This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
SRSF7 / Serine/arginine-rich splicing factor 7 · Western blot design guide
Design a Western Blot for SRSF7
Source-linked SRSF7 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SRSF7 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
SRSF7 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 | ~27.4 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Lung (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 | Peptide-blocking control | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 4 isoform(s) |
Section 1
Source-Linked SRSF7 Western Blot Protocol Options
The A30768 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | HuvEc cells (catalog A30768) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A30768; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected SRSF7 Western Blot Band Size?
SRSF7 is predicted at 27.4 kDa; isoforms and modifications could affect migration, but no empirical band size or visible shift is established.
| Band near 27.4 kDa | Consistent with the predicted SRSF7 mass; identity needs confirmation |
| Several bands at different positions | Could reflect the four named isoforms; their migration is unestablished |
| Nearby doublet | Could reflect differing phosphorylation states; a visible shift is unproven |
| Signal in nuclear and cytoplasmic fractions | Consistent with both documented locations of SRSF7 |
💡Expected SRSF7 appearanceA band near the predicted 27.4 kDa is a starting point, not an empirical band assignment; confirm its identity with knockdown or peptide blocking.
| Predicted SRSF7 mass | 27.4 kDa is the sequence-based reference, not a measured band position |
| Isoform 1 | May differ in size from other isoforms; its relative mass is unspecified |
| Isoform 2 | May differ in size from other isoforms; its relative mass is unspecified |
| Isoform 3 | May differ in size from other isoforms; its relative mass is unspecified |
| Isoform 4 | May differ in size from other isoforms; its relative mass is unspecified |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | SRSF7 may be missed if nuclear protein is poorly extracted | Check nuclear extraction and a nuclear protein control |
| Band higher than expected | Documented phosphorylation could affect migration, but a shift is unproven | Compare phosphatase-treated and untreated samples and confirm band identity |
| Band lower than expected | An isoform may differ in size; isoform masses are unavailable | Confirm identity by SRSF7 knockdown and check the antibody epitope |
| Multiple bands | Four isoforms or differing phosphorylation states are possible | Use knockdown to identify SRSF7 bands; compare phosphatase-treated samples |
| Weak or no signal | Recovery may vary between nuclear and cytoplasmic fractions | Check both fractions and verify extraction and loading |
Experiment evidence
Western blot images showing SRSF7 band
A30768 · SRSF7
A30768 · SRSF7
A30768 · SRSF7
A30768 · SRSF7
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SRSF7 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SRSF7 in Western blot, you can use lung tissue, which HPA rates High.
Positive control: Lung (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: SRSF7 is intracellular, so tissue lysate is suitable, and HPA lists a not-detected tissue for a negative control.
HPA tissue expression evidence for SRSF7
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 |
|---|---|---|---|---|
| Lung | alveolar cells type I | High | Protein (IHC) | HPA → |
| Skin | cells in granular layer | High | Protein (IHC) | HPA → |
| Testis | round or early spermatids | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | glandular 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 → |
| Spleen | cells in red pulp | Not detected | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | Low | Protein (IHC) | HPA → |
| Prostate | glandular cells | Low | Protein (IHC) | HPA → |
| Salivary gland | glandular cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced SRSF7 Western Blot Tips
Deeper troubleshooting and optimisation questions for SRSF7, answered from its protein features.
How should SRSF7 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.
How could SRSF7 isoforms affect band interpretation?
Isoforms · Four isoforms are listed. Relative to the canonical sequence, isoform 2 replaces residues 130–135 and lacks 136–238; isoform 3 replaces 130–132 and lacks 133–238; isoform 4 lacks 209–220. Consider these sequence differences when evaluating extra bands, and check whether the antibody recognizes a region retained in each isoform.
Possibly. Isoforms 2 and 3 lack canonical residues 136–238 and 133–238, respectively; isoform 4 lacks 209–220. Compare the band with the antibody's recognized region and the expected sequence changes. Band size alone cannot establish its identity.
Possibly. Isoforms 2 and 3 lack canonical residues 136–238 and 133–238, respectively; isoform 4 lacks 209–220. Compare the band with the antibody's recognized region and the expected sequence changes. Band size alone cannot establish its identity.
Which SRSF7 modifications matter when assessing band shifts?
PTM · UniProt lists acetyllysine at position 24 and phosphoserines at 32, 163, 165, 167, 181, 183, 192, 194, 196, 231, and 233. These are canonical UniProt coordinates; numbering in papers or antibody materials may differ. Their presence alone does not establish that a shifted band will be visible.
Does this guide establish induction of SRSF7?
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 SRSF7 Western blot?
Transfer · The canonical protein has a predicted mass of 27.4 kDa, while isoforms 2 and 3 lack substantial C-terminal sequence. Choose transfer conditions that retain proteins around 27.4 kDa and smaller, then check transfer with a suitable size marker. The supplied features do not identify a uniquely preferred transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A30768 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 SRSF7 bands be quantified?
Quantitation · Define which bands the antibody detects before combining their signals: four isoforms are listed, and their sequences differ. SRSF7 is reported in both nucleus and cytoplasm, so compare like fractions across samples. The supplied features give no observed band position with which to assign every band.
Should SRSF7 migrate at exactly 27.4 kDa?
Interpretation · 27.4 kDa is the predicted mass, and no observed band position is supplied. Use it as a reference, not an exact migration target. The listed modifications and isoforms do not, by themselves, establish a visible shift or explain any difference from 27.4 kDa.
SRSF7 is reported in large complexes containing CCNL1 and p110 isoforms of CDC2L1 or CDC2L2, and it interacts with CCNL2 and CPSF6. Those interactions do not establish that a high molecular weight band is SRSF7 or that a complex persists under the blot's preparation conditions. Verify the band independently.
SRSF7 is reported in large complexes containing CCNL1 and p110 isoforms of CDC2L1 or CDC2L2, and it interacts with CCNL2 and CPSF6. Those interactions do not establish that a high molecular weight band is SRSF7 or that a complex persists under the blot's preparation conditions. Verify the band independently.
Boster reagents
SRSF7 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-SFRS7 Antibody
Cat # A30768
The catalog reports A30768, an anti-SRSF7 antibody with Human and Mouse reactivity. Its Western blot image is described as an analysis of HuvEc cells; the supplied evidence does not show a Mouse blot.
Which to pick: A30768 is the only listed option and has a Western blot image from HuvEc cells. For Mouse samples, reactivity is listed, but no Mouse blot is supplied.
Source: BosterBio SRSF7 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q16629.
- Human Protein Atlas. SRSF7 tissue expression.
- PMC5844074 — target-verified WB comparison
- PMC11056844 — target-verified WB comparison
- PMC10787126 — target-verified WB comparison