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- Table of Contents
SLC25A1 / Tricarboxylate transport protein, mitochondrial · Western blot design guide
Design a Western Blot for SLC25A1
Source-linked SLC25A1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SLC25A1 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
SLC25A1 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 | ~34 kDa | |
| Observed band | ~30–34 kDa | |
| Gel | 5–20% (catalog A05995-1) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Cleaved | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked SLC25A1 Western Blot Protocol Options
The A05995-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human Jurkat, human CCRF-CEM, rat brain, mouse brain (catalog A05995-1) |
| Gel % | 5–20% (catalog A05995-1) |
| Load | 50ug; reducing conditions (catalog A05995-1) |
| Transfer | a Nitrocellulose membrane at 150mA for 50-90 minutes (catalog A05995-1) |
| Membrane | Nitrocellulose membrane (catalog A05995-1) |
| Blocking | 5% Non-fat Milk/ TBS for 1.5 hour at RT (catalog A05995-1) |
| Primary antibody | A05995-1 · 0.5 μg/mL (catalog A05995-1) |
| Primary incubation | overnight at 4°C (catalog A05995-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A05995-1) |
| Secondary incubation | 1.5 hour at RT (catalog A05995-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A05995-1) |
| Detection | ECL (catalog A05995-1) |
Section 2
What Is the Expected SLC25A1 Western Blot Band Size?
SLC25A1 is predicted at 34 kDa and observed at ~30–34 kDa; the cause of any difference is not established.
| Band at ~30–34 kDa | Matches the empirical SLC25A1 band; the predicted precursor mass is 34 kDa |
| Band slightly below the precursor | Could reflect removal of the annotated residues 1–13 propeptide; its migration has not been established |
| Faint or absent band in whole-cell lysate | SLC25A1 resides in the mitochondrial inner membrane and may be poorly recovered |
| Single band without a doubled-mass partner | Consistent with the annotated monomer |
💡Expected SLC25A1 appearanceSLC25A1 is predicted at 34 kDa and has an empirical band at ~30–34 kDa; confirm band identity with an appropriate specificity control.
| Predicted precursor mass | 34 kDa from the UniProt sequence; the empirical band is ~30–34 kDa |
| Residues 1–13 propeptide | Cleavage could make the mature protein slightly smaller than the precursor; its contribution to the observed band is unproven |
| Phosphoserine 156 | No apparent size change is established |
| Mitochondrial inner membrane localization | No size shift is established; recovery from lysate may affect band detection |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Poor recovery of the mitochondrial inner membrane protein | Enrich mitochondria and optimize membrane solubilization |
| Band higher than expected | Identity of the higher band is unestablished; SLC25A1 is annotated as a monomer | Check specificity with SLC25A1 knockdown and a molecular weight marker |
| Band lower than expected | Propeptide cleavage is possible, but the cause of a lower band is unproven | Check specificity and compare precursor and mature N termini if needed |
| Multiple bands | Band identities and any processing difference are unestablished | Use SLC25A1 knockdown to identify specific bands |
| Weak or no signal | The inner membrane protein may be poorly extracted | Check extraction and enrich the mitochondrial fraction |
| Fragments below expected size | Protein degradation is possible; fragment identities are unestablished | Use protease inhibitors during preparation and verify specificity |
Experiment evidence
Western blot images showing SLC25A1 band
A05995-1 · SLC25A1
A05995-2 · SLC25A1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SLC25A1 Western blot
🧪For positive controls for SLC25A1 in Western blot, you can use a verified SLC25A1-expressing mitochondrial lysate; no HPA positive tissue or cell line 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 expression data, so a positive sample must be verified and a knockdown or KO is needed for a reliable negative control.
HPA tissue expression evidence for SLC25A1
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 SLC25A1 Western Blot Tips
Deeper troubleshooting and optimisation questions for SLC25A1, answered from its protein features.
How should SLC25A1 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 SLC25A1 isoforms explain multiple bands?
Isoforms · UniProt lists one isoform and no alternative sequence. Do not assign additional bands to annotated isoforms without separate evidence.
How should Ser156 phosphorylation affect band interpretation?
PTM · UniProt annotates phosphoserine at position 156 in its own numbering. It does not specify an enzyme or induction condition. The annotation alone does not establish a visible shift; a phosphatase comparison can test whether an observed mobility difference depends on phosphorylation.
Does this guide establish induction of SLC25A1?
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 SLC25A1 Western blot?
Transfer · Optimize transfer for the ~30–34 kDa band and check whether protein remains in the gel. SLC25A1 is a multi-pass inner-membrane protein, so verify recovery under the chosen transfer conditions rather than assuming it transferred efficiently.
How should blocking be optimized for SLC25A1?
Blocking · Compare blocking conditions using the ~30–34 kDa signal and background as readouts. SLC25A1 is a multi-pass mitochondrial inner-membrane protein; its listed features do not establish that one blocker will work best.
How can SLC25A1 be quantified across samples?
Quantitation · Keep sample preparation consistent, particularly mitochondrial or membrane enrichment, because SLC25A1 resides in the mitochondrial inner membrane. Quantify the ~30–34 kDa band within the linear signal range and use a consistent loading normalization method.
Does a ~30–34 kDa band match the predicted mass?
Interpretation · Yes. SLC25A1 has a predicted mass of 34 kDa, and the reported apparent band is ~30–34 kDa. UniProt annotates a propeptide at residues 1..13, but that feature alone does not establish cleavage or explain any mobility difference.
UniProt describes SLC25A1 as a monomer, lists one isoform, and lists no glycosylation sites. These features do not support assigning higher bands to annotated isoforms, glycans, or an obligate oligomer. Verify band specificity experimentally before assigning an identity.
UniProt describes SLC25A1 as a monomer, lists one isoform, and lists no glycosylation sites. These features do not support assigning higher bands to annotated isoforms, glycans, or an obligate oligomer. Verify band specificity experimentally before assigning an identity.
Boster reagents
SLC25A1 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-Slc25a1 Picoband® Antibody
Cat # A05995-1
Real WB data
Anti-Slc25a1 Antibody Picoband®
Cat # A05995-2
Both listed anti-SLC25A1 rabbit polyclonal antibodies have Western blot images from human, rat, and mouse samples. Reported bands are approximately 30–34 kDa for A05995-1 and 34 kDa for A05995-2. The supplied evidence is limited to product images and captions.
Which to pick: Both list human, mouse, and rat reactivity. Choose A05995-1 for its reported HeLa, Jurkat, CCRF-CEM, or brain lysate examples; choose A05995-2 for HepG2, Jurkat, K562, A549, rat liver or RH35, or mouse HEPA1-6 examples. Match your sample to the reported conditions.
Source: BosterBio SLC25A1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P53007.
- Human Protein Atlas. SLC25A1 tissue expression.
- PMC11753767 — target-verified WB comparison
- PMC8627508 — target-verified WB comparison
- PMC10840918 — target-verified WB comparison