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- Table of Contents
TMEM135 / Transmembrane protein 135 · Western blot design guide
Design a Western Blot for TMEM135
Source-linked TMEM135 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TMEM135 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
TMEM135 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 | ~52.3 kDa | |
| Observed band | 68 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Adipose tissue (IHC candidate; verify WB) +4 more | |
| Negative control | Bone marrow (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | Observed above predicted | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked TMEM135 Western Blot Protocol Options
The A14036 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | rat liver tissue lysate (catalog A14036) |
| 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 | A14036 · (A) 1 and (B) 2 μg/mL (catalog A14036) |
| 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 TMEM135 Western Blot Band Size?
TMEM135 is predicted at 52.3 kDa and observed at 68 kDa; the cause of this difference is not established.
| Band at 68 kDa | Empirical TMEM135 band; its difference from the 52.3 kDa prediction has no established cause here |
| Band near 52.3 kDa | Near the predicted protein mass; confirm identity with controls |
| Bands at different positions | Isoforms 1 and 2 are possible contributors, but their migration is unknown |
| Weak or absent band in a soluble fraction | TMEM135 is a multi-pass mitochondrial and peroxisomal membrane protein |
💡Expected TMEM135 appearanceUniProt predicts 52.3 kDa, while antibody QC reports a 68 kDa band; the cause of the difference is unknown, so confirm band identity with appropriate controls.
| UniProt predicted mass of 52.3 kDa | The empirical 68 kDa band migrates above this prediction for an unknown reason |
| Isoform 1 | Its individual mass and migration are not supplied |
| Isoform 2 | Its individual mass and migration are not supplied |
| Alternative splicing of isoforms 1 and 2 | Could affect band position, but a resolvable difference is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Multi-pass membrane localization may limit recovery | Check membrane extraction and a mitochondrial or peroxisomal fraction |
| Band higher than expected | The observed 68 kDa band exceeds the 52.3 kDa prediction for an unknown reason | Confirm identity using knockdown or an independent antibody |
| Band lower than expected | An isoform may differ in size, but its migration is unknown | Check band identity and compare isoform-specific expression if available |
| Multiple bands | Isoforms 1 and 2 are possible contributors, without established band positions | Test band identity with knockdown or isoform-specific expression |
| Weak or no signal | TMEM135 may be poorly recovered from membrane-containing material | Assess membrane protein extraction and loading |
Experiment evidence
Western blot images showing TMEM135 band
A14036 · TMEM135
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TMEM135 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TMEM135 in Western blot, you can use adipose tissue, an HPA medium-expression sample.
Positive control: Adipose tissue (IHC candidate; verify WB)
Negative control: Bone marrow (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: As a multi-pass membrane protein, TMEM135 may be harder to detect in whole-tissue lysate.
HPA tissue expression evidence for TMEM135
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 |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Caudate | neuronal cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal cells | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | Not detected | Protein (IHC) | HPA → |
| Pancreas | exocrine glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TMEM135 Western Blot Tips
Deeper troubleshooting and optimisation questions for TMEM135, answered from its protein features.
How should TMEM135 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 the two isoforms produce different bands?
Isoforms · Isoform 2 lacks residues 133–154 relative to isoform 1, using the supplied UniProt coordinates. This sequence difference could affect mass, but it does not establish that either isoform produces a distinct visible band. Check which isoform the antibody recognizes.
Does an annotated modification explain the 68 kDa band?
PTM · No glycosylation sites or modified residues are listed for TMEM135. The supplied features therefore do not support assigning the 68 kDa band to a specific modification. They also do not establish what causes the difference from the predicted mass.
Does this guide establish induction of TMEM135?
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 TMEM135 Western blot?
Transfer · TMEM135 is annotated as a multi-pass membrane protein, but these features do not specify a transfer method. Compare transfer conditions using the same sample and assess recovery of the candidate band and protein left in the gel.
How should blocking be optimized for TMEM135?
Blocking · The supplied features do not identify a preferred blocking reagent. Compare blockers with the same antibody and membrane preparation, then choose the condition that gives a clear candidate band with low background.
How should TMEM135 bands be quantified across samples?
Quantitation · Use comparable sample preparations and measure the candidate band within the assay's linear range. TMEM135 is annotated at mitochondrial and peroxisomal membranes and has two isoforms, so keep fractionation and the band selected for measurement consistent across samples.
Why might TMEM135 run at 68 kDa instead of 52.3 kDa?
Interpretation · The reported apparent band is 68 kDa, while the predicted mass is 52.3 kDa. TMEM135 is a multi-pass membrane protein, but the supplied features do not establish why the band runs higher. Confirm band identity before assigning a cause to the difference.
Consider whether the antibody recognizes both isoforms: isoform 2 lacks UniProt residues 133–154. The supplied features do not show that this difference creates a visible second band, or that the 68 kDa band is TMEM135. Verify band identity before labeling additional bands as isoforms or modified protein.
Consider whether the antibody recognizes both isoforms: isoform 2 lacks UniProt residues 133–154. The supplied features do not show that this difference creates a visible second band, or that the 68 kDa band is TMEM135. Verify band identity before labeling additional bands as isoforms or modified protein.
Boster reagents
TMEM135 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 135 TMEM135 Antibody
Cat # A14036
The catalog reports A14036 for TMEM135 Western blotting, with stated human, mouse, and rat reactivity. Its WB image shows rat liver tissue lysate tested at 1 and 2 μg/mL; the supplied evidence does not show WB results in human or mouse samples.
Which to pick: A14036 is the only listed option. Its rat liver WB image provides a starting point for rat tissue experiments; human and mouse reactivity is listed, but no WB example for those species is supplied.
Source: BosterBio TMEM135 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q86UB9.
- Human Protein Atlas. TMEM135 tissue expression.
- PMC10541902 — target-verified WB comparison