This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
FOLR1 / Folate receptor alpha · Western blot design guide
Design a Western Blot for FOLR1
Source-linked FOLR1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-FOLR1 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
FOLR1 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 | ~29.8 kDa | |
| Gel | 12–15% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Cleaved | |
| Caveat | Glycosylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked FOLR1 Western Blot Protocol Options
The M02254 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | HEK293 cell lysate (catalog M02254) |
| 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 | M02254; use the WB datasheet starting dilution (standard starting point) |
| 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 FOLR1 Western Blot Band Size?
FOLR1 has a predicted 29.8 kDa precursor; cleavage and N-linked glycosylation could alter migration, but no empirical band size or demonstrated shift is supplied.
| Band near 29.8 kDa | Consistent with the predicted full-length precursor; identity needs confirmation |
| Band below 29.8 kDa | Could reflect signal-peptide or propeptide cleavage |
| Band above 29.8 kDa | Could reflect N-linked glycosylation at Asn69, Asn161, or Asn201 |
| Weak or absent lysate band | May reflect enrichment of GPI-anchored FOLR1 at the cell surface |
💡Expected FOLR1 appearanceUniProt predicts a 29.8 kDa precursor; cleavage and N-linked glycosylation may alter migration, but no empirical band size is supplied, so confirm any candidate band with ordinary identity controls.
| Predicted precursor mass | Provides a 29.8 kDa sequence-based reference |
| N-linked glycosylation at Asn69 | May increase apparent mass |
| N-linked glycosylation at Asn161 | May increase apparent mass |
| N-linked glycosylation at Asn201 | May increase apparent mass |
| Signal peptide at residues 1–24 | Cleavage makes the mature chain smaller than the precursor |
| Propeptide at residues 235–257 | Cleavage makes the mature chain smaller than the precursor |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | GPI-anchored FOLR1 is associated with the cell membrane | Check a membrane-enriched fraction and a FOLR1-positive control |
| Band higher than expected | N-linked glycosylation may affect migration | Compare matched samples before and after N-glycan removal |
| Band lower than expected | Signal-peptide or propeptide cleavage may reduce chain size | Confirm identity with a FOLR1-positive control and an independent antibody |
| Broad smear instead of sharp band | Variable N-linked glycosylation is possible at three listed sites | Compare matched samples before and after N-glycan removal |
| Multiple bands | Processing or glycosylation may produce different forms, though distinct bands are unproven | Check which bands persist with an independent FOLR1 antibody and after N-glycan removal |
Experiment evidence
Western blot images showing FOLR1 band
M02254 · FOLR1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for FOLR1 Western blot
🧪For positive controls for FOLR1 in Western blot, you can use an HPA-positive sample once identified; none is supplied here.
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: FOLR1 is GPI-anchored at the cell membrane, and the supplied HPA data cannot establish tissue controls.
HPA tissue expression evidence for FOLR1
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 FOLR1 Western Blot Tips
Deeper troubleshooting and optimisation questions for FOLR1, answered from its protein features.
How should FOLR1 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 annotated FOLR1 isoforms explain multiple bands?
Isoforms · Only one isoform is supplied, with no alternative sequence. Do not assign extra bands to a specific splice isoform from these records. Assess them in light of the annotated processing and glycosylation sites.
Which FOLR1 glycosylation sites matter when assessing band patterns?
PTM · UniProt lists N-linked glycosylation at Asn69, Asn161, and Asn201. Use these UniProt precursor coordinates when comparing annotations; paper or antibody numbering may differ. The site annotations alone do not prove that a particular band or shift is glycosylation-dependent.
Does this guide establish induction of FOLR1?
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 FOLR1?
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 M02254 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 FOLR1 bands be quantified across samples?
Quantitation · Use the same sample fraction and processing conditions across samples. FOLR1 is membrane-associated and has three annotated N-linked glycosylation sites, so record which bands are included in the measurement. These features alone cannot establish that every detected band represents FOLR1.
Why might FOLR1 migrate differently from its predicted 29.8 kDa?
Interpretation · The 29.8 kDa prediction is for the precursor. UniProt annotates a signal peptide at 1..24, a propeptide at 235..257, and three N-linked glycosylation sites. These features can affect the mature protein’s mass or migration, but they do not establish an apparent band size. No observed band size was supplied.
UniProt annotates a signal peptide at 1..24 and a propeptide at 235..257. Consider precursor processing when comparing a band with the 29.8 kDa predicted precursor mass. The supplied features do not give an observed size for a processed form.
UniProt places FOLR1 at the cell membrane, including the apical membrane, and identifies it as GPI-anchored. Include a membrane-containing fraction when assessing detection. Keep fraction preparation consistent when comparing samples.
Check the annotated signal peptide (1..24), propeptide (235..257), and N-linked sites (Asn69, Asn161, Asn201), alongside FOLR1’s membrane and GPI-anchor annotations. Eight disulfide bonds are also listed. None of these features alone identifies an unexpected band or demonstrates a visible shift.
UniProt annotates a signal peptide at 1..24 and a propeptide at 235..257. Consider precursor processing when comparing a band with the 29.8 kDa predicted precursor mass. The supplied features do not give an observed size for a processed form.
UniProt places FOLR1 at the cell membrane, including the apical membrane, and identifies it as GPI-anchored. Include a membrane-containing fraction when assessing detection. Keep fraction preparation consistent when comparing samples.
Check the annotated signal peptide (1..24), propeptide (235..257), and N-linked sites (Asn69, Asn161, Asn201), alongside FOLR1’s membrane and GPI-anchor annotations. Eight disulfide bonds are also listed. None of these features alone identifies an unexpected band or demonstrates a visible shift.
Boster reagents
FOLR1 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-FOLR1 Rabbit Monoclonal Antibody
Cat # M02254
The catalog reports one anti-FOLR1 antibody for Western blot: rabbit monoclonal M02254. Its catalog reports human, mouse, and rat reactivity and shows a blot of FOLR1 expression in HEK293 cell lysate. The supplied evidence shows no other tested specimens.
Which to pick: M02254 is the only listed option. Its WB image uses HEK293 cell lysate; human, mouse, and rat are listed as reactive, but the supplied image does not demonstrate performance in mouse or rat samples.
Source: BosterBio FOLR1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P15328.
- Human Protein Atlas. FOLR1 tissue expression.
- PMC13162693 — target-verified WB comparison
- PMC7930947 — target-verified WB comparison
- PMC5991383 — target-verified WB comparison