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- Table of Contents
AGPS / Alkyldihydroxyacetonephosphate synthase, peroxisomal · Western blot design guide
Design a Western Blot for AGPS
Source-linked AGPS Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-AGPS 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
AGPS 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 | ~72.9 kDa | |
| Observed band | ~73 kDa | |
| Gel | 5–20% (catalog A03481-1) | |
| Positive control | Bronchus (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 | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked AGPS Western Blot Protocol Options
The A03481-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human K562, human HepG2, human RT4, human HEL, human CACO-2, human SiHa (catalog A03481-1) |
| Gel % | 5–20% (catalog A03481-1) |
| Load | 30 ug; reducing conditions (catalog A03481-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A03481-1) |
| Membrane | nitrocellulose membrane (catalog A03481-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A03481-1) |
| Primary antibody | A03481-1 · 0.25 μg/mL (catalog A03481-1) |
| Primary incubation | overnight at 4°C (catalog A03481-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A03481-1) |
| Secondary incubation | 1.5 hour at RT (catalog A03481-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A03481-1) |
| Detection | ECL (catalog A03481-1) |
Section 2
What Is the Expected AGPS Western Blot Band Size?
AGPS is predicted at 72.9 kDa and observed at ~73 kDa; the supplied evidence does not establish a cause for the small difference.
| Band at ~73 kDa | Matches the empirical AGPS band in reducing whole-cell lysates and its 72.9 kDa predicted monomer mass. |
| Band near ~146 kDa | Could reflect AGPS homodimers if they remain associated during electrophoresis; confirm band identity. |
| Bands near ~73 and ~146 kDa | Could reflect monomers alongside retained AGPS homodimers; confirm both bands. |
| Weak band in a peroxisome-depleted fraction | Consistent with AGPS localization to peroxisomes and their membranes. |
💡Expected AGPS appearanceAGPS has a predicted monomer mass of 72.9 kDa and an empirical band at ~73 kDa in reducing whole-cell lysates; confirm band identity with antibody and sample controls.
| Predicted AGPS monomer mass | 72.9 kDa, consistent with the empirical ~73 kDa band. |
| Homodimer formation | May produce a band near twice the monomer mass if the dimer survives sample preparation. |
| Phosphoserine 65 and phosphothreonine 74 | These modifications are annotated; a visible migration change is not established. |
| N6-acetyllysine 102 and 347 | These modifications are annotated; a visible migration change is not established. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | AGPS may be underrepresented after extraction of peroxisomes or their membranes. | Check fraction recovery and compare with a peroxisome-enriched preparation. |
| Band higher than expected | The annotated homodimer may remain associated during sample preparation. | Compare reducing and nonreducing preparations and verify band identity. |
| Multiple bands | Monomer and retained homodimer are possible; other bands are not explained by the supplied features. | Compare sample preparation conditions and use an AGPS depletion control. |
| Weak or no signal | Peroxisomal AGPS may be poorly recovered during extraction. | Check extraction and loading with peroxisomal and total-protein controls. |
| Band lower than expected | No supplied feature establishes a smaller AGPS band. | Check band identity with AGPS depletion and an independent antibody. |
Experiment evidence
Western blot images showing AGPS band
A03481-1 · AGPS
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for AGPS Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for AGPS in Western blot, you can use bronchus tissue, which shows high HPA expression.
Positive control: Bronchus (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: HPA identifies both high-expression and not-detected tissues, making tissue controls feasible for peroxisomal AGPS.
HPA tissue expression evidence for AGPS
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 |
|---|---|---|---|---|
| Bronchus | ciliated cells (ciliary rootlets) | High | Protein (IHC) | HPA → |
| Fallopian tube | ciliated cells (ciliary rootlets) | High | Protein (IHC) | HPA → |
| Testis | sertoli cells | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Purkinje 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 → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced AGPS Western Blot Tips
Deeper troubleshooting and optimisation questions for AGPS, answered from its protein features.
How should AGPS 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 AGPS isoforms explain additional bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It provides no isoform-based explanation for additional bands; check their specificity experimentally.
Which AGPS modifications are listed, and do they shift the band?
PTM · The listed UniProt coordinates are phosphoserine 65, phosphothreonine 74, and N6-acetyllysine 102 and 347. These features alone do not show that any modification produces a visible band shift. Keep the UniProt numbering explicit when comparing antibody or paper coordinates.
Does this guide establish induction of AGPS?
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 AGPS Western blot?
Transfer · The record gives a predicted mass of 72.9 kDa but no transfer method. Check transfer of the ~73 kDa region with a membrane stain and adjust the method empirically if signal is weak.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A03481-1 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 I quantify the AGPS band?
Quantitation · Quantify the ~73 kDa band within the detection system’s linear range and normalize for lane loading. AGPS is annotated in the peroxisome and peroxisome membrane, so compare samples prepared with the same fractionation procedure.
Does the ~73 kDa AGPS band match its predicted mass?
Interpretation · Yes. The observed ~73 kDa band is close to AGPS’s predicted 72.9 kDa mass. The listed features do not establish a meaningful mass shift.
AGPS is annotated as a homodimer by similarity to another UniProt entry. That annotation does not establish that a higher band contains AGPS or that the dimer survives blot preparation. Verify the band’s identity experimentally.
AGPS is annotated in the peroxisome and peroxisome membrane by similarity. Comparing matched whole-cell and peroxisome-enriched samples may help assess whether an unexpected band tracks with AGPS, but localization alone cannot identify it.
AGPS is annotated as a homodimer by similarity to another UniProt entry. That annotation does not establish that a higher band contains AGPS or that the dimer survives blot preparation. Verify the band’s identity experimentally.
AGPS is annotated in the peroxisome and peroxisome membrane by similarity. Comparing matched whole-cell and peroxisome-enriched samples may help assess whether an unexpected band tracks with AGPS, but localization alone cannot identify it.
Boster reagents
AGPS 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-AGPS Antibody Picoband®
Cat # A03481-1
The catalog reports one anti-AGPS antibody, A03481-1, with human reactivity. Its Western blot image shows an approximately 73 kDa band in seven human cell lysates under the reported conditions. No publication or independent validation evidence is supplied.
Which to pick: A03481-1 is the only listed option. It has a Western blot image from HeLa, K562, HepG2, RT4, HEL, CACO-2, and SiHa whole cell lysates; use the reported sample and conditions to judge its fit for your experiment.
Source: BosterBio AGPS gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O00116.
- Human Protein Atlas. AGPS tissue expression.
- PMC10782585 — target-verified WB comparison
- PMC10003153 — target-verified WB comparison
- PMC10907675 — target-verified WB comparison