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- Table of Contents
TFR2 / Transferrin receptor protein 2 · Western blot design guide
Design a Western Blot for TFR2
Source-linked TFR2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TFR2 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
TFR2 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 | ~88.8 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Liver (IHC candidate; verify WB) | |
| Negative control | Adipose tissue (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 TFR2 Western Blot Protocol Options
The M02353-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | HepG2 cell lysate (catalog M02353-1) |
| Gel % | 8–10% (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 | M02353-1; 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 TFR2 Western Blot Band Size?
The predicted TFR2 monomer is 88.8 kDa; glycans, homodimerization and isoforms may affect migration, but no band position is demonstrated.
| Band near 88.8 kDa | Consistent with the predicted TFR2 monomer; identity requires confirmation. |
| Band above 88.8 kDa | N-linked glycosylation at Asn240, Asn339, Asn540 and Asn754 could affect migration; a shift is not established. |
| Band near 178 kDa under nonreducing conditions | Consistent with a disulfide-linked TFR2 homodimer. |
| Several bands at different sizes | Alpha, Beta and Gamma isoforms are possible contributors; distinct migration is not established. |
💡Expected TFR2 appearanceThe predicted TFR2 monomer is 88.8 kDa; no empirical band size is supplied, so confirm band identity with reduction, deglycosylation and an independent antibody.
| Predicted monomer mass | Sets an 88.8 kDa reference, not a validated band position. |
| N-linked glycosylation at Asn240, Asn339, Asn540 and Asn754 | May alter apparent size; the size of any shift is unknown. |
| Disulfide-linked homodimer | May yield a band near twice the monomer mass without complete reduction. |
| Alpha, Beta and Gamma isoforms | May differ in size; their masses and migration are not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | TFR2 is a single-pass membrane protein and may be poorly recovered. | Check membrane extraction and a membrane-protein control. |
| Band higher than expected | A disulfide-linked homodimer may persist with incomplete reduction; N-linked glycans may also affect migration. | Compare reducing conditions and test deglycosylation. |
| Band lower than expected | An alternative isoform could differ in size, but its migration is unknown. | Check isoform coverage and confirm identity with a second antibody. |
| Broad smear instead of sharp band | Variation in N-linked glycosylation is possible but unproven. | Compare untreated and deglycosylated samples. |
| Multiple bands | Alpha, Beta and Gamma isoforms or different oligomeric states may contribute. | Compare reducing conditions and confirm bands with a second antibody. |
| Weak or no signal | Membrane-protein extraction may be inefficient. | Check lysate preparation and loading with a membrane-protein control. |
Experiment evidence
Western blot images showing TFR2 band
M02353-1 · TFR2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TFR2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TFR2 in Western blot, you can use liver tissue.
Positive control: Liver (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 membrane protein, TFR2 may be easier to detect in membrane-enriched lysates.
HPA tissue expression evidence for TFR2
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 |
|---|---|---|---|---|
| Liver | hepatocytes | High | 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 → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TFR2 Western Blot Tips
Deeper troubleshooting and optimisation questions for TFR2, answered from its protein features.
How should TFR2 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 TFR2 isoforms produce different bands?
Isoforms · UniProt lists Alpha, Beta, and Gamma. Relative to the canonical sequence, Beta lacks residues 1–171 and Gamma lacks residues 343–369. Check whether the antibody epitope is retained in each isoform before assigning bands. These sequence differences do not, by themselves, establish which bands will be visible.
To detect all three listed isoforms, choose an epitope outside canonical residues 1–171 and 343–369. An epitope within either deleted segment may miss the corresponding isoform. Confirm the antibody's stated epitope and numbering convention.
To detect all three listed isoforms, choose an epitope outside canonical residues 1–171 and 343–369. An epitope within either deleted segment may miss the corresponding isoform. Confirm the antibody's stated epitope and numbering convention.
Which TFR2 glycosylation sites should I consider?
PTM · UniProt lists N-linked glycosylation at Asn240, Asn339, Asn540, and Asn754. These are UniProt canonical-sequence coordinates; check the numbering convention before comparing them with an antibody datasheet or publication.
Does this guide establish induction of TFR2?
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 TFR2?
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 M02353-1 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.
What should I control when quantifying TFR2 bands?
Quantitation · TFR2 is a single-pass cell-membrane protein with three listed isoforms. Use the same sample preparation and membrane-protein recovery procedure across samples, and quantify a consistently identified band. If multiple bands appear, avoid combining them until you have established what each represents.
Why might TFR2 migrate differently from its predicted 88.8 kDa mass?
Interpretation · The 88.8 kDa value is calculated from sequence. UniProt lists four N-linked glycosylation sites, but their presence alone does not establish a visible shift or explain any measured band. No observed band size was supplied, so compare bands with an appropriate molecular weight marker.
UniProt describes TFR2 as a homodimer and lists two disulfide bonds. A higher band could be investigated in that context, but these features alone do not identify it as a dimer. Compare samples prepared under consistent reducing conditions before interpreting the band.
UniProt describes TFR2 as a homodimer and lists two disulfide bonds. A higher band could be investigated in that context, but these features alone do not identify it as a dimer. Compare samples prepared under consistent reducing conditions before interpreting the band.
Boster reagents
TFR2 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-Transferrin Receptor 2 Rabbit Monoclonal Antibody
Cat # M02353-1
The catalog reports one anti-TFR2 antibody for Western blotting: M02353-1, a rabbit monoclonal with reported human reactivity. Its WB image shows Transferrin Receptor 2 expression in HepG2 cell lysate; evidence for other samples is not supplied.
Which to pick: M02353-1 is the only listed option. Its HepG2 cell lysate WB image provides a tested context for human TFR2; check suitability separately for other samples.
Source: BosterBio TFR2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9UP52.
- Human Protein Atlas. TFR2 tissue expression.
- PMC11758335 — target-verified WB comparison
- PMC3386558 — target-verified WB comparison
- PMC2746864 — target-verified WB comparison