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ACVR1B / Activin receptor type-1B · Western blot design guide
Design a Western Blot for ACVR1B
Source-linked ACVR1B Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ACVR1B 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
ACVR1B 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 | ~56.8 kDa | |
| Observed band | ~52 kDa | |
| Gel | 10% (catalog M02882) | |
| Positive control | Appendix (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Phosphorylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 5 isoform(s) |
Section 1
Source-Linked ACVR1B Western Blot Protocol Options
The M02882 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Caco-2, human MCF-7, human 293T, human SH-SY5Y, rat brain, rat lung, mouse brain, mouse lung (catalog M02882) |
| Gel % | 10% (catalog M02882) |
| Load | 30 ug; reducing conditions (catalog M02882) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M02882) |
| Membrane | nitrocellulose membrane (catalog M02882) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M02882) |
| Primary antibody | M02882 · 1:500 (catalog M02882) |
| Primary incubation | overnight at 4°C (catalog M02882) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog M02882) |
| Secondary incubation | 1.5 hour at RT (catalog M02882) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M02882) |
| Detection | ECL (catalog M02882) |
Section 2
What Is the Expected ACVR1B Western Blot Band Size?
ACVR1B is predicted at 56.8 kDa and observed near 52 kDa; the cause of that difference is not established.
| Band near 52 kDa | Empirical ACVR1B band in reducing lysates; confirm identity with controls |
| Band near 56.8 kDa | Near the predicted full-length precursor mass |
| Band below the precursor position | Could reflect signal-peptide removal; the resulting migration is unverified |
| Band above the precursor position | Could reflect glycosylation at Asn43 if the site is occupied |
| Several bands | Could include isoforms 1 through 5; their migration differences are unknown |
💡Expected ACVR1B appearanceACVR1B has a predicted full-length mass of 56.8 kDa and an empirical band near 52 kDa in reducing lysates; the difference is unexplained, so confirm band identity with ordinary controls.
| 56.8 kDa predicted full-length mass | Reference mass for the precursor; the observed band is near 52 kDa |
| Signal peptide at residues 1–23 | Cleavage can make the mature protein smaller than the precursor; its apparent size is unknown |
| N-linked glycosylation site at Asn43 | Occupancy could increase apparent size; a visible shift is unproven |
| Splice isoforms 1, 2, 3, 4 and 5 | May differ in size; their masses and separation on a blot are unknown |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated ACVR1B may be under-recovered | Check membrane extraction and test an enriched membrane fraction |
| Band higher than expected | Occupancy of the Asn43 glycosylation site is possible | Compare treated and untreated samples and confirm band identity |
| Band lower than expected | The empirical 52 kDa band lies below the 56.8 kDa prediction; the cause is unknown | Check with an independent antibody or ACVR1B depletion control |
| Multiple bands | Isoforms 1 through 5 exist, but distinct migration is unverified | Use isoform-aware controls and verify each band's identity |
| Broad smear instead of sharp band | Variable glycosylation at Asn43 is possible but unverified | Compare deglycosylated and untreated samples and verify the signal |
Experiment evidence
Western blot images showing ACVR1B band
M02882 · ACVR1B
M02882 · ACVR1B
A02882 · ACVR1B
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ACVR1B Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ACVR1B in Western blot, you can use appendix tissue lysate.
Positive control: Appendix (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 ACVR1B is a membrane protein, a membrane-enriched lysate may improve detection.
HPA tissue expression evidence for ACVR1B
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 | Medium | Protein (IHC) | HPA → |
| Breast | adipocytes | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Colon | glandular cells | Medium | Protein (IHC) | HPA → |
| Duodenum | glandular cells | Medium | 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 → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | endothelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ACVR1B Western Blot Tips
Deeper troubleshooting and optimisation questions for ACVR1B, answered from its protein features.
How should ACVR1B 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 ACVR1B isoforms produce different bands?
