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SLC35D3 / Solute carrier family 35 member D3 · Western blot design guide
Design a Western Blot for SLC35D3
Source-linked SLC35D3 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SLC35D3 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled October 2026 · For research use; verify linked source records and product datasheet before use
SLC35D3 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 | ~44.2 kDa | |
| Observed band | 68 kDa | |
| Gel | 12–15% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | Band identity controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked SLC35D3 Western Blot Protocol Options
The A15258 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | HeLa cell lysate (catalog A15258) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A15258 · 1 μg/mL (catalog A15258) |
| 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 SLC35D3 Western Blot Band Size?
SLC35D3 has a predicted mass of 44.2 kDa and an empirical band at 68 kDa; the cause of their difference is not established.
| Band near 68 kDa | Empirical SLC35D3 band in cell lysate; confirm identity with antibody controls |
| Band near 44.2 kDa | Near the predicted sequence mass; identity requires validation |
| Weak band in whole-cell lysate | Membrane-associated SLC35D3 may be poorly recovered |
| Little signal in a soluble fraction | SLC35D3 is associated with vesicle, endosome, and endoplasmic reticulum membranes |
💡Expected SLC35D3 appearanceThe predicted mass is 44.2 kDa, while the empirical band is 68 kDa in cell lysate; the cause of the difference is unestablished, so confirm band identity with antibody controls.
| UniProt predicted molecular mass | Sets a sequence-based reference of 44.2 kDa |
| Calculated mass in daltons | The supplied 44,183 Da value corresponds to about 44.2 kDa |
| Full-length sequence mass | Does not establish where the protein migrates on a blot |
| Predicted mass versus empirical migration | The 44.2 kDa prediction does not explain the observed 68 kDa band |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Incomplete recovery of this multi-pass membrane protein | Check membrane solubilization and test a membrane-enriched fraction |
| Band higher than expected | The reported 68 kDa band exceeds the 44.2 kDa prediction for an unestablished reason | Confirm identity with an independent antibody or SLC35D3 depletion |
| Band lower than expected | Band identity or protein integrity is uncertain | Compare with a positive-control lysate and assess sample degradation |
| Multiple bands | Additional bands are not explained by the supplied single-isoform record | Check which bands respond to SLC35D3 depletion |
| Weak or no signal | Poor extraction of membrane-associated SLC35D3 may reduce recovery | Optimize membrane-protein extraction and check a positive-control lysate |
Experiment evidence
Western blot images showing SLC35D3 band
A15258 · SLC35D3
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SLC35D3 Western blot
🧪For positive controls for SLC35D3 in Western blot, you can use no HPA-supported tissue or cell line because no positive candidate was supplied.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA provides no tissue controls, and this multi-pass membrane protein may require membrane enrichment for detection.
HPA tissue expression evidence for SLC35D3
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced SLC35D3 Western Blot Tips
Deeper troubleshooting and optimisation questions for SLC35D3, answered from its protein features.
How should SLC35D3 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 SLC35D3 isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no isoform-based explanation for multiple bands. Check band specificity rather than assigning extra bands to isoforms.
Do listed modifications explain the SLC35D3 band position?
PTM · The supplied record lists no modified residues or glycosylation sites. Do not attribute the 68 kDa band to a specific modification on this evidence; the band position alone cannot establish one.
Does this guide establish induction of SLC35D3?
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 SLC35D3 Western blot?
Transfer · SLC35D3 is a multi-pass membrane protein with a predicted mass of 44.2 kDa and a supplied apparent band at 68 kDa. Optimize transfer conditions and verify that protein remains on neither the gel nor passes through the membrane. The supplied features do not establish one required transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A15258 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 SLC35D3 bands be quantified?
Quantitation · Quantify the same validated band across samples using exposures within the linear detection range. SLC35D3 is reported in vesicle, early endosome and endoplasmic reticulum membranes, so keep sample preparation and the fraction analyzed consistent across comparisons.
Why might SLC35D3 appear at 68 kDa instead of 44.2 kDa?
Interpretation · 44.2 kDa is the predicted mass, while 68 kDa is the supplied apparent band. SLC35D3 is a multi-pass membrane protein, but the supplied features do not establish why it migrates at 68 kDa. Compare the band with appropriate controls before assigning it to SLC35D3.
The record lists one isoform and no annotated glycosylation or modified residues. Those features do not identify an unexpected band. Compare it with the supplied 68 kDa apparent band and use specificity controls before assigning it to SLC35D3.
The record lists one isoform and no annotated glycosylation or modified residues. Those features do not identify an unexpected band. Compare it with the supplied 68 kDa apparent band and use specificity controls before assigning it to SLC35D3.
Boster reagents
SLC35D3 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-SLC35D3 Antibody
Cat # A15258
The catalog reports A15258, an anti-SLC35D3 antibody with reported human and mouse reactivity. Its WB image shows SLC35D3 analysis in HeLa cell lysate at 1 μg/mL; the supplied evidence does not show a mouse WB sample.
Which to pick: A15258 is the only listed option and has a WB image using HeLa cell lysate at 1 μg/mL. For mouse samples, reactivity is listed, but no mouse WB example is supplied.
Source: BosterBio SLC35D3 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q5M8T2.
- Human Protein Atlas. SLC35D3 tissue expression.
- PMC8373376 — target-verified WB comparison
- PMC10667520 — target-verified WB comparison