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- Table of Contents
TM4SF1 / Transmembrane 4 L6 family member 1 · Western blot design guide
Design a Western Blot for TM4SF1
Source-linked TM4SF1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TM4SF1 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
TM4SF1 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 | ~21.6 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Adrenal gland (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 | Glycosylated | |
| Caveat | Glycosylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked TM4SF1 Western Blot Protocol Options
The A10368 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human lung tissue lysate (catalog A10368) |
| 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 | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A10368 · (A) 0.5 and (B) 1 μg/mL (catalog A10368) |
| 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 TM4SF1 Western Blot Band Size?
TM4SF1 is predicted at 21.6 kDa; N-linked glycosylation and tumor-cell complexes could affect migration, but no empirical band size or shift is established.
| Band near 21.6 kDa | consistent with the predicted TM4SF1 mass; identity requires confirmation |
| Band above 21.6 kDa | could reflect N-linked glycosylation at Asn129 or Asn159; a shift is not established |
| High molecular weight signal in tumor cells | could reflect the reported TM4SF1-containing complexes if they persist during sample preparation |
| Faint or absent lysate band | could reflect poor recovery of this multi-pass membrane protein |
💡Expected TM4SF1 appearanceTM4SF1 has a predicted mass of 21.6 kDa, but no empirical band size is supplied; confirm any candidate band with ordinary identity controls because its glycosylation and complexes do not establish migration.
| Predicted TM4SF1 mass | 21.6 kDa is the sequence-based reference, not a measured band |
| N-linked glycosylation at Asn129 | could increase apparent size if occupied; no shift is established |
| N-linked glycosylation at Asn159 | could increase apparent size if occupied; no shift is established |
| N-linked glycosylation at Asn129 and Asn159 | combined occupancy could affect migration, but its magnitude is unknown |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | poor extraction of multi-pass membrane TM4SF1 | check membrane-protein recovery and include a positive-control lysate |
| Band higher than expected | possible N-linked glycosylation or persistent tumor-cell complexes | compare deglycosylated and fully denatured samples; confirm identity |
| Band lower than expected | migration or glycosylation state cannot be assigned from the supplied features | confirm identity with an independent antibody or TM4SF1 depletion |
| Multiple bands | possible glycosylation states or retained complexes; distinct forms are not established | compare deglycosylated samples and confirm bands by TM4SF1 depletion |
| Weak or no signal | limited recovery of membrane-associated TM4SF1 | check extraction, loading, and a positive-control lysate |
Experiment evidence
Western blot images showing TM4SF1 band
A10368 · TM4SF1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TM4SF1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TM4SF1 in Western blot, you can use adrenal gland tissue.
Positive control: Adrenal gland (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: As a multi-pass membrane protein, TM4SF1 may require a membrane-enriched lysate for a clear signal.
HPA tissue expression evidence for TM4SF1
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | glandular cells | Medium | Protein (IHC) | HPA → |
| Colon | endothelial cells | Medium | Protein (IHC) | HPA → |
| Fallopian tube | glandular cells | Medium | Protein (IHC) | HPA → |
| Lung | alveolar 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 → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TM4SF1 Western Blot Tips
Deeper troubleshooting and optimisation questions for TM4SF1, answered from its protein features.
How should TM4SF1 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 TM4SF1 isoforms explain multiple bands?
Isoforms · The supplied features list one isoform and no alternative sequence. They do not support assigning multiple bands to distinct TM4SF1 isoforms.
Which TM4SF1 glycosylation sites should I consider?
PTM · UniProt annotates N-linked glycosylation at Asn129 and Asn159. These are UniProt sequence coordinates; paper or antibody numbering may differ. Compare treated and untreated samples if testing whether glycosylation affects a band.
Does this guide establish induction of TM4SF1?
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 TM4SF1 Western blot?
Transfer · TM4SF1 is a 202-residue multi-pass membrane protein. The supplied features do not establish a specific transfer method. Check whether TM4SF1 remains in the gel or passes through the membrane while optimizing transfer empirically.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A10368 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 I quantify TM4SF1 bands?
Quantitation · Use a consistent sample preparation and quantify comparable bands across samples. TM4SF1 is a membrane glycoprotein reported in high molecular weight complexes in tumor cells, so document which band or bands were included in the measurement.
Why might TM4SF1 migrate differently from its predicted mass?
Interpretation · TM4SF1 has a predicted mass of 21.6 kDa and two annotated N-linked glycosylation sites, at UniProt positions 129 and 159. These features alone do not establish a visible shift or explain a particular apparent mass; no observed band size was supplied.
UniProt reports TM4SF1 in high molecular weight complexes in tumor cells and notes interaction with SDCBP2. This makes a complex worth considering, but these features do not identify any particular Western-blot band.
Consider the annotated N-linked sites at UniProt positions 129 and 159 and the reported high molecular weight complexes in tumor cells. Neither feature alone establishes the identity of an unexpected band. The supplied record provides no observed band size for comparison.
UniProt reports TM4SF1 in high molecular weight complexes in tumor cells and notes interaction with SDCBP2. This makes a complex worth considering, but these features do not identify any particular Western-blot band.
Consider the annotated N-linked sites at UniProt positions 129 and 159 and the reported high molecular weight complexes in tumor cells. Neither feature alone establishes the identity of an unexpected band. The supplied record provides no observed band size for comparison.
Boster reagents
TM4SF1 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-TM4SF1 Antibody
Cat # A10368
The catalog reports one anti-TM4SF1 antibody, A10368, with stated human, mouse, and rat reactivity. Its Western blot image uses human lung tissue lysate at 0.5 and 1 μg/mL; the supplied evidence does not show mouse or rat WB samples.
Which to pick: A10368 is the only listed option. Choose it when its stated reactivity fits your sample; the supplied WB image documents testing in human lung tissue lysate at 0.5 and 1 μg/mL.
Source: BosterBio TM4SF1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P30408.
- Human Protein Atlas. TM4SF1 tissue expression.
- PMC12391266 — target-verified WB comparison
- PMC5377454 — target-verified WB comparison
- PMC11208120 — target-verified WB comparison