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- Table of Contents
SALL4 / Sal-like protein 4 · Western blot design guide
Design a Western Blot for SALL4
Real validated SALL4 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SALL4 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
SALL4 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 | ~112.2 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Ovary (IHC candidate; verify WB) +1 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated SALL4 Western Blot Protocols
The A01827 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of various cell lines, (catalog A01827) |
| Gel % | 8–10% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Wet/tank transfer; optimize duration (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A01827; 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 SALL4 Western Blot Band Size?
SALL4 has a predicted mass of 112.2 kDa; its isoforms and phosphorylation could affect migration, but no empirical band or feature-driven shift is established.
| Band near 112.2 kDa | Consistent with the predicted SALL4 mass; confirm identity with a control |
| Additional band at a different mass | Could reflect SALL4A or SALL4B; their migration is not established |
| Mobility-shifted band or doublet | Could reflect phosphorylation; a visible shift is not established |
| Weak band in a cytoplasmic fraction | SALL4 may be enriched in the nucleus |
💡Expected SALL4 appearanceUniProt predicts 112.2 kDa for SALL4, but no empirical band size is supplied; use knockdown or another band-identity control to assess any band near that size.
| Predicted SALL4 mass | 112.2 kDa is the sequence-based reference, not a measured band |
| SALL4A and SALL4B isoforms | Alternative splicing may alter size; their individual masses and separation are not supplied |
| Phosphoserines at residues 57, 307, 776, 789, 852 and 1019 | May affect migration, but no visible shift is established |
| Phosphothreonine at residue 541 | May affect migration, but no visible shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear SALL4 may be poorly recovered | Check nuclear extraction and use a positive control |
| Band higher than expected | Phosphorylation may affect migration, but the cause is unproven | Compare phosphatase-treated material and verify identity by knockdown |
| Band lower than expected | An alternative splice isoform is possible; its mass is unknown | Verify identity by knockdown or an isoform-specific reagent |
| Multiple bands | SALL4A and SALL4B are annotated, but distinct bands are unconfirmed | Use knockdown or isoform-specific controls to identify the bands |
| Weak or no signal | The preparation may contain little nuclear SALL4 | Check nuclear recovery and include a positive control |
Experiment evidence
Western blot images showing SALL4 band
A01827 · SALL4
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SALL4 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SALL4 in Western blot, you can use ovary tissue, which HPA rates High.
Positive control: Ovary (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: An HPA not-detected tissue such as adipose tissue provides a feasible negative control.
HPA tissue expression evidence for SALL4
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 |
|---|---|---|---|---|
| Ovary | follicle cells | High | Protein (IHC) | HPA → |
| Testis | spermatogonia 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 → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SALL4 Western Blot Tips
Deeper troubleshooting and optimisation questions for SALL4, answered from its protein features.
How should SALL4 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.
Can bands identify SALL4 isoforms?
Isoforms · Use the linked UniProt isoform annotations; migration alone does not identify an isoform. No additional isoform-specific band assignment is established here.
Which phosphorylation sites matter when interpreting SALL4 bands?
PTM · UniProt lists phosphoserine at positions 57, 307, 776, 789, 852 and 1019, plus phosphothreonine at 541. These are canonical UniProt coordinates; paper or antibody numbering may differ. Phosphorylation at an annotated site does not by itself establish a visible mobility shift.
SALL4B lacks canonical residues 385–821, which include phosphothreonine 541 and phosphoserines 776 and 789. The other listed sites lie outside that interval. Use canonical UniProt coordinates when comparing site claims across isoforms.
SALL4B lacks canonical residues 385–821, which include phosphothreonine 541 and phosphoserines 776 and 789. The other listed sites lie outside that interval. Use canonical UniProt coordinates when comparing site claims across isoforms.
Does this guide establish induction of SALL4?
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 SALL4?
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 A01827 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.
What should I consider when quantifying SALL4 bands?
Quantitation · Decide whether the measurement represents total detected SALL4 or one isoform. SALL4A and SALL4B differ by canonical residues 385–821, so antibody epitope coverage can affect which forms contribute to the signal. Apply the same band assignment across samples.
How should I interpret a band away from 112.2 kDa?
Interpretation · The supplied predicted mass is 112.2 kDa, but no observed band size is available. SALL4 has two isoforms and seven annotated phosphorylation sites. These features warrant checking isoform identity and phosphorylation, but neither proves a visible shift or explains a specific mass difference.
Yes. SALL4B lacks residues 385–821 of the canonical sequence, so it is expected to be smaller than SALL4A. Compare antibody epitope coverage before assigning a band: an epitope within that interval would not detect SALL4B.
Check whether the antibody epitope lies outside canonical residues 385–821, which SALL4B lacks. An antibody against the missing interval cannot recognize SALL4B through that epitope. Band size alone cannot establish isoform identity, especially without an observed reference size.
UniProt lists SALL4 in both the nucleus and cytoplasm. A signal in either fraction is therefore consistent with its annotated locations. Record which fraction was loaded when comparing band intensity across samples.
Yes. SALL4B lacks residues 385–821 of the canonical sequence, so it is expected to be smaller than SALL4A. Compare antibody epitope coverage before assigning a band: an epitope within that interval would not detect SALL4B.
Check whether the antibody epitope lies outside canonical residues 385–821, which SALL4B lacks. An antibody against the missing interval cannot recognize SALL4B through that epitope. Band size alone cannot establish isoform identity, especially without an observed reference size.
UniProt lists SALL4 in both the nucleus and cytoplasm. A signal in either fraction is therefore consistent with its annotated locations. Record which fraction was loaded when comparing band intensity across samples.
Boster reagents
SALL4 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-Sal-like protein 4 SALL4 Antibody
Cat # A01827
The catalog reports one anti-SALL4 antibody, A01827, with stated Human and Mouse reactivity. Its Western blot image is captioned as extracts from various cell lines; the supplied caption does not identify them or establish performance across all samples.
Which to pick: A01827 is the only listed option. It has a Western blot image and stated Human and Mouse reactivity; assess suitability for your sample and conditions from the full product details.
Source: BosterBio SALL4 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9UJQ4.
- Human Protein Atlas. SALL4 tissue expression.
- PMC7247129 — target-verified WB comparison
- PMC8639708 — target-verified WB comparison
- PMC6188178 — target-verified WB comparison