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- Table of Contents
BMP5 / Bone morphogenetic protein 5 · Western blot design guide
Design a Western Blot for BMP5
Source-linked BMP5 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-BMP5 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
BMP5 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 | ~51.7 kDa | |
| Observed band | ~51 kDa | |
| Gel | 5–20% (catalog PB9689) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Cleaved | |
| Caveat | Processing-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked BMP5 Western Blot Protocol Options
The PB9689 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Rat Liver at 50ug, Mouse Liver at 50ug, A549 at 40ug (catalog PB9689) |
| Gel % | 5–20% (catalog PB9689) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog PB9689) |
| Membrane | nitrocellulose membrane (catalog PB9689) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog PB9689) |
| Primary antibody | PB9689 · 0.5 μg/mL (catalog PB9689) |
| Primary incubation | overnight at 4°C (catalog PB9689) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog PB9689) |
| Secondary incubation | 1.5 hour at RT (catalog PB9689) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog PB9689) |
| Detection | ECL (catalog PB9689) |
Section 2
What Is the Expected BMP5 Western Blot Band Size?
BMP5 has a predicted precursor mass of 51.7 kDa and an observed band near 51 kDa; the cause of their difference is not established.
| Band near 51 kDa | Empirical BMP5 band in antibody QC; confirm identity with controls |
| Band below 51 kDa | Could reflect signal peptide or propeptide cleavage |
| Band near twice a monomer band | Could reflect a disulfide-linked BMP5 dimer under incomplete reduction |
| Bands with differing mobility | Could reflect BMP5 glycosylation; the listed sites do not establish a visible shift |
| Several discrete bands | Could reflect isoforms 1 and 2 or processing; distinct isoform bands are unverified |
| Little or no band in whole-cell lysate | BMP5 is secreted |
💡Expected BMP5 appearanceUniProt predicts 51.7 kDa for the BMP5 precursor, while antibody QC reports a band near 51 kDa; confirm band identity with appropriate controls because the supplied features do not establish its molecular state.
| UniProt precursor mass | Predicted mass is 51.7 kDa; antibody QC observes a band near 51 kDa |
| N-linked glycosylation sites at Asn211, Asn327, Asn345, and Asn395 | Glycosylation could alter apparent size, but site annotations do not establish a visible shift |
| Disulfide linkage at Cys418 | A linked dimer could migrate near twice its monomer size if reduction is incomplete |
| Signal peptide at residues 1–30 | Cleavage could make the processed protein smaller than the precursor |
| Propeptide at residues 31–316 | Cleavage could produce a smaller mature protein |
| Splice isoforms 1 and 2 | Their sizes may differ, but distinct migration is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | BMP5 is secreted | Check conditioned medium alongside lysate |
| Band higher than expected | Incomplete reduction could retain a disulfide-linked species | Compare fully reducing and nonreducing preparations |
| Band lower than expected | Signal peptide or propeptide cleavage could produce a smaller form | Check antibody epitope and compare precursor and mature-form controls |
| Broad smear instead of sharp band | Variable N-linked glycosylation is possible | Compare untreated and deglycosylated samples |
| Multiple bands | Processing, glycosylation, or isoforms 1 and 2 could contribute | Compare reduction and deglycosylation conditions and verify band identity with a BMP5 control |
| Fragments below expected size | BMP5 has a cleavable propeptide | Check antibody epitope and use a BMP5 identity control before assigning fragments |
Experiment evidence
Western blot images showing BMP5 band
PB9689 · BMP5
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for BMP5 Western blot
🧪For positive controls for BMP5 in Western blot, you can use a separately validated BMP5-positive sample because HPA provides no positive tissue or cell candidate.
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.
⚠️Feasibility: BMP5 is secreted, so conditioned medium may give a stronger signal than whole-cell lysate.
HPA tissue expression evidence for BMP5
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 BMP5 Western Blot Tips
Deeper troubleshooting and optimisation questions for BMP5, answered from its protein features.
How should BMP5 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 BMP5 isoform 2 differ on a blot?
Isoforms · UniProt lists isoforms 1 and 2. Isoform 2 lacks residues 369..405 in full-length sequence numbering, a segment containing the listed glycosylation site at 395. It could therefore differ in migration or antibody recognition, depending on the antibody epitope; the features alone do not establish a specific band position.
No conclusion about isoform identity follows from mass alone. The ~51 kDa observed band is near the 51.7 kDa full-length prediction, while isoform 2 lacks residues 369..405 and BMP5 has processing and glycosylation features. Interpret the band alongside the antibody's recognized region and the sample type.
No conclusion about isoform identity follows from mass alone. The ~51 kDa observed band is near the 51.7 kDa full-length prediction, while isoform 2 lacks residues 369..405 and BMP5 has processing and glycosylation features. Interpret the band alongside the antibody's recognized region and the sample type.
Could glycosylation affect BMP5 band position?
PTM · UniProt lists N-linked glycosylation at positions 211, 327, 345 and 395, using full-length sequence numbering. Position 211 lies in the annotated propeptide; the other three lie beyond it. These sites make glycosylation relevant to interpretation, but their presence alone does not predict a visible shift.
Does this guide establish induction of BMP5?
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 BMP5?
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 PB9689 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 secreted BMP5 be sampled for quantitation?
Quantitation · BMP5 is annotated as secreted. Specify whether measurements come from conditioned medium or cell lysate, and compare like sample types. The signal peptide, propeptide and glycosylation features also make it useful to define which band is being quantified.
Does the ~51 kDa band represent mature BMP5?
Interpretation · The predicted 51.7 kDa mass is for the full-length sequence, while UniProt lists a signal peptide at 1..30 and a propeptide at 31..316. An observed ~51 kDa band is close to the full-length prediction, but apparent mass alone cannot establish whether BMP5 has been processed.
BMP5 has a signal peptide at 1..30 and a propeptide at 31..316. Consider signal peptide removal and propeptide cleavage when interpreting bands of different sizes. These features do not establish which processed forms are present in a sample.
UniProt lists four disulfide bonds. Compare reducing and nonreducing preparations if bands differ between conditions, and record the condition used for quantitation. The bond count alone cannot identify an unexpected band or predict its apparent mass.
BMP5 has a signal peptide at 1..30 and a propeptide at 31..316. Consider signal peptide removal and propeptide cleavage when interpreting bands of different sizes. These features do not establish which processed forms are present in a sample.
UniProt lists four disulfide bonds. Compare reducing and nonreducing preparations if bands differ between conditions, and record the condition used for quantitation. The bond count alone cannot identify an unexpected band or predict its apparent mass.
Boster reagents
BMP5 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-BMP5 Antibody Picoband®
Cat # PB9689
PB9689 is the listed anti-BMP5 antibody, with stated Human, Mouse, and Rat reactivity. Its Western blot image shows an approximately 51 kDa band in rat and mouse liver lysates and A549 cell lysate. No independent validation evidence is supplied.
Which to pick: PB9689 is the only listed option and has a Western blot image. Its caption reports bands in rat and mouse liver lysates and A549 cell lysate; use those tested samples and the reported 51 kDa band to guide your choice.
Source: BosterBio BMP5 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P22003.
- Human Protein Atlas. BMP5 tissue expression.
- PMC13357077 — target-verified WB comparison
- PMC10265437 — target-verified WB comparison