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- Table of Contents
TMEM59L / Transmembrane protein 59-like · Western blot design guide
Design a Western Blot for TMEM59L
Source-linked TMEM59L Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TMEM59L 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
TMEM59L 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 | ~37.6 kDa | |
| Observed band | 68 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Caudate (IHC candidate; verify WB) +3 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Cleaved | |
| Caveat | Band identity controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked TMEM59L Western Blot Protocol Options
The A15285 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | rat heart tissue lysate (catalog A15285) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A15285 · 1 μg/mL (catalog A15285) |
| 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 TMEM59L Western Blot Band Size?
TMEM59L is predicted at 37.6 kDa, while antibody QC reports 68 kDa; the cause of the difference is not established.
| Band near 68 kDa | Empirical TMEM59L band reported by antibody QC; confirm identity with controls |
| Band near 37.6 kDa | Compatible with the calculated full-length precursor mass; identity requires confirmation |
| Band below the precursor position | Could reflect removal of the residues 1–24 signal peptide |
| Band above the calculated precursor position | N-linked glycosylation at Asn97 could affect migration, but the size effect is unmeasured |
| Little signal in a soluble fraction | TMEM59L is a Golgi membrane protein and may partition with membranes |
💡Expected TMEM59L appearanceTMEM59L has a predicted full-length mass of 37.6 kDa, while antibody QC reports a 68 kDa band; the difference is unexplained, so confirm band identity with appropriate controls.
| Predicted full-length mass | 37.6 kDa is the calculated precursor mass; antibody QC reports a 68 kDa band whose difference is unexplained |
| N-linked glycosylation at Asn97 | May affect apparent migration, but the size and visibility of any shift are unestablished |
| Signal peptide at residues 1–24 | Contributes to the calculated precursor mass before cleavage |
| Signal peptide cleavage | Would make the mature chain smaller than the full-length precursor; its apparent mass is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Golgi membrane localization may limit recovery in the prepared fraction | Check membrane extraction and test a membrane-enriched fraction |
| Band higher than expected | The reported 68 kDa band exceeds the 37.6 kDa prediction, with no established cause | Confirm identity using an independent antibody or TMEM59L depletion; compare migration after deglycosylation |
| Band lower than expected | Signal peptide cleavage could lower mass relative to the full-length precursor | Check antibody epitope coverage and confirm identity with TMEM59L depletion |
| Broad smear instead of sharp band | Variable modification at the documented N-linked site is possible but unproven | Compare untreated and deglycosylated samples, then check whether the signal tracks with TMEM59L depletion |
| Multiple bands | Signal peptide processing or modification at Asn97 may contribute, but distinct bands are unverified | Compare deglycosylated samples and use TMEM59L depletion to identify specific bands |
| Weak or no signal | Membrane protein recovery or detection may be insufficient | Check extraction, loading, and a validated positive lysate |
Experiment evidence
Western blot images showing TMEM59L band
A15285 · TMEM59L
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TMEM59L Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TMEM59L in Western blot, you can use caudate tissue, which HPA reports as having medium expression.
Positive control: Caudate (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside TMEM59L.
⚠️Feasibility: As a Golgi membrane protein, TMEM59L may require lysis that retains membrane proteins for a clear signal.
HPA tissue expression evidence for TMEM59L
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 |
|---|---|---|---|---|
| Caudate | neuronal cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal cells | Medium | Protein (IHC) | HPA → |
| Hippocampus | neuronal 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 TMEM59L Western Blot Tips
Deeper troubleshooting and optimisation questions for TMEM59L, answered from its protein features.
How should TMEM59L 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 multiple bands represent annotated TMEM59L isoforms?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore does not support assigning additional bands to annotated TMEM59L isoforms. Evaluate any extra bands independently before including them in analysis.
Which glycosylation site matters when interpreting TMEM59L bands?
PTM · UniProt lists one N-linked glycosylation site, Asn97, in UniProt sequence numbering. This makes glycosylation relevant when investigating band mobility, but the annotation does not establish a visible shift or account for the 68 kDa band by itself.
Does this guide establish induction of TMEM59L?
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 TMEM59L?
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 A15285 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 TMEM59L be quantified across Western blots?
Quantitation · Measure the same validated TMEM59L band across samples and state which apparent mass was measured. Keep the 68 kDa observed position distinct from the 37.6 kDa predicted mass; the listed signal peptide and Asn97 glycosylation site do not establish which band is suitable for quantitation.
Does the 68 kDa band match TMEM59L's predicted mass?
Interpretation · The supplied apparent band is 68 kDa, versus a predicted mass of 37.6 kDa. TMEM59L has a signal peptide at residues 1–24 and an N-linked glycosylation site at Asn97, using UniProt numbering. These annotations do not establish the cause or size of the difference. Confirm band identity before treating 68 kDa as TMEM59L.
UniProt annotates a signal peptide at residues 1–24. Its removal would shorten the protein, so this feature alone does not explain a band above the 37.6 kDa predicted mass. Do not assign a processing state from band position alone.
TMEM59L is annotated as a Golgi apparatus membrane protein with single-pass type I topology. When comparing fractions, account for their Golgi membrane content. Localization can guide interpretation of where TMEM59L is expected, but it does not identify an unexpected band or explain its apparent mass.
UniProt annotates a signal peptide at residues 1–24. Its removal would shorten the protein, so this feature alone does not explain a band above the 37.6 kDa predicted mass. Do not assign a processing state from band position alone.
TMEM59L is annotated as a Golgi apparatus membrane protein with single-pass type I topology. When comparing fractions, account for their Golgi membrane content. Localization can guide interpretation of where TMEM59L is expected, but it does not identify an unexpected band or explain its apparent mass.
Boster reagents
TMEM59L 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 59-like TMEM59L Antibody
Cat # A15285
The catalog reports one anti-TMEM59L antibody, A15285, with reported Human, Mouse, and Rat reactivity. Its Western blot image shows rat heart tissue lysate tested at 1 μg/mL; the supplied evidence does not show WB results for human or mouse samples.
Which to pick: A15285 is the only listed option. Its WB image documents testing in rat heart lysate at 1 μg/mL. For human or mouse samples, reactivity is listed, but no WB example is supplied.
Source: BosterBio TMEM59L gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9UK28.
- Human Protein Atlas. TMEM59L tissue expression.
- PMC13579383 — target-verified WB comparison
- PMC12809716 — target-verified WB comparison
- PMC5288309 — target-verified WB comparison