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MFSD2A / Sodium-dependent lysophosphatidylcholine symporter 1 · Western blot design guide
Design a Western Blot for MFSD2A
Source-linked MFSD2A Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MFSD2A 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
MFSD2A 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 | ~60.2 kDa | |
| Observed band | ~60 kDa | |
| Gel | 10% (catalog A01967-2) | |
| Positive control | Appendix (IHC candidate; verify WB) +4 more | |
| Negative control | Heart muscle (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 3 isoform(s) |
Section 1
Source-Linked MFSD2A Western Blot Protocol Options
The A01967-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human THP-1, rat liver, mouse liver (catalog A01967-2) |
| Gel % | 10% (catalog A01967-2) |
| Load | 30 ug; reducing conditions (catalog A01967-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A01967-2) |
| Membrane | nitrocellulose membrane (catalog A01967-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A01967-2) |
| Primary antibody | A01967-2 · 0.5 μg/mL (catalog A01967-2) |
| Primary incubation | overnight at 4°C (catalog A01967-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A01967-2) |
| Secondary incubation | 1.5 hour at RT (catalog A01967-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A01967-2) |
| Detection | ECL (catalog A01967-2) |
Section 2
What Is the Expected MFSD2A Western Blot Band Size?
MFSD2A is predicted at 60.2 kDa and observed at ~60 kDa; the cause of any small apparent difference is not established.
| Band at ~60 kDa | Matches the observed MFSD2A band and its 60.2 kDa predicted mass; confirm identity with an independent antibody or target depletion. |
| Band above ~60 kDa | N-linked glycosylation at Asn230 or Asn240 could affect migration, but a visible shift is not established. |
| Multiple bands | Isoforms 1, 2, and 3 may differ in size, though distinct bands are not established. |
| Weak or absent band in a soluble fraction | MFSD2A resides in cell and endoplasmic reticulum membranes. |
💡Expected MFSD2A appearanceMFSD2A is predicted at 60.2 kDa and observed at ~60 kDa in reducing blots; confirm band identity with an independent antibody or target depletion.
| UniProt predicted mass | Places the reference protein near 60.2 kDa. |
| N-linked glycosylation at Asn230 | Could alter apparent size; a measurable shift is not established. |
| N-linked glycosylation at Asn240 | Could alter apparent size; a measurable shift is not established. |
| Splice isoforms 1, 2, and 3 | May differ in size, but their individual masses and migration are not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated MFSD2A may be poorly extracted. | Check membrane solubilization and a membrane fraction; include a positive lysate. |
| Band higher than expected | N-linked glycosylation at Asn230 or Asn240 could affect migration. | Compare untreated and deglycosylated samples and verify band identity independently. |
| Band lower than expected | An isoform may differ in size, but isoform masses are unknown. | Check isoform expression and verify the band with an independent antibody or target depletion. |
| Multiple bands | Isoforms or differing glycosylation at Asn230 and Asn240 are possible. | Compare deglycosylated samples and use an independent band-identity control. |
| Weak or no signal | Extraction of this multi-pass membrane protein may be inefficient. | Optimize membrane protein recovery and check sample loading. |
Experiment evidence
Western blot images showing MFSD2A band
A01967-2 · MFSD2A
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MFSD2A Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for MFSD2A in Western blot, you can use appendix tissue lysate, which HPA scores High.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Heart muscle (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, MFSD2A may require membrane-enriched lysate for a clear signal.
HPA tissue expression evidence for MFSD2A
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 |
|---|---|---|---|---|
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Salivary gland | glandular cells | High | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Prostate | glandular cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MFSD2A Western Blot Tips
Deeper troubleshooting and optimisation questions for MFSD2A, answered from its protein features.
How should MFSD2A 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 MFSD2A isoforms produce different bands?
Isoforms · Three isoforms are listed. Relative to the canonical sequence, isoform 2 lacks residues 77–89, while isoform 3 replaces residues 1–186 with a different sequence. Check which isoform an antibody recognizes before assigning bands; these sequence differences alone do not establish their apparent positions.
Which glycosylation sites matter when interpreting MFSD2A bands?
PTM · UniProt annotates N-linked glycosylation at asparagines 230 and 240, using canonical UniProt numbering. Keep that numbering explicit when comparing antibody or paper annotations. The annotations identify potential modifications; they do not establish how much either site changes migration.
Does this guide establish induction of MFSD2A?
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 MFSD2A Western blot?
Transfer · MFSD2A is a 543-residue, multi-pass membrane protein with a predicted mass of 60.2 kDa. Choose conditions appropriate for transferring a membrane protein near 60 kDa, then check transfer efficiency. The supplied features do not identify a specific membrane, buffer, or transfer time.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A01967-2 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.
What should be checked when quantifying an MFSD2A band?
Quantitation · Use a consistent band definition across samples and verify that the signal falls within the assay’s quantitative range. MFSD2A is annotated at the cell and endoplasmic reticulum membranes, so keep the sampled fraction consistent. If multiple bands appear, establish which represents the isoform or form being measured before combining signals.
How should the ~60 kDa band compare with MFSD2A’s predicted mass?
Interpretation · The observed band is approximately 60 kDa, close to the predicted 60.2 kDa. MFSD2A has two annotated N-linked glycosylation sites, but their presence alone does not establish a visible shift or explain any difference between apparent and predicted mass.
Consider antibody recognition of the listed isoforms: isoform 2 lacks canonical residues 77–89, and isoform 3 has a different sequence at residues 1–186. Compare the antibody’s target region with those changes. The supplied features do not establish that either isoform produces a particular lower band.
MFSD2A is annotated with one disulfide bond and an interaction with ERVFRD-1/syncytin-2, but neither annotation identifies a higher Western-blot band. Its N-linked sites at canonical positions 230 and 240 also do not establish a visible shift. Treat a higher band as unassigned until its identity is verified.
Consider antibody recognition of the listed isoforms: isoform 2 lacks canonical residues 77–89, and isoform 3 has a different sequence at residues 1–186. Compare the antibody’s target region with those changes. The supplied features do not establish that either isoform produces a particular lower band.
MFSD2A is annotated with one disulfide bond and an interaction with ERVFRD-1/syncytin-2, but neither annotation identifies a higher Western-blot band. Its N-linked sites at canonical positions 230 and 240 also do not establish a visible shift. Treat a higher band as unassigned until its identity is verified.
Boster reagents
MFSD2A 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-MFSD2A Antibody Picoband®
Cat # A01967-2
A01967-2 is a rabbit polyclonal anti-MFSD2A antibody with a Western blot image showing an approximately 60 kDa band in human THP-1 cell, rat liver, and mouse liver lysates. The supplied evidence is limited to this product example.
Which to pick: A01967-2 is the only listed option. It reports human, mouse, and rat reactivity and has a Western blot image for the named lysates. Choose it when those reported contexts fit your experiment; other samples require your own validation.
Source: BosterBio MFSD2A gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q8NA29.
- Human Protein Atlas. MFSD2A tissue expression.
- PMC7038791 — target-verified WB comparison
- PMC5586300 — target-verified WB comparison
- PMC11726956 — target-verified WB comparison