MESP1 / Mesoderm posterior protein 1 · Western blot design guide

Design a Western Blot for MESP1

Real validated MESP1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MESP1 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

Western blot protocol sheet for MESP1: expected band ~28.5 kDa, hero antibody A07301-2, catalog values and labelled standard workflow; separate PMC comparisons on the guide — Printable **MESP1** Western blot protocol sheet — expected band ~28.5 kDa, antibody A07301-2, controls and PMC citations. Open the full MESP1 WB guide →

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MESP1 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	~28.5 kDa
	Observed band	~29 kDa
	Gel	5–20% (catalog A07301-2)
	Negative control ⓘ	Suggested KO / knockdown lysate

Important caveats

Reasons your observed band may differ from the expected size.

	PTM	—
	Caveat	—
	Gene-set association	MSigDB C7 membership
	Isoform	1 isoform(s)

Section 1

Real Curated MESP1 Western Blot Protocols

The A07301-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.

Recommended Western blot protocol parameters
Sample / lysate	human A549, human Hela, human HEK293 (catalog A07301-2)
Gel %	5–20% (catalog A07301-2)
Load	50ug; reducing conditions (catalog A07301-2)
Transfer	a Nitrocellulose membrane at 150mA for 50-90 minutes (catalog A07301-2)
Membrane	Nitrocellulose membrane (catalog A07301-2)
Blocking	5% Non-fat Milk/ TBS for 1.5 hour at RT (catalog A07301-2)
Primary antibody	A07301-2 · 0.5 μg/mL (catalog A07301-2)
Primary incubation	overnight at 4°C (catalog A07301-2)
Secondary antibody	goat anti-rabbit IgG-HRP, 1:5000 (catalog A07301-2)
Secondary incubation	1.5 hour at RT (catalog A07301-2)
Wash	TBS-0.1%Tween 3 times with 5 minutes each (catalog A07301-2)
Detection	ECL (catalog A07301-2)

Section 2

What Is the Expected MESP1 Western Blot Band Size?

MESP1 is predicted at 28.5 kDa and observed at ~29 kDa; the cause of the small difference is not established.

What am I looking at on my blot?
Single band at ~29 kDa	Consistent with the reported MESP1 band; confirm identity with controls
Band near 28.5 kDa	Near the predicted MESP1 mass; confirm identity with controls
Band above ~29 kDa	Its identity and higher position are unexplained by the supplied features
Band below ~29 kDa	Its identity and lower position are unexplained by the supplied features

💡Expected MESP1 appearanceMESP1 has a predicted mass of 28.5 kDa and an empirical band at ~29 kDa; use band-identity controls to confirm the signal.

How each factor affects band size
UniProt predicted mass of 28.5 kDa	Provides the calculated size reference
Predicted MESP1 protein mass	Places the expected band near 29 kDa
Predicted mass compared with the empirical band	Differs slightly from the observed ~29 kDa position
Predicted mass as a band-identity reference	Does not establish the identity of bands at other sizes

Why is my band missing or off?
Situation	Likely cause	Next action
No band in lysate	MESP1 may be below detection in the tested sample	Check a nuclear-enriched sample and a positive control
Band higher than expected	The supplied features do not explain the higher position	Check markers and verify band identity with an independent antibody
Band lower than expected	The supplied features do not establish a smaller processed form	Verify band identity with an independent antibody
Multiple bands	The supplied features do not establish multiple MESP1 isoforms	Compare bands with a positive control and an independent antibody
Weak or no signal	MESP1 may be below detection in the tested sample	Check nuclear enrichment and assay performance with a positive control

Source: UniProt Q9BRJ9 Human Protein Atlas product datasheet (A07301-2)

Experiment evidence

Western blot images showing MESP1 band

A07301-2 · MESP1

← swipe / scroll for more images →

Real WB data behind the band-size expectations in this section.

Sample controls for MESP1 Western blot

🧪For positive controls for MESP1 in Western blot, you can use no HPA-supported sample because no positive tissue or cell is reported.

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 such as Ponceau alongside the samples.

⚠️Feasibility: HPA provides no expression data, so suitable tissue controls cannot be identified.

HPA tissue expression evidence for MESP1

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 MESP1 Western Blot Tips

Deeper troubleshooting and optimisation questions for MESP1, answered from its protein features.

How should MESP1 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.

Should MESP1 isoforms produce multiple bands?

Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no isoform-based explanation for multiple bands.

Could annotated modifications explain a MESP1 band shift?

PTM · No modified residues or glycosylation sites are listed. The supplied features do not support assigning a shifted band to a specific modification, though they cannot rule out unannotated modifications.

Does this guide establish induction of MESP1?

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 MESP1 Western blot?

Transfer · MESP1 is predicted to be 28.5 kDa. The supplied features do not specify a transfer method; check transfer efficiency around 29 kDa when setting up the blot.

How should blocking be optimized?

Blocking · Standard workflow guidance: follow the A07301-2 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 MESP1 be quantified in nuclear samples?

Quantitation · MESP1 is annotated as nuclear. Use consistently prepared nuclear samples and normalize the approximately 29 kDa signal against a suitable nuclear loading measure.

Why is the MESP1 band near its predicted mass?

Interpretation · MESP1 is predicted at 28.5 kDa, close to the reported apparent band at approximately 29 kDa. This agreement does not establish the band's identity; confirm it with an appropriate specificity control.

The record lists one isoform, no alternative sequence, and no annotated modified residues or glycosylation sites. These features do not identify the cause of an unexpected band. Compare it with the approximately 29 kDa band and use a specificity control before assigning it to MESP1.

Boster reagents

MESP1 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

Western blot analysis of MESP1 using anti-MESP1 antibody (A07301-2). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: human A549 whole cell lysates, Lane 2: human Hela whole cell lysates, Lane 3: human HEK293 whole cell lysates, Lane 4: human Caco-2 whole cell lysates, Lane 5: human Raji whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-MESP1 antigen affinity purified polyclonal antibody (Catalog # A07301-2) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for MESP1 at approximately 29KD. The expected band size for MESP1 is at 29KD.

Anti-MESP1 Antibody Picoband®

Cat # A07301-2

View product →

A07301-2 is a rabbit polyclonal anti-MESP1 antibody with a Western blot image showing an approximately 29 kDa band in human A549, HeLa, HEK293, Caco-2, and Raji whole-cell lysates. The supplied evidence covers these samples under the reported conditions.

Which to pick: A07301-2 is the only listed option. It reports human reactivity and has a Western blot image from five human cell lines, using 50 µg lysate per lane and 0.5 µg/mL primary antibody.

Source: BosterBio MESP1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.

References

UniProt Consortium. UniProt entry Q9BRJ9.
Human Protein Atlas. MESP1 tissue expression.
PMC7295063 — target-verified WB comparison
PMC6921370 — target-verified WB comparison
PMC12229608 — target-verified WB comparison