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- Table of Contents
MPL / Thrombopoietin receptor · Western blot design guide
Design a Western Blot for MPL
Real validated MPL Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MPL 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
MPL Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~71.2 kDa | |
| Observed band | ~90 kDa | |
| Gel | 10% (catalog A00483-2) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Phosphorylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated MPL Western Blot Protocols
The A00483-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human HEL, human Caco-2 (catalog A00483-2) |
| Gel % | 10% (catalog A00483-2) |
| Load | 30 ug; reducing conditions (catalog A00483-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A00483-2) |
| Membrane | nitrocellulose membrane (catalog A00483-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A00483-2) |
| Primary antibody | A00483-2 · 0.5 μg/mL (catalog A00483-2) |
| Primary incubation | overnight at 4°C (catalog A00483-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A00483-2) |
| Secondary incubation | 1.5 hour at RT (catalog A00483-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A00483-2) |
| Detection | ECL (catalog A00483-2) |
Section 2
What Is the Expected MPL Western Blot Band Size?
MPL is predicted at 71.2 kDa and observed near 90 kDa on a reducing blot; the cause of the difference is not established.
| Band near 90 kDa | Empirical MPL band observed in reducing whole-cell lysates; confirm identity with antibody controls |
| Band near 71 kDa | Near the predicted full-length mass; identity requires confirmation |
| Band below the precursor position | Could reflect removal of the 1–25 signal peptide; size of the mature band is not established |
| Band near twice the monomer position | Could reflect an MPL homodimer that persists during electrophoresis |
| Broad or multiple bands | Could reflect glycoform variation or isoforms 1 and 2; distinct migration is not established |
💡Expected MPL appearanceMPL has a predicted full-length mass of 71.2 kDa, while a reducing whole-cell blot shows an empirical band near 90 kDa; its identity warrants ordinary antibody controls, and the cause of the difference is unestablished.
| Predicted full-length mass | 71.2 kDa by sequence; the empirical reducing-blot band is near 90 kDa |
| N-linked glycosylation at Asn117, Asn178, Asn298 and Asn358 | May affect apparent migration; the size of any shift is not supplied |
| MPL homodimer | May yield a band near twice the monomer position if it persists during electrophoresis |
| Isoforms 1 and 2 | May differ in size; their masses and separation on a blot are not supplied |
| Signal peptide at residues 1–25 | Its removal makes the mature chain smaller than the full-length precursor; the migration difference is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | MPL is a membrane protein and may be poorly recovered in the prepared lysate | Check membrane-protein recovery and use a documented positive lysate |
| Band higher than expected | The near-90 kDa empirical band exceeds the 71.2 kDa sequence prediction; its cause is unestablished | Compare with a positive control and verify antibody specificity; assess reduction if a much higher band appears |
| Band lower than expected | Signal-peptide removal or a different isoform is possible, but neither band position is supplied | Confirm identity with a second antibody or targeted depletion |
| Broad smear instead of sharp band | Heterogeneous occupancy or processing at the four N-linked glycosylation sites is possible | Compare treated and untreated samples after N-glycan removal and confirm the signal with an antibody control |
| Multiple bands | Isoforms 1 and 2 or persistent homodimer may contribute; distinct bands are not established | Compare reducing conditions and confirm individual bands with an independent antibody |
| Weak or no signal | Low MPL abundance or poor recovery of membrane protein | Check loading and transfer, enrich the membrane fraction, and include a documented positive lysate |
Experiment evidence
Western blot images showing MPL band
A00483-2 · MPL
A00483 · MPL
M00483 · MPL
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MPL Western blot
🧪For positive controls for MPL in Western blot, you can use no tissue or cell line identified by the supplied HPA evidence.
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: MPL is membrane localized, and the supplied HPA data cannot establish suitable tissue controls.
HPA tissue expression evidence for MPL
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 MPL Western Blot Tips
Deeper troubleshooting and optimisation questions for MPL, answered from its protein features.
What should be checked when unexpected MPL bands appear?
Band shift · Check antibody epitope coverage against both isoforms and consider the four listed N-linked glycosylation sites when investigating migration. MPL is also reported as a homodimer, but that feature alone does not establish that a higher Western-blot band is a dimer. Band position alone cannot identify an unexpected species.
Could MPL isoforms produce different bands?
Isoforms · UniProt lists two isoforms. Relative to isoform 1, isoform 2 replaces residues 523–579 and lacks residues 580–635. Check whether the antibody recognizes a region retained in both isoforms before assigning a band to either one. The sequence differences alone do not establish their apparent band positions.
How should MPL phosphotyrosine bands be interpreted?
PTM · UniProt lists phosphotyrosines at positions 591, 626, and 631 in canonical isoform 1 numbering. All three positions fall within the region missing from isoform 2. Confirm which isoform and epitope a phospho-specific antibody detects; do not interpret phosphotyrosine signal as total MPL abundance.
Does this guide establish induction of MPL?
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 MPL Western blot?
Transfer · MPL is a single-pass membrane protein, and the supplied observed band is approximately 90 kDa. Verify that your transfer conditions recover the band at that position, for example by checking protein remaining in the gel and protein on the membrane. The supplied UniProt features do not specify a transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A00483-2 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.
How can MPL bands be quantified consistently?
Quantitation · Quantify the same defined band region across samples and document whether it captures the approximately 90 kDa observed band. Keep total MPL and phosphotyrosine measurements separate: UniProt lists three phosphotyrosines, while isoform 2 lacks their canonical positions. Check antibody epitope coverage before comparing isoforms.
Why might MPL migrate near 90 kDa instead of 71.2 kDa?
Interpretation · The observed band is approximately 90 kDa, while the predicted mass is 71.2 kDa. UniProt lists N-linked glycosylation at Asn117, Asn178, Asn298, and Asn358, plus a signal peptide at residues 1–25. These features are relevant when assessing migration, but they do not establish the cause or size of the difference.
Boster reagents
MPL 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-TPOR/MPL Antibody Picoband®
Cat # A00483-2
Real WB data
Anti-CD110 MPL Antibody
Cat # A00483
Real WB data
Anti-TPOR Rabbit Monoclonal Antibody
Cat # M00483
Three the supplier anti-MPL antibodies have WB images. A00483-2 shows a band near 90 kDa in human HEL and Caco-2 lysates, versus an expected 70 kDa. A00483 shows MCF7 cells; M00483 shows A431 and NIH/3T3 lysates. No publication evidence is supplied.
Which to pick: For human HEL or Caco-2 lysates, A00483-2 has a detailed WB caption but lists human reactivity only. A00483 and M00483 list human, mouse and rat reactivity; M00483 shows NIH/3T3 lysate, while A00483 shows MCF7 cells. All three have WB images.
Source: BosterBio MPL gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P40238.
- Human Protein Atlas. MPL tissue expression.
- PMC549047 — target-verified WB comparison
- PMC13408313 — target-verified WB comparison