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- Table of Contents
APOD / Apolipoprotein D · Western blot design guide
Design a Western Blot for APOD
Real validated APOD Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-APOD 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
APOD 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 | ~21.3 kDa | |
| Gel | 15% (standard starting point) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Cervix (IHC candidate; verify WB) |
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
Real Curated APOD Western Blot Protocols
The M02196 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Human plasma lysate (catalog M02196) |
| Gel % | 15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.2 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M02196; 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 APOD Western Blot Band Size?
APOD has a predicted 21.3 kDa precursor; processing, glycosylation, and dimerization may affect migration, but no empirical band size is available.
| Band near 21.3 kDa | Compatible with the predicted precursor mass; confirm band identity |
| Band below precursor size | Compatible with cleavage of the 20-residue signal peptide |
| Higher or diffuse band | May reflect complex N-linked glycosylation at Asn65 and Asn98 |
| Band near twice the monomer size | Compatible with a homodimer under non-reducing conditions |
| Little or no band in whole-cell lysate | APOD is secreted |
💡Expected APOD appearanceThe predicted 21.3 kDa mass describes the precursor; signal-peptide cleavage, complex N-linked glycosylation, and dimerization may affect migration, but no empirical band size is supplied, so confirm identity with appropriate controls.
| Predicted precursor mass | 21.3 kDa is the sequence-based reference, not a measured band |
| N-linked glycosylation at Asn65 | Complex glycosylation may increase apparent size |
| N-linked glycosylation at Asn98 | Complex glycosylation may increase apparent size |
| Signal peptide at residues 1–20 | Cleavage makes mature APOD smaller than the precursor |
| Disulfide-linked homodimer | Can yield a band near twice the monomer size under non-reducing conditions |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | APOD is secreted | Test conditioned medium or plasma alongside the lysate |
| Band higher than expected | Complex N-linked glycans or a homodimer may contribute | Compare reducing and non-reducing lanes and use an APOD identity control |
| Band lower than expected | Signal-peptide cleavage produces mature APOD | Compare with a suitable APOD control and verify band identity |
| Broad smear instead of sharp band | Variable glycosylation at Asn65 and Asn98 is possible | Compare untreated and deglycosylated samples with an APOD identity control |
| Multiple bands | Monomer and disulfide-linked complexes may coexist | Compare reducing and non-reducing lanes and confirm APOD identity |
| Weak or no signal | Secreted APOD may be scarce in the sampled cell fraction | Check conditioned medium or plasma with a positive control |
Experiment evidence
Western blot images showing APOD band
M02196 · APOD
PA1388 · APOD
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for APOD Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for APOD in Western blot, you can use adrenal gland tissue, where HPA reports high expression.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Cervix (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: APOD is secreted, so conditioned medium may give a stronger signal than whole-cell lysate.
HPA tissue expression evidence for APOD
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | glial cells | High | Protein (IHC) | HPA → |
| Cerebellum | granular cells - cytoplasm/membrane | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Cervix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Duodenum | endocrine cells | Not detected | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Not detected | Protein (IHC) | HPA → |
| Fallopian tube | glandular cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced APOD Western Blot Tips
Deeper troubleshooting and optimisation questions for APOD, answered from its protein features.
How should APOD 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.
Does APOD have alternative isoforms to consider?
Isoforms · The supplied record lists one isoform and no alternative sequence. Do not assign an unexpected band to another APOD isoform on the basis of these features.
Do annotated modifications prove a band shift?
PTM · The linked UniProt record describes protein features. A modification annotation alone does not demonstrate a visible shift; retain any condition or experimental qualifier attached to it.
Does this guide establish induction of APOD?
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 APOD?
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 M02196 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 APOD bands be quantified?
Quantitation · Because APOD can form a homodimer and a plasma heterodimer with APOA2, define which band or bands you quantify and use the same definition across samples. Keep sample fraction and reducing conditions consistent when comparing APOD signal.
Why might APOD migrate differently from its predicted 21.3 kDa mass?
Interpretation · The 21.3 kDa prediction does not establish where APOD will migrate. APOD has a signal peptide at residues 1–20 and complex N-linked glycosylation sites at Asn65 and Asn98. These features are relevant when interpreting apparent mass, but the supplied evidence gives no observed band size or demonstrated shift.
APOD is secreted and has a signal peptide at residues 1–20. Its modified residue at UniProt position 21 is pyrrolidone carboxylic acid, and it has complex N-linked glycosylation at Asn65 and Asn98. Use these UniProt coordinates when comparing annotations; antibody or paper numbering may differ.
APOD is reported as a homodimer and, in plasma, as a disulfide-linked heterodimer with APOA2. Consider these forms when interpreting a higher band, especially in plasma samples. A band’s position alone does not identify its components.
APOD has three annotated disulfide bonds, and its plasma heterodimer with APOA2 is disulfide-linked. Compare reducing and nonreducing conditions when investigating higher bands; the supplied features do not establish which form produced any particular band.
APOD is annotated as secreted. Consider the extracellular fraction or medium alongside cell material when choosing samples, and record which fraction was loaded before comparing band intensity.
APOD is secreted and has a signal peptide at residues 1–20. Its modified residue at UniProt position 21 is pyrrolidone carboxylic acid, and it has complex N-linked glycosylation at Asn65 and Asn98. Use these UniProt coordinates when comparing annotations; antibody or paper numbering may differ.
APOD is reported as a homodimer and, in plasma, as a disulfide-linked heterodimer with APOA2. Consider these forms when interpreting a higher band, especially in plasma samples. A band’s position alone does not identify its components.
APOD has three annotated disulfide bonds, and its plasma heterodimer with APOA2 is disulfide-linked. Compare reducing and nonreducing conditions when investigating higher bands; the supplied features do not establish which form produced any particular band.
APOD is annotated as secreted. Consider the extracellular fraction or medium alongside cell material when choosing samples, and record which fraction was loaded before comparing band intensity.
Boster reagents
APOD 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-ApoD Rabbit Monoclonal Antibody
Cat # M02196
Real WB data
Anti-Apolipoprotein D/APOD Antibody Picoband®
Cat # PA1388
Both listed the supplier anti-APOD antibodies report human reactivity and have Western blot images. M02196 is shown with human plasma lysate; PA1388 is shown with MCF-7, HELA, and SMMC cell lysates. These captions document specific tested samples.
Which to pick: For human plasma, consider M02196, whose WB image uses plasma lysate. For cell lysates, consider PA1388, whose WB image shows MCF-7, HELA, and SMMC lysates. Both have WB images; choose based on the sample context closest to yours.
Source: BosterBio APOD gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P05090.
- Human Protein Atlas. APOD tissue expression.
- PMC9167181 — target-verified WB comparison
- PMC7091089 — target-verified WB comparison
- PMC2784685 — target-verified WB comparison