidate bands with the reported 68 kDa band, then use an independent antibody or TMEM106A depletion to test their identity. Do not identify a band from predicted mass alone.
Boster reagents
TMEM106A 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-Transmembrane protein 106A TMEM106A Antibody
Cat # A16224
The catalog reports one anti-TMEM106A antibody for Western blot, A16224, with reported human and mouse reactivity. Its WB image shows A-20 cell lysate tested at 1 μg/mL with and without blocking peptide. The supplied evidence does not establish performance across other samples.
Which to pick: A16224 is the only listed option. Its WB image documents an A-20 cell lysate experiment at 1 μg/mL with a blocking peptide comparison; check whether those conditions fit your sample.
Source: BosterBio TMEM106A gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q96A25.
- Human Protein Atlas. TMEM106A tissue expression.
- PMC9827947 — target-verified WB comparison
- PMC4190911 — target-verified WB comparison
- PMC8963425 — target-verified WB comparison