This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
MED1 / Mediator of RNA polymerase II transcription subunit 1 · Western blot design guide
Design a Western Blot for MED1
Real validated MED1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MED1 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
MED1 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 | ~168.5 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Adrenal gland (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 | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated MED1 Western Blot Protocols
The A00550-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | A549 (40ug), HEK293T (40ug), PC12 (40ug) (catalog A00550-1) |
| 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 | A00550-1; 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 MED1 Western Blot Band Size?
MED1 has a predicted mass of 168.5 kDa; isoforms and phosphorylation could affect migration, but no empirical band or feature-specific shift is established.
| Band near 168.5 kDa | consistent with the UniProt predicted mass; confirm identity with antibody controls |
| Several bands near the expected region | could reflect isoforms 1 and 2, although their migration is unknown |
| A close doublet | could reflect different phosphorylation states, but a visible shift is unproven |
| Stronger band in a nuclear fraction | consistent with MED1 nuclear localization |
💡Expected MED1 appearanceUniProt predicts MED1 at 168.5 kDa; no empirical band size is supplied, and isoforms or phosphorylation may affect migration, so confirm any candidate band with ordinary identity controls.
| UniProt predicted mass | places the sequence-based reference at 168.5 kDa |
| Isoform 1 | may migrate differently from isoform 2; its individual mass is unavailable |
| Isoform 2 | may migrate differently from isoform 1; its individual mass is unavailable |
| Alternative splicing | could change apparent size, but no resolvable band pattern is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | MED1 is nuclear and may be underrepresented in the sampled lysate | check a nuclear fraction and verify loading |
| Band higher than expected | migration may differ from the sequence-based prediction; the cause is unestablished | compare with a molecular weight marker and confirm identity by MED1 knockdown |
| Band lower than expected | an isoform or fragment is possible, but its size is not supplied | confirm identity by MED1 knockdown and compare antibodies to different epitopes |
| Multiple bands | isoforms 1 and 2 or phosphorylation states are possible; separation is unproven | compare nuclear fractions and test which bands diminish after MED1 knockdown |
| Weak or no signal | nuclear MED1 may be diluted in whole-cell lysate | enrich the nuclear fraction and verify sample loading |
Experiment evidence
Western blot images showing MED1 band
A00550-1 · MED1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MED1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for MED1 in Western blot, you can use adrenal gland tissue, which HPA scores as high expression.
Positive control: Adrenal gland (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: MED1 is nuclear, so a nuclear-enriched extract may improve detection; HPA lists adipose tissue as not detected.
HPA tissue expression evidence for MED1
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuropil | High | Protein (IHC) | HPA → |
| Colon | 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 → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | ciliated cells (cell body) | Low | Protein (IHC) | HPA → |
| Caudate | glial cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced MED1 Western Blot Tips
Deeper troubleshooting and optimisation questions for MED1, answered from its protein features.
How should MED1 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 MED1 isoforms produce different bands?
Isoforms · Yes. Isoform 2 replaces residues 548–556 and lacks residues 557–1581, so it is substantially shorter than the 1,581-residue canonical isoform. Check which isoform the antibody recognizes before assigning a lower band.
Choose a validated epitope within residues 1–547, which precede the isoform 2 sequence change. An epitope within canonical residues 557–1581 would be absent from isoform 2; residues 548–556 are replaced in isoform 2.
Choose a validated epitope within residues 1–547, which precede the isoform 2 sequence change. An epitope within canonical residues 557–1581 would be absent from isoform 2; residues 548–556 are replaced in isoform 2.
Which MED1 modifications matter when interpreting bands?
PTM · UniProt lists 26 modified residues, including phosphothreonine at positions 1032 and 1457 and N6-acetyllysine at 1177 and 1529. These are UniProt canonical-sequence coordinates; antibody or paper numbering may differ. Their presence alone does not establish a visible band shift.
Does this guide establish induction of MED1?
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 MED1?
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 A00550-1 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 MED1 bands be quantified across samples?
Quantitation · Use the same antibody and band definition across samples, accounting for whether the antibody recognizes both isoforms. MED1 is nuclear, and a phosphorylated subset may enter the nucleolus, so keep the sampled fraction consistent. Do not combine distinct bands as one species without establishing their identities.
Why might MED1 migrate differently from its predicted mass?
Interpretation · Canonical MED1 has a predicted mass of 168.5 kDa and 26 listed modified residues. The supplied features do not establish an apparent Western blot mass or show that modification causes a visible shift. Compare bands with a molecular weight marker and avoid assigning identity from size alone.
UniProt assigns phosphorylation of MED1 Thr1032 and Thr1457 to MAPK1 or MAPK3 and notes that a subset may enter the nucleolus after phosphorylation by either enzyme. If comparing conditions with differing MAPK activity, consider nuclear fraction composition when interpreting signal. The features specify no inducing treatment or expected band change.
Consider isoform 2, which lacks canonical residues 557–1581, but verify that the antibody can recognize it. The supplied features provide no observed band position and do not identify any particular lower band. Check epitope location and compare the signal across suitable samples before assigning it to MED1.
UniProt assigns phosphorylation of MED1 Thr1032 and Thr1457 to MAPK1 or MAPK3 and notes that a subset may enter the nucleolus after phosphorylation by either enzyme. If comparing conditions with differing MAPK activity, consider nuclear fraction composition when interpreting signal. The features specify no inducing treatment or expected band change.
Consider isoform 2, which lacks canonical residues 557–1581, but verify that the antibody can recognize it. The supplied features provide no observed band position and do not identify any particular lower band. Check epitope location and compare the signal across suitable samples before assigning it to MED1.
Boster reagents
MED1 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-TRAP220 (P697) MED1 Antibody
Cat # A00550-1
The catalog reports A00550-1 for MED1, with reported Human, Mouse, and Rat reactivity. Its WB image shows A549, HEK293T, PC12, and CT26 whole-cell lysates at 40 µg per lane and a 1:500 antibody dilution. The supplied evidence covers these tested conditions.
Which to pick: A00550-1 is the only listed option and has a WB image. Its reported reactivity and shown lysates can guide selection; validate performance for your own samples and conditions.
Source: BosterBio MED1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q15648.
- Human Protein Atlas. MED1 tissue expression.
- PMC8050805 — target-verified WB comparison
- PMC4388278 — target-verified WB comparison