Isoforms · Five isoforms are listed. Relative to the canonical sequence, isoform 5 lacks residues 1–52; isoform 4 changes residue 271; and isoforms 2 and 3 have altered C-terminal sequences. Check whether the antibody epitope is present in the isoform being measured. The listed sequence changes do not establish where each isoform migrates.
How might glycosylation affect ACVR1B band interpretation?
PTM · UniProt lists one N-linked glycosylation site at Asn43 in the canonical sequence. Isoform 5 lacks residues 1–52, including that site. A migration difference could be investigated in light of this feature, but the site alone does not establish a visible shift or identify a particular band.
UniProt lists phosphotyrosine at residue 380. Check that a phospho-specific antibody targets this residue and that its stated numbering matches UniProt's canonical sequence; antibody or paper numbering may differ. This feature supports site-specific interpretation, but does not establish when phosphorylation occurs or predict a visible band shift.
UniProt lists phosphotyrosine at residue 380. Check that a phospho-specific antibody targets this residue and that its stated numbering matches UniProt's canonical sequence; antibody or paper numbering may differ. This feature supports site-specific interpretation, but does not establish when phosphorylation occurs or predict a visible band shift.
Does this guide establish induction of ACVR1B?
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 ACVR1B Western blot?
Transfer · ACVR1B is a single-pass cell-membrane protein with a predicted mass of 56.8 kDa. Choose and verify a transfer setup that retains signal for a membrane protein in this mass range, using a total-protein stain or another transfer check. The supplied features do not identify a specific transfer method or settings.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M02882 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 ACVR1B bands be quantified across samples?
Quantitation · Use an antibody whose epitope covers the isoforms you intend to quantify, and compare the same verified band across samples. ACVR1B has five listed isoforms, one N-linked glycosylation site, and phosphotyrosine at residue 380; these features can complicate assigning signal to total ACVR1B versus a particular form.
Does the ~52 kDa band match ACVR1B's predicted mass?
Interpretation · ACVR1B is predicted at 56.8 kDa, while the observed band is about 52 kDa. Its signal peptide spans residues 1–23, and it has an N-linked glycosylation site at Asn43. These features may affect migration, but their presence alone does not explain the difference. Confirm band identity before assigning it to ACVR1B.
Check antibody epitope coverage against the listed sequence variants: isoform 5 lacks residues 1–52, isoform 4 changes residue 271, and isoforms 2 and 3 have altered C-terminal sequences. Also consider the signal peptide at 1–23 and N-linked site at Asn43. None of these features alone identifies an unexpected band or proves its migration pattern.
Check antibody epitope coverage against the listed sequence variants: isoform 5 lacks residues 1–52, isoform 4 changes residue 271, and isoforms 2 and 3 have altered C-terminal sequences. Also consider the signal peptide at 1–23 and N-linked site at Asn43. None of these features alone identifies an unexpected band or proves its migration pattern.
Boster reagents
ACVR1B 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-ACVR1B Rabbit Monoclonal Antibody
Cat # M02882
Real WB data
Anti-Activin receptor type-1B ACVR1B Antibody
Cat # A02882
Two anti-ACVR1B antibodies have supplied Western blot images. M02882 shows a band near 52 kDa versus an expected 57 kDa in specified human, rat, and mouse lysates; A02882 shows human kidney lysate with a blocking-peptide comparison. No publication evidence is supplied.
Which to pick: Choose M02882 for the documented human cell, rat tissue, or mouse tissue blot contexts; it lists human, mouse, and rat reactivity. Choose A02882 if the human kidney blot with blocking peptide is most relevant; it lists human and mouse reactivity.
Source: BosterBio ACVR1B gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P36896.
- Human Protein Atlas. ACVR1B tissue expression.
- PMC6912306 — target-verified WB comparison
- PMC13615627 — target-verified WB comparison
- PMC4047430 — target-verified WB comparison