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- Table of Contents
SSRP1 / FACT complex subunit SSRP1 · Western blot design guide
Design a Western Blot for SSRP1
Real validated SSRP1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SSRP1 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
SSRP1 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 | ~81.1 kDa | |
| Observed band | ~92 kDa | |
| Gel | 5–20% (catalog A02606-1) | |
| Positive control | Cerebellum (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 SSRP1 Western Blot Protocols
The A02606-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human DLD-1, human MOLT-4 (catalog A02606-1) |
| Gel % | 5–20% (catalog A02606-1) |
| Load | 30 ug; reducing conditions (catalog A02606-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A02606-1) |
| Membrane | nitrocellulose membrane (catalog A02606-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A02606-1) |
| Primary antibody | A02606-1 · 0.5 μg/mL (catalog A02606-1) |
| Primary incubation | overnight at 4°C (catalog A02606-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A02606-1) |
| Secondary incubation | 1.5 hour at RT (catalog A02606-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A02606-1) |
| Detection | ECL (catalog A02606-1) |
Section 2
What Is the Expected SSRP1 Western Blot Band Size?
SSRP1 is predicted at 81.1 kDa but observed near 92 kDa on reducing blots; the cause of the difference is not established.
| Band near 92 kDa | Empirical SSRP1 band in reducing whole-cell lysates; confirm identity with antibody controls |
| Band near 81 kDa | Near the 81.1 kDa sequence-predicted mass; identity requires validation |
| Weak band in whole-cell lysate | Nuclear and chromatin-associated SSRP1 may be poorly recovered |
| Additional bands near the main band | Modified SSRP1 is possible, but distinct migration has not been demonstrated |
💡Expected SSRP1 appearanceSSRP1 has a predicted mass of 81.1 kDa, while reducing whole-cell blots show a band near 92 kDa; confirm band identity with ordinary controls because the cause of the difference is unestablished.
| UniProt predicted mass | 81.1 kDa from the protein sequence; observed migration is about 92 kDa |
| Phosphothreonine at residue 170 | May alter migration, but no visible shift is established |
| N6-acetyllysine at residue 233 | May alter migration, but no visible shift is established |
| Phosphoserine at residue 437 | May alter migration, but no visible shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear or chromatin-associated SSRP1 may be poorly recovered | Check nuclear extraction and a positive-control lysate |
| Band higher than expected | The empirical SSRP1 band is near 92 kDa; its difference from 81.1 kDa is unexplained | Compare with a positive-control lysate and confirm identity by SSRP1 depletion |
| Band lower than expected | The band may be a fragment or a nonspecific signal; no cleavage product is specified | Check sample integrity and confirm identity by SSRP1 depletion |
| Multiple bands | Modification states are possible, but their migration is unestablished | Compare bands after SSRP1 depletion and assess phosphatase sensitivity if relevant |
| Weak or no signal | Nuclear or chromatin-associated protein may be underrepresented in the preparation | Check loading and recovery with a nuclear marker |
Experiment evidence
Western blot images showing SSRP1 band
A02606-1 · SSRP1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SSRP1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SSRP1 in Western blot, you can use cerebellum tissue, where HPA reports high expression.
Positive control: Cerebellum (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: SSRP1 is nuclear, and HPA identifies adipose tissue as a possible negative control, though its absence should be verified by Western blot.
HPA tissue expression evidence for SSRP1
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 |
|---|---|---|---|---|
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | High | Protein (IHC) | HPA → |
| Placenta | trophoblastic cells | High | Protein (IHC) | HPA → |
| Skin | epidermal cells | High | Protein (IHC) | HPA → |
| Stomach | 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 → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Kidney | cells in glomeruli | Not detected | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SSRP1 Western Blot Tips
Deeper troubleshooting and optimisation questions for SSRP1, answered from its protein features.
How should SSRP1 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 SSRP1 isoforms explain multiple bands?
Isoforms · The supplied UniProt record lists one isoform and no alternative sequence. Do not assign additional bands to isoforms from this evidence alone; verify their identity before quantifying them as SSRP1.
Which phosphorylation sites matter when comparing SSRP1 bands?
PTM · UniProt lists phosphothreonine 170; phosphotyrosines 441 and 452; and phosphoserines 437, 444, 471, 510, 657, 659, 667, 668, 671, 672, 673 and 688. These are UniProt coordinates, which may differ from antibody or paper numbering. The mapped sites support considering phosphorylation, but do not establish that it causes band separation.
UniProt assigns CK2 to phosphoserines 510 and 688 and lists SSRP1 in a CK2-SPT16-SSRP1 complex. These features motivate checking phosphorylation when comparing conditions involving CK2, but they do not establish a condition-dependent band shift. Keep the UniProt site numbering when comparing antibody specifications.
UniProt records N-acetylalanine at position 2 and N6-acetyllysine at positions 233, 413 and 542. Use those UniProt coordinates when assessing site-specific reagents. Acetylation at these sites does not, by itself, establish a resolvable Western-blot shift.
UniProt assigns CK2 to phosphoserines 510 and 688 and lists SSRP1 in a CK2-SPT16-SSRP1 complex. These features motivate checking phosphorylation when comparing conditions involving CK2, but they do not establish a condition-dependent band shift. Keep the UniProt site numbering when comparing antibody specifications.
UniProt records N-acetylalanine at position 2 and N6-acetyllysine at positions 233, 413 and 542. Use those UniProt coordinates when assessing site-specific reagents. Acetylation at these sites does not, by itself, establish a resolvable Western-blot shift.
Does this guide establish induction of SSRP1?
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 SSRP1?
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 A02606-1 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 SSRP1 be sampled for consistent quantitation?
Quantitation · SSRP1 is reported in the nucleus, nucleolus and on chromosomes, including chromatin at actively transcribed loci. Use the same fractionation and extraction procedure across samples, and compare bands from equivalent fractions with consistent loading. Differences in recovery could affect an apparent abundance comparison.
Why might SSRP1 appear near 92 kDa instead of 81.1 kDa?
Interpretation · The supplied apparent band is about 92 kDa, while the predicted mass is 81.1 kDa. SSRP1 has mapped modifications, but their presence alone does not explain the difference or establish a visible shift. Use the observed position as a reference and verify band identity independently.
The record provides no signal peptide, propeptide, glycosylation sites or alternative sequence to assign an extra band to those features. It does list phosphorylation and acetylation, but neither proves the identity of another band. Verify unexpected bands independently before including them in SSRP1 measurements.
The record provides no signal peptide, propeptide, glycosylation sites or alternative sequence to assign an extra band to those features. It does list phosphorylation and acetylation, but neither proves the identity of another band. Verify unexpected bands independently before including them in SSRP1 measurements.
Boster reagents
SSRP1 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-SSRP1 Antibody Picoband®
Cat # A02606-1
A02606-1 is a rabbit polyclonal anti-SSRP1 antibody with a Western blot image showing human, monkey, rat, and mouse specimens. The reported band is approximately 92 kDa, versus an expected 81 kDa. The supplied evidence is limited to this product image.
Which to pick: A02606-1 is the only listed option. It reports human, monkey, mouse, and rat reactivity and has a Western blot image. Check the caption for the tested specimens and conditions when planning your sample.
Source: BosterBio SSRP1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q08945.
- Human Protein Atlas. SSRP1 tissue expression.
- PMC5780019 — target-verified WB comparison
- PMC8215854 — target-verified WB comparison
- PMC6023160 — target-verified WB comparison