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TNFSF15 / Tumor necrosis factor ligand superfamily member 15 · Western blot design guide
Design a Western Blot for TNFSF15
Source-linked TNFSF15 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TNFSF15 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
TNFSF15 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 | ~28.1 kDa | |
| Observed band | ~28 kDa | |
| Gel | 5–20% (catalog A02402-1) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Source-Linked TNFSF15 Western Blot Protocol Options
The A02402-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | rat liver, rat kidney, mouse kidney (catalog A02402-1) |
| Gel % | 5–20% (catalog A02402-1) |
| Load | 50ug; reducing conditions (catalog A02402-1) |
| Transfer | a Nitrocellulose membrane at 150mA for 50-90 minutes (catalog A02402-1) |
| Membrane | Nitrocellulose membrane (catalog A02402-1) |
| Blocking | 5% Non-fat Milk/ TBS for 1.5 hour at RT (catalog A02402-1) |
| Primary antibody | A02402-1 · 0.5 μg/mL (catalog A02402-1) |
| Primary incubation | overnight at 4°C (catalog A02402-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:10000 (catalog A02402-1) |
| Secondary incubation | 1.5 hour at RT (catalog A02402-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A02402-1) |
| Detection | ECL (catalog A02402-1) |
Section 2
What Is the Expected TNFSF15 Western Blot Band Size?
TNFSF15 is predicted at 28.1 kDa and observed at ~28 kDa; the supplied evidence does not establish a cause for the small difference.
| Band at ~28 kDa | Matches the observed TNFSF15 band and the 28.1 kDa predicted mass |
| Band above ~28 kDa | N-linked glycans at Asn133 or Asn229 could affect migration; the cause requires testing |
| Several bands | Isoforms 1, 2, and 3 are annotated, but distinct band sizes are not established |
| Weak or absent band in soluble lysate | Membrane-associated TNFSF15 may be poorly recovered |
| Mobility change under nonreducing conditions | The intrachain disulfide between residues 162 and 202 could affect migration |
💡Expected TNFSF15 appearanceTNFSF15 has a predicted mass of 28.1 kDa and an observed band at ~28 kDa; confirm band identity with appropriate controls, since the listed features do not establish additional band positions.
| Predicted mass | The 28.1 kDa prediction closely matches the observed ~28 kDa band |
| N-linked glycosylation at Asn133 | Could alter apparent mass if occupied; a visible shift is not established |
| N-linked glycosylation at Asn229 | Could alter apparent mass if occupied; a visible shift is not established |
| Splice isoforms 1, 2, and 3 | Could differ in size, but their individual masses and migration are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated TNFSF15 may be poorly recovered | Check membrane extraction and use a positive control |
| Band higher than expected | N-linked glycosylation is possible; its effect on this band is unproven | Compare untreated and deglycosylated samples and verify band identity |
| Band lower than expected | An isoform is possible, but its migration is unknown | Check isoform coverage and verify band identity with a specific control |
| Broad smear instead of sharp band | Variable occupancy or composition of N-linked glycans is possible | Compare with a deglycosylated sample and verify specificity |
| Multiple bands | Three splice isoforms are annotated, but distinct bands are unconfirmed | Check isoform recognition and verify bands with a specific control |
| Weak or no signal | Membrane-associated protein may be underrepresented in the preparation | Enrich the membrane fraction and check a positive control |
Experiment evidence
Western blot images showing TNFSF15 band
A02402-1 · TNFSF15
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TNFSF15 Western blot
🧪For positive controls for TNFSF15 in Western blot, you can use a validated positive sample once one is identified; the supplied HPA evidence names none.
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 samples.
⚠️Feasibility: TNFSF15 is membrane-associated and secreted, so consider testing both cell lysate and conditioned medium; HPA provides no control samples.
HPA tissue expression evidence for TNFSF15
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 TNFSF15 Western Blot Tips
Deeper troubleshooting and optimisation questions for TNFSF15, answered from its protein features.
How should TNFSF15 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.
How might TNFSF15 isoforms affect band interpretation?
Isoforms · UniProt lists three isoforms. Relative to canonical isoform 1, isoform 2 replaces residues 1–85, while isoform 3 lacks residues 1–77 and replaces residues 78–100. These are canonical sequence coordinates. An antibody's binding region determines which isoforms it can detect; check that region before assigning bands.
Could glycosylation shift the TNFSF15 band?
PTM · UniProt lists N-linked glycosylation at Asn133 and Asn229, using canonical sequence coordinates. These sites make altered migration possible, but their presence alone does not establish a visible shift. Compare matched samples with and without N-glycan removal to assess its contribution.
Does this guide establish induction of TNFSF15?
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 TNFSF15?
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 A02402-1 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 TNFSF15 bands be quantified?
Quantitation · Measure bands consistently across samples prepared under the same fractionation and reducing conditions. Because TNFSF15 has three isoforms and two annotated N-glycosylation sites, define which band or bands are included and use the same rule throughout the comparison.
Why does TNFSF15 appear near 28 kDa?
Interpretation · The observed band near 28 kDa agrees with the predicted 28.1 kDa mass of the canonical protein. Apparent migration alone does not establish which isoform is present.
TNFSF15 is annotated as a single-pass type II membrane protein. Include a membrane-containing fraction when assessing cellular TNFSF15, and keep fraction preparation consistent across samples.
UniProt describes TNFSF15 as a homotrimer and lists one disulfide bond. These features make oligomeric species worth considering, but they do not establish that a higher Western-blot band is a trimer. Compare reducing and nonreducing conditions before making that assignment.
Check whether the antibody can recognize isoforms 1, 2, and 3; whether glycosylation at canonical Asn133 or Asn229 affects migration; and whether reducing conditions change the pattern. These features suggest checks, but none identifies an unexpected band on its own.
TNFSF15 is annotated as a single-pass type II membrane protein. Include a membrane-containing fraction when assessing cellular TNFSF15, and keep fraction preparation consistent across samples.
UniProt describes TNFSF15 as a homotrimer and lists one disulfide bond. These features make oligomeric species worth considering, but they do not establish that a higher Western-blot band is a trimer. Compare reducing and nonreducing conditions before making that assignment.
Check whether the antibody can recognize isoforms 1, 2, and 3; whether glycosylation at canonical Asn133 or Asn229 affects migration; and whether reducing conditions change the pattern. These features suggest checks, but none identifies an unexpected band on its own.
Boster reagents
TNFSF15 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-TL1A/TNFSF15 Antibody Picoband®
Cat # A02402-1
The catalog reports one anti-TNFSF15 antibody for Western blot, A02402-1, with reported Human, Mouse, and Rat reactivity. Its WB image shows an approximately 28 kDa band in rat liver, rat kidney, and mouse kidney lysates; no human WB sample is shown.
Which to pick: A02402-1 is the only listed option. Its WB image supports the reported rat liver, rat kidney, and mouse kidney contexts under the captioned conditions. Human reactivity is listed, but the supplied image does not demonstrate human WB performance.
Source: BosterBio TNFSF15 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O95150.
- Human Protein Atlas. TNFSF15 tissue expression.
- PMC5221472 — target-verified WB comparison
- PMC3485222 — target-verified WB comparison
- PMC4881442 — target-verified WB comparison