This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
SLC16A1 / Monocarboxylate transporter 1 · Western blot design guide
Design a Western Blot for SLC16A1
Real validated SLC16A1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SLC16A1 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
SLC16A1 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 | ~53.9 kDa | |
| Observed band | ~40 kDa | |
| Gel | 8% (catalog A02240-1) | |
| Positive control | Appendix (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 | |
| Caveat | Observed below predicted | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated SLC16A1 Western Blot Protocols
The A02240-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human U87, human HL-60, human K562 (catalog A02240-1) |
| Gel % | 8% (catalog A02240-1) |
| Load | 30 ug; reducing conditions (catalog A02240-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A02240-1) |
| Membrane | nitrocellulose membrane (catalog A02240-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A02240-1) |
| Primary antibody | A02240-1 · 1:1000 (catalog A02240-1) |
| Primary incubation | overnight at 4°C (catalog A02240-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A02240-1) |
| Secondary incubation | 1.5 hour at RT (catalog A02240-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A02240-1) |
| Detection | ECL (catalog A02240-1) |
Section 2
What Is the Expected SLC16A1 Western Blot Band Size?
SLC16A1 is predicted at 53.9 kDa, but antibody QC shows approximately 40 kDa; the cause of the difference is not established.
| Band near 40 kDa | Empirical SLC16A1 band in antibody QC blots; confirm identity with controls |
| Band near 53.9 kDa | Near the sequence-predicted mass, without a demonstrated band at this position |
| Multiple bands at different positions | Isoforms 1 and 2 are possible contributors; their migration is unknown |
| Faint or absent band in whole-cell lysate | Membrane-associated SLC16A1 may be poorly recovered |
💡Expected SLC16A1 appearanceSLC16A1 has a predicted mass of 53.9 kDa, while antibody QC shows a band near 40 kDa; the cause of the difference is unestablished, so confirm identity with ordinary specificity controls.
| Predicted sequence mass | 53.9 kDa is the calculated mass, while the observed band is near 40 kDa |
| Splice isoform 1 | Its apparent size relative to isoform 2 is not supplied |
| Splice isoform 2 | Its apparent size relative to isoform 1 is not supplied |
| Alternative splicing of isoforms 1 and 2 | Could affect band position, but distinct migration has not been demonstrated |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated SLC16A1 may be poorly recovered | Check membrane protein recovery and use a positive-control lysate |
| Band higher than expected | Band identity or isoform contribution is uncertain | Compare with a positive control and verify specificity by SLC16A1 depletion |
| Band lower than expected | The observed 40 kDa band differs from the 53.9 kDa prediction for an unestablished reason | Compare with antibody QC conditions and verify specificity by SLC16A1 depletion |
| Multiple bands | Isoforms 1 and 2 are possible contributors, but their migration is unknown | Use isoform-specific controls to assign bands |
| Weak or no signal | Recovery of the multi-pass membrane protein may be low | Check solubilization and enrichment of the membrane fraction |
Experiment evidence
Western blot images showing SLC16A1 band
A02240-1 · SLC16A1
M02240 · SLC16A1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SLC16A1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SLC16A1 in Western blot, you can use appendix tissue, which HPA rates High.
Positive control: Appendix (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: As a multi-pass membrane protein, SLC16A1 may be easier to detect in a membrane-enriched lysate.
HPA tissue expression evidence for SLC16A1
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 |
|---|---|---|---|---|
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | endothelial cells | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | 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 → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cervix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SLC16A1 Western Blot Tips
Deeper troubleshooting and optimisation questions for SLC16A1, answered from its protein features.
How should SLC16A1 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 isoform 2 affect the Western blot pattern?
Isoforms · Isoform 2 replaces canonical residues 411–500 with a 20-residue sequence, making it shorter than isoform 1. An isoform-specific band is possible, but the supplied features do not identify the observed ~40 kDa band. Check which isoform the antibody recognizes before assigning bands.
That depends on its epitope. Isoform 2 replaces canonical residues 411–500 with a different 20-residue sequence. An antibody against the replaced canonical region may miss isoform 2; an antibody against a shared region could detect both. Check the antibody's mapped epitope against the isoform sequences.
That depends on its epitope. Isoform 2 replaces canonical residues 411–500 with a different 20-residue sequence. An antibody against the replaced canonical region may miss isoform 2; an antibody against a shared region could detect both. Check the antibody's mapped epitope against the isoform sequences.
Which SLC16A1 phosphorylation sites matter when interpreting bands?
PTM · UniProt lists phosphoserines at 210, 213, 461, 467, 483 and 498, and phosphothreonines at 231 and 466. These are UniProt canonical-sequence coordinates; antibody or paper numbering may differ. Phosphorylation at these sites does not by itself establish a visible band shift. The sites at 461–498 lie in the region replaced in isoform 2.
Does this guide establish induction of SLC16A1?
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 SLC16A1 Western blot?
Transfer · SLC16A1 is a multi-pass membrane protein, but the supplied features do not establish a preferred transfer method. Evaluate transfer with a total-protein stain and compare membrane retention with protein left in the gel, particularly around the observed ~40 kDa region.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A02240-1 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 SLC16A1 bands be quantified?
Quantitation · Measure the validated SLC16A1 band within the assay's linear signal range and normalize to a suitable loading measure. Because SLC16A1 is a cell-membrane protein whose targeting involves EMB or BSG isoform 2, keep the sampled fraction consistent when comparing conditions.
Why might SLC16A1 appear near 40 kDa instead of 53.9 kDa?
Interpretation · The supplied blots show an apparent band near 40 kDa, while the canonical sequence has a predicted mass of 53.9 kDa. SLC16A1 is a multi-pass membrane protein and has a shorter isoform 2, but these features alone do not establish why the band migrates there. Confirm band identity with an appropriate specificity control.
Consider the two listed isoforms, antibody cross-reactivity, and sample-dependent migration of this multi-pass membrane protein. The listed phosphorylation sites do not prove that any additional band is phosphorylated SLC16A1. Use a specificity control and consider whether the antibody epitope is shared by both isoforms.
Consider the two listed isoforms, antibody cross-reactivity, and sample-dependent migration of this multi-pass membrane protein. The listed phosphorylation sites do not prove that any additional band is phosphorylated SLC16A1. Use a specificity control and consider whether the antibody epitope is shared by both isoforms.
Boster reagents
SLC16A1 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-MCT1/SLC16A1 Antibody
Cat # A02240-1
Real WB data
Anti-MCT1 Rabbit Monoclonal Antibody
Cat # M02240
Two the supplier anti-SLC16A1 antibodies have Western blot images using human whole cell lysates. A02240-1 shows a band near 40 kDa; M02240 shows one near 50 kDa, against a stated expected size of 54 kDa. The supplied images do not demonstrate mouse or rat samples.
Which to pick: For human samples, both have WB images; M02240 shows a band closer to the stated 54 kDa size. A02240-1 lists human, mouse, and rat reactivity, but its supplied WB image shows only human samples. Choose based on species and the reported band sizes.
Source: BosterBio SLC16A1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P53985.
- Human Protein Atlas. SLC16A1 tissue expression.
- PMC9249469 — target-verified WB comparison
- PMC11094506 — target-verified WB comparison
- PMC4694819 — target-verified WB comparison