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- Table of Contents
PSPH / Phosphoserine phosphatase · Western blot design guide
Design a Western Blot for PSPH
Real validated PSPH Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PSPH 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
PSPH 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 | ~25 kDa | |
| Observed band | ~25 kDa | |
| Gel | 12% (catalog A05579-2) | |
| Positive control | Colon (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 | Acetylated | |
| Caveat | — | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated PSPH Western Blot Protocols
The A05579-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human A375, human HL-60, human MCF-7 (catalog A05579-2) |
| Gel % | 12% (catalog A05579-2) |
| Load | 30 ug; reducing conditions (catalog A05579-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A05579-2) |
| Membrane | nitrocellulose membrane (catalog A05579-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A05579-2) |
| Primary antibody | A05579-2 · 1:1000 (catalog A05579-2) |
| Primary incubation | overnight at 4°C (catalog A05579-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:1000 (catalog A05579-2) |
| Secondary incubation | 1.5 hour at RT (catalog A05579-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A05579-2) |
| Detection | ECL (catalog A05579-2) |
Section 2
What Is the Expected PSPH Western Blot Band Size?
PSPH is predicted at 25 kDa and observed at ~25 kDa; its homodimer could produce a higher band if retained during sample preparation.
| Single band at ~25 kDa | matches the predicted mass and the empirical band in reducing whole-cell lysates |
| Band near ~50 kDa | could reflect retained PSPH homodimer; its identity requires confirmation |
| Band at ~25 kDa in whole-cell lysate | is consistent with PSPH's cytosolic location |
| Little or no band in conditioned medium | is consistent with PSPH's cytosolic location |
💡Expected PSPH appearancePSPH is predicted at 25 kDa and observed at ~25 kDa in reducing whole-cell lysates; confirm band identity with an appropriate positive control and antibody specificity controls.
| UniProt predicted mass | places the monomer near 25 kDa, matching the observed ~25 kDa band |
| Homodimer formation | could yield a band near twice the monomer size if the dimer survives sample preparation |
| N-acetylmethionine at residue 1 | has no demonstrated resolvable effect on band size |
| Cytosolic location | supports detection in cell lysate but does not change apparent band size |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | PSPH signal may be below detection despite its cytosolic location | Check lysate loading and run a positive lysate control |
| Band higher than expected | PSPH homodimer may have survived sample preparation | Compare fully denatured reducing and nonreducing samples and confirm band identity |
| Band lower than expected | The band may be a degradation product or unrelated signal | Use fresh lysate and an antibody specificity control |
| Multiple bands | A retained homodimer or unrelated antibody signal may accompany the monomer | Compare sample preparation conditions and confirm the ~25 kDa band with a specificity control |
| Weak or no signal | PSPH abundance or sample loading may be insufficient | Check loading and include a positive lysate control |
| Fragments below expected size | PSPH may have degraded during lysate preparation | Prepare fresh lysate with protease inhibitors and compare the ~25 kDa signal |
Experiment evidence
Western blot images showing PSPH band
A05579-2 · PSPH
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PSPH Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PSPH in Western blot, you can use colon tissue, which HPA scores High.
Positive control: Colon (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.
⚠️Feasibility: Cytosolic PSPH is suited to lysate analysis, and HPA lists not-detected tissues as negative candidates.
HPA tissue expression evidence for PSPH
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 |
|---|---|---|---|---|
| Colon | glandular cells | High | Protein (IHC) | HPA → |
| Fallopian tube | ciliated cells (cell body) | High | Protein (IHC) | HPA → |
| Gallbladder | glandular cells | High | Protein (IHC) | HPA → |
| Kidney | cells in tubules | High | Protein (IHC) | HPA → |
| Prostate | glandular cells | High | 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 → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PSPH Western Blot Tips
Deeper troubleshooting and optimisation questions for PSPH, answered from its protein features.
How should PSPH 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 PSPH isoforms explain multiple bands?
Isoforms · The supplied UniProt record lists one isoform and no alternative sequence. These features do not support assigning additional bands to PSPH isoforms.
Does acetylation explain a shifted PSPH band?
PTM · UniProt lists N-acetylmethionine at position 1, using UniProt coordinates. That modification alone does not establish a visible band shift. Check the numbering convention before comparing position 1 with an antibody or paper annotation.
Does this guide establish induction of PSPH?
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 PSPH Western blot?
Transfer · PSPH is predicted at 25 kDa, so assess transfer efficiency around 25 kDa. The supplied features do not specify a membrane, transfer method, or settings; verify recovery of a band in that region under the chosen conditions.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A05579-2 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 as PSPH?
Quantitation · Start with the band near 25 kDa, matching both the predicted mass and supplied observed band. Verify the identity of any additional bands before including them in PSPH quantitation.
Should PSPH run at its predicted molecular mass?
Interpretation · PSPH is predicted at 25 kDa, and the supplied observed band is about 25 kDa. Use that region as the primary reference. Its listed features do not establish a visible shift from the predicted mass.
UniProt describes PSPH as a homodimer. A higher band could prompt investigation of its oligomeric state, but this feature alone does not identify that band. Confirm its identity before assigning it to PSPH.
UniProt places PSPH in the cytoplasm, specifically the cytosol. Check a cytosolic sample when assessing a weak or missing band near 25 kDa.
UniProt describes PSPH as a homodimer. A higher band could prompt investigation of its oligomeric state, but this feature alone does not identify that band. Confirm its identity before assigning it to PSPH.
UniProt places PSPH in the cytoplasm, specifically the cytosol. Check a cytosolic sample when assessing a weak or missing band near 25 kDa.
Boster reagents
PSPH 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-PSPH Antibody Picoband®
Cat # A05579-2
The catalog reports one anti-PSPH antibody, A05579-2. Its Western blot image reports a band near the expected 25 kDa in specified human, rat, and mouse lysates. No publication or independent validation evidence is supplied.
Which to pick: A05579-2 is the only listed choice. It is reported reactive with human, mouse, and rat; its WB image includes human cell lines, rat liver and cells, and mouse liver and HEPA1-6 lysates. Compare those tested samples with yours.
Source: BosterBio PSPH gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P78330.
- Human Protein Atlas. PSPH tissue expression.
- PMC6329917 — target-verified WB comparison
- PMC6715339 — target-verified WB comparison
- PMC3152677 — target-verified WB comparison