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- Table of Contents
APOH / Beta-2-glycoprotein 1 · Western blot design guide
Design a Western Blot for APOH
Source-linked APOH Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-APOH 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
APOH 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 | ~38.3 kDa | |
| Observed band | ~50 kDa | |
| Gel | 10% (catalog M01998-1) | |
| 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 Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked APOH Western Blot Protocol Options
The M01998-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human HepG2, human Caco-2, mouse HEPA1-6 (catalog M01998-1) |
| Gel % | 10% (catalog M01998-1) |
| Load | 30 ug; reducing conditions (catalog M01998-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M01998-1) |
| Membrane | nitrocellulose membrane (catalog M01998-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M01998-1) |
| Primary antibody | M01998-1 · 1:1000 (catalog M01998-1) |
| Primary incubation | overnight at 4°C (catalog M01998-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog M01998-1) |
| Secondary incubation | 1.5 hour at RT (catalog M01998-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M01998-1) |
| Detection | ECL (catalog M01998-1) |
Section 2
What Is the Expected APOH Western Blot Band Size?
APOH is predicted at 38.3 kDa, but an empirical reducing-lysate band appears at ~50 kDa; the cause of the difference is not established.
| Band at ~50 kDa | Empirical APOH band reported in reducing whole-cell lysates; confirm identity with antibody controls. |
| Band near 38.3 kDa | Near the predicted mass of the full-length precursor; identity requires confirmation. |
| Band below the precursor position | Could reflect removal of the 1–19 signal peptide; its migration is not established. |
| Diffuse band near the APOH position | Could reflect variable glycosylation at the annotated sites; this pattern is not established. |
| Little or no band in whole-cell lysate | APOH is secreted and may be more readily detected in conditioned medium. |
💡Expected APOH appearanceAPOH has a predicted full-length mass of 38.3 kDa, while antibody QC reports a ~50 kDa band in reducing lysates; the cause of that difference is unestablished, so confirm band identity with antibody controls.
| Predicted full-length mass | 38.3 kDa is the calculated precursor mass, not its demonstrated migration. |
| Signal peptide at residues 1–19 | Cleavage makes the mature chain smaller than the precursor; its band size is not supplied. |
| O-linked GalNAc sites at Thr33 and Thr149 | May affect apparent migration; no visible shift is established for these sites. |
| Complex N-linked glycosylation at Asn162 | May affect apparent migration; its contribution to the observed band is unquantified. |
| N-linked glycosylation at Asn183, Asn193 and Asn253 | May affect apparent migration; no site-specific band shift is established. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | APOH is secreted. | Check conditioned medium alongside lysate and a validated positive control. |
| Band higher than expected | The reported ~50 kDa band exceeds the 38.3 kDa predicted precursor mass; the cause is unestablished. | Compare with the reported reducing-lysate band and verify antibody specificity. |
| Band lower than expected | Signal-peptide cleavage can reduce chain mass, but a lower band is not established here. | Compare lysate and conditioned medium with a validated positive control. |
| Broad smear instead of sharp band | Variation at APOH glycosylation sites is possible but unconfirmed. | Compare untreated and glycosidase-treated samples with appropriate controls. |
| Multiple bands | Different glycosylation or processing states are possible; their bands are not established. | Compare conditioned medium and lysate, and verify bands with an independent APOH antibody. |
| Weak or no signal | Secretion may leave little APOH in the sampled lysate. | Check conditioned medium and include a validated positive control. |
Experiment evidence
Western blot images showing APOH band
M01998-1 · APOH
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for APOH Western blot
🧪For positive controls for APOH in Western blot, you can use no HPA-supported positive sample from the supplied 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: APOH is secreted, so whole-cell lysate may show little signal; conditioned medium may be more suitable.
HPA tissue expression evidence for APOH
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 APOH Western Blot Tips
Deeper troubleshooting and optimisation questions for APOH, answered from its protein features.
How should APOH 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.
Should multiple APOH bands be assigned to isoforms?
Isoforms · The supplied UniProt record lists one isoform and no alternative sequence. Do not assign additional bands to annotated isoforms on this evidence alone.
Which APOH glycosylation sites matter when assessing band shifts?
PTM · UniProt annotates O-linked threonines at 33 and 149, and N-linked asparagines at 162, 183, 193, and 253. These are UniProt sequence coordinates; antibody or paper numbering may differ. The annotations alone do not predict a visible shift.
Define the band region consistently across samples when quantifying the approximately 50 kDa signal. Because APOH has annotated N-linked and O-linked glycosylation sites, assess any differently migrating signal separately before combining it with the main band.
Define the band region consistently across samples when quantifying the approximately 50 kDa signal. Because APOH has annotated N-linked and O-linked glycosylation sites, assess any differently migrating signal separately before combining it with the main band.
Does this guide establish induction of APOH?
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 APOH?
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 M01998-1 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.
How should APOH be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
Why might APOH appear near 50 kDa instead of 38.3 kDa?
Interpretation · The observed band is approximately 50 kDa, while the predicted mass is 38.3 kDa. APOH has six annotated glycosylation sites, which could affect migration, but these features alone do not establish the cause of the difference.
APOH has a signal peptide at UniProt residues 1–19 and is annotated as secreted. Consider that the secreted protein may lack this segment when comparing a band with the 38.3 kDa predicted mass. This feature does not by itself explain the approximately 50 kDa band.
Check them against the approximately 50 kDa observed band and the 38.3 kDa predicted mass. Glycosylation and the 1–19 signal peptide are relevant features, but neither identifies an unexpected band on its own. The record supplies no alternative sequence to support an isoform assignment.
APOH has a signal peptide at UniProt residues 1–19 and is annotated as secreted. Consider that the secreted protein may lack this segment when comparing a band with the 38.3 kDa predicted mass. This feature does not by itself explain the approximately 50 kDa band.
Check them against the approximately 50 kDa observed band and the 38.3 kDa predicted mass. Glycosylation and the 1–19 signal peptide are relevant features, but neither identifies an unexpected band on its own. The record supplies no alternative sequence to support an isoform assignment.
Boster reagents
APOH 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-ApoH Rabbit Monoclonal Antibody
Cat # M01998-1
M01998-1 is an anti-APOH rabbit monoclonal antibody listed for human, mouse, and rat. Its WB image shows a band near 50 kDa in human HepG2 and Caco-2 and mouse HEPA1-6 lysates, although the stated expected size is 38 kDa. Rat WB data are not shown.
Which to pick: M01998-1 is the only listed option and has a WB image for the named human and mouse lysates. Consider the reported 50 kDa band versus the expected 38 kDa when interpreting results; the supplied image does not show rat lysate.
Source: BosterBio APOH gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P02749.
- Human Protein Atlas. APOH tissue expression.
- PMC11445452 — target-verified WB comparison
- PMC8143051 — target-verified WB comparison
- PMC10303272 — target-verified WB comparison