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- Table of Contents
PYGL / Glycogen phosphorylase, liver form · Western blot design guide
Design a Western Blot for PYGL
Real validated PYGL Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PYGL 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
PYGL 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 | ~97.1 kDa | |
| Observed band | ~97 kDa | |
| Gel | 10% (catalog A06317-1) | |
| Positive control | Bone marrow (IHC candidate; verify WB) +4 more | |
| Negative control | Bronchus (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated PYGL Western Blot Protocols
The A06317-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hacat, human PC-3, rat liver (catalog A06317-1) |
| Gel % | 10% (catalog A06317-1) |
| Load | 30 ug; reducing conditions (catalog A06317-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A06317-1) |
| Membrane | nitrocellulose membrane (catalog A06317-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A06317-1) |
| Primary antibody | A06317-1 · 0.5 μg/mL (catalog A06317-1) |
| Primary incubation | overnight at 4°C (catalog A06317-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A06317-1) |
| Secondary incubation | 1.5 hour at RT (catalog A06317-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A06317-1) |
| Detection | ECL (catalog A06317-1) |
Section 2
What Is the Expected PYGL Western Blot Band Size?
PYGL is predicted at 97.1 kDa and observed at ~97 kDa; these values agree within rounding, with no migration offset established.
| Band at ~97 kDa | Matches the empirical PYGL band and its 97.1 kDa predicted mass. |
| Band near twice the monomer size under nonreducing conditions | Could represent the annotated PYGL homodimer; confirm with reduction and band-identity controls. |
| Additional band at a different size | Could reflect isoform 1 or 2, but their masses and separation are not supplied. |
| Band near ~97 kDa in a cytosolic fraction | Consistent with PYGL's cytosolic location; confirm band identity. |
💡Expected PYGL appearancePYGL is predicted at 97.1 kDa and observed at ~97 kDa in reducing blots; confirm identity with appropriate antibody and sample controls.
| Predicted PYGL mass | Places the monomer near 97.1 kDa, consistent with the observed ~97 kDa band. |
| Homodimer formation | Could yield a band near twice the monomer size if the dimer persists during electrophoresis. |
| Splice isoform 1 | May differ in size from isoform 2; its individual mass and migration are not supplied. |
| Splice isoform 2 | May differ in size from isoform 1; its individual mass and migration are not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | PYGL is cytosolic; sample abundance or recovery may be insufficient. | Check a positive lysate and cytosolic extraction, then verify transfer and antibody performance. |
| Band higher than expected | A PYGL homodimer may persist if reduction is incomplete. | Compare reducing and nonreducing samples and confirm band identity. |
| Band lower than expected | The supplied features do not establish a smaller mature product; degradation or nonspecific binding is possible. | Use fresh lysate with protease inhibitors and an independent antibody or knockdown control. |
| Multiple bands | Two splice isoforms are annotated, but their migration difference is unknown. | Confirm each band with an independent antibody or knockdown control. |
| Weak or no signal | PYGL abundance or recovery in the tested sample may be low. | Check sample loading, transfer, and a positive PYGL lysate. |
| Fragments below expected size | Sample proteolysis is possible; no signal-peptide or propeptide cleavage is annotated. | Prepare fresh lysate with protease inhibitors and verify fragments with an independent antibody. |
Experiment evidence
Western blot images showing PYGL band
A06317-1 · PYGL
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PYGL Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PYGL in Western blot, you can use bone marrow lysate, which has high HPA expression.
Positive control: Bone marrow (IHC candidate; verify WB)
Negative control: Bronchus (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Cytosolic PYGL should be accessible in tissue lysate, and HPA lists bronchus as a negative tissue.
HPA tissue expression evidence for PYGL
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 |
|---|---|---|---|---|
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Liver | hepatocytes | High | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Lung | macrophages | Medium | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Cervix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Duodenum | glandular cells | Not detected | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PYGL Western Blot Tips
Deeper troubleshooting and optimisation questions for PYGL, answered from its protein features.
How should PYGL 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 PYGL isoforms produce different bands?
Isoforms · UniProt lists two isoforms. Isoform 2 lacks residues 82–115 relative to isoform 1, so it may migrate differently. Confirm isoform identity before assigning a second band; the sequence difference alone does not predict its exact gel position.
How should PYGL phosphorylation be assessed?
PTM · UniProt lists phosphoserine at positions 15, 524, 561, and 639. PHK phosphorylates Ser15 in phosphorylase a; PPP1R3B recruits PP1, which dephosphorylates and inactivates PYGL. For a phospho-specific readout, compare it with total PYGL. Positions use UniProt numbering, which may differ from antibody or paper numbering.
UniProt lists N-acetylalanine at position 2, succinyllysine at 364, acetyllysine at 470 and 796, and pyridoxal phosphate–modified lysine at 681. These features alone neither establish a visible band shift nor explain a difference between apparent and predicted mass.
UniProt lists N-acetylalanine at position 2, succinyllysine at 364, acetyllysine at 470 and 796, and pyridoxal phosphate–modified lysine at 681. These features alone neither establish a visible band shift nor explain a difference between apparent and predicted mass.
Does this guide establish induction of PYGL?
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 PYGL Western blot?
Transfer · PYGL is a 97.1 kDa cytosolic protein. Use transfer conditions suitable for a protein near 97 kDa and check that protein transfers to the membrane. The supplied features do not specify a membrane or transfer setting.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A06317-1 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 can PYGL abundance be quantified across samples?
Quantitation · Measure the PYGL band near 97 kDa within the assay’s linear range and normalize consistently across samples. If measuring phosphorylase a, assess Ser15 phosphorylation separately from total PYGL so a change in phosphorylation is not mistaken for a change in abundance. Ser15 uses UniProt numbering.
Why is the PYGL band near 97 kDa?
Interpretation · The observed band near 97 kDa agrees with the 97.1 kDa predicted mass. Listed modifications do not, by themselves, establish a visible mobility shift.
First compare them with the ~97 kDa band and consider isoform 2, which lacks residues 82–115. PYGL is an active homodimer, but that feature alone does not identify a higher band on a blot. The listed modifications likewise do not establish visible shifts. Confirm band identity before assigning an isoform, dimer, or modified species.
First compare them with the ~97 kDa band and consider isoform 2, which lacks residues 82–115. PYGL is an active homodimer, but that feature alone does not identify a higher band on a blot. The listed modifications likewise do not establish visible shifts. Confirm band identity before assigning an isoform, dimer, or modified species.
Boster reagents
PYGL 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-PYGL Antibody Picoband®
Cat # A06317-1
The catalog reports one anti-PYGL antibody, A06317-1, for Western blotting. Its image caption reports a band near the expected 97 kDa in the tested human cell and rat and mouse tissue lysates. No additional antibody or publication evidence was supplied.
Which to pick: A06317-1 is the only listed option. It is catalogued as reactive with human, mouse, and rat, and has a Western blot image using human Hacat and PC-3 cells, rat liver and lung, and mouse liver lysates.
Source: BosterBio PYGL gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P06737.
- Human Protein Atlas. PYGL tissue expression.
- PMC6193451 — target-verified WB comparison
- PMC11543057 — target-verified WB comparison
- PMC13007716 — target-verified WB comparison