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- Table of Contents
PGAM1 / Phosphoglycerate mutase 1 · Western blot design guide
Design a Western Blot for PGAM1
Real validated PGAM1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PGAM1 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
PGAM1 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 | ~28.8 kDa | |
| Observed band | ~29 kDa | |
| Gel | 10% (catalog A04470-2) | |
| Positive control | Kidney (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 Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated PGAM1 Western Blot Protocols
The A04470-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human 293T, human Hela, human HepG2 (catalog A04470-2) |
| Gel % | 10% (catalog A04470-2) |
| Load | 30 ug; reducing conditions (catalog A04470-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A04470-2) |
| Membrane | nitrocellulose membrane (catalog A04470-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A04470-2) |
| Primary antibody | A04470-2 · 0.25 μg/mL (catalog A04470-2) |
| Primary incubation | overnight at 4°C (catalog A04470-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A04470-2) |
| Secondary incubation | 1.5 hour at RT (catalog A04470-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A04470-2) |
| Detection | ECL (catalog A04470-2) |
Section 2
What Is the Expected PGAM1 Western Blot Band Size?
PGAM1 is predicted at 28.8 kDa and observed at ~29 kDa; these values agree within rounding, and no distinct migration cause is established.
| Dominant band at ~29 kDa | PGAM1 monomer, consistent with the 28.8 kDa predicted mass and the observed band |
| Higher band near ~58 kDa | Could reflect the UniProt homodimer if it survives sample preparation; confirm its identity |
| Close doublet near ~29 kDa | Could reflect different modification states, but their separation is not established |
| Single ~29 kDa band without a higher band | Consistent with dissociation of the homodimer during denaturing electrophoresis |
💡Expected PGAM1 appearancePGAM1 is predicted at 28.8 kDa and was observed at ~29 kDa in reducing whole-cell lysates; use an appropriate band-identity control to verify a signal in new samples.
| Predicted monomer mass | Places the expected monomer band near 28.8 kDa |
| Homodimer formation | Could yield a band near twice the monomer mass if the complex survives electrophoresis |
| Homodimer dissociation | Favors migration at the monomer size during denaturing electrophoresis |
| Monomer mass in reducing lysates | Is consistent with the observed ~29 kDa band |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | A homodimer may persist if denaturation is incomplete | Repeat with fresh denaturing sample buffer and verify band identity with a PGAM1-specific control |
| Multiple bands | A persistent homodimer or different modification states are possible | Compare denaturing conditions and verify each band with a PGAM1-specific control |
| Band lower than expected | A smaller band is not explained by the supplied PGAM1 processing features | Check sample integrity and verify the band with a PGAM1-specific control |
| Weak or no signal | Target amount or assay sensitivity may be insufficient | Include a positive lysate and check loading, transfer, and antibody detection |
| Fragments below expected size | Sample degradation is possible; no cleavage product is specified | Prepare fresh lysate with protease inhibitors and confirm fragment identity |
Experiment evidence
Western blot images showing PGAM1 band
A04470-2 · PGAM1
A04470-2 · PGAM1
M04470-1 · PGAM1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PGAM1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PGAM1 in Western blot, you can use kidney tissue.
Positive control: Kidney (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: Kidney is high and adipose tissue is not detected, making tissue controls feasible.
HPA tissue expression evidence for PGAM1
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 |
|---|---|---|---|---|
| Kidney | cells in tubules | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | neuropil | 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 → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Duodenum | glandular cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PGAM1 Western Blot Tips
Deeper troubleshooting and optimisation questions for PGAM1, answered from its protein features.
Where should the main PGAM1 band appear?
Band shift · PGAM1 has a predicted mass of 28.8 kDa, consistent with the observed band near 29 kDa. Use that region to identify the main band; a small difference between predicted and apparent mass does not establish a modification.
Could another PGAM1 isoform explain a second band?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no documented PGAM1 isoform to assign to a second band.
Could PGAM1 phosphorylation cause a band shift?
PTM · UniProt lists phosphoserine at positions 14, 23, 31, and 118, and phosphotyrosine at 26. These are UniProt coordinates; antibody or paper numbering may differ. The listed sites alone do not establish a visible shift.
UniProt lists N6-acetyllysine at positions 106, 251, 253, and 254, with N6-succinyllysine as an alternate modification at 251. These are UniProt coordinates. Their presence does not by itself explain an unexpected band or establish a detectable mass shift.
The supplied record lists no glycosylation sites, signal peptide, or propeptide. It provides no basis to attribute a higher band to glycosylation or precursor processing.
UniProt lists N6-acetyllysine at positions 106, 251, 253, and 254, with N6-succinyllysine as an alternate modification at 251. These are UniProt coordinates. Their presence does not by itself explain an unexpected band or establish a detectable mass shift.
The supplied record lists no glycosylation sites, signal peptide, or propeptide. It provides no basis to attribute a higher band to glycosylation or precursor processing.
Does this guide establish induction of PGAM1?
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 PGAM1?
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 A04470-2 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.
Which band should be quantified for PGAM1?
Quantitation · Quantify the consistently identified main band near 29 kDa, matching the predicted 28.8 kDa mass. Treat additional bands separately unless their identity is established; the listed modifications and homodimer do not establish what those bands represent.
How should unexpected high-mass PGAM1 bands be interpreted?
Interpretation · PGAM1 is reported as a homodimer, but that feature alone cannot identify a high-mass Western blot band as a dimer. Keep the assignment tentative and assess it against the main band near 29 kDa.
Boster reagents
PGAM1 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-PGAM1 Antibody Picoband®
Cat # A04470-2
Real WB data
Anti-PGAM1 Rabbit Monoclonal Antibody
Cat # M04470-1
Two the supplier anti-PGAM1 antibodies are listed for Western blotting, both with human, mouse, and rat reactivity and WB images. A04470-2 shows a band near the expected 29 kDa in the captioned lysates; M04470-1 shows A431 cell lysate. No independent validation is supplied.
Which to pick: Choose A04470-2 if you want documented WB conditions and examples from human cell lines plus rat and mouse brain and liver. M04470-1 has a WB image for A431 cell lysate, but its supplied caption gives fewer experimental details.
Source: BosterBio PGAM1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P18669.
- Human Protein Atlas. PGAM1 tissue expression.
- PMC5605442 — target-verified WB comparison
- PMC8242254 — target-verified WB comparison
- PMC11151566 — target-verified WB comparison