LPAR1 / Lysophosphatidic acid receptor 1 · Western blot design guide

Design a Western Blot for LPAR1

Real validated LPAR1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-LPAR1 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 LPAR1: expected band ~41.1 kDa, hero antibody M02782, catalog values and labelled standard workflow; separate PMC comparisons on the guide — Printable **LPAR1** Western blot protocol sheet — expected band ~41.1 kDa, antibody M02782, controls and PMC citations. Open the full LPAR1 WB guide →

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LPAR1 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	~41.1 kDa
	Observed band	~39 kDa
	Gel	10% (catalog M02782)
	Negative control ⓘ	Suggested KO / knockdown lysate

Important caveats

Reasons your observed band may differ from the expected size.

	PTM	Glycosylated + Phosphorylated
	Caveat	Modification-state controls
	Gene-set association	MSigDB Hallmark membership
	Isoform	2 isoform(s)

Section 1

Real Curated LPAR1 Western Blot Protocols

The M02782 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 Hela, human Jurkat, human HepG2 (catalog M02782)
Gel %	10% (catalog M02782)
Load	30 ug; reducing conditions (catalog M02782)
Transfer	a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M02782)
Membrane	nitrocellulose membrane (catalog M02782)
Blocking	5% non-fat milk/TBS for 1.5 hour at RT (catalog M02782)
Primary antibody	M02782 · 1:500 (catalog M02782)
Primary incubation	overnight at 4°C (catalog M02782)
Secondary antibody	goat anti-rabbit IgG-HRP, 1:5000 (catalog M02782)
Secondary incubation	1.5 hour at RT (catalog M02782)
Wash	TBS-0.1%Tween 3 times with 5 minutes each (catalog M02782)
Detection	ECL (catalog M02782)

Section 2

What Is the Expected LPAR1 Western Blot Band Size?

LPAR1 is predicted at 41.1 kDa and observed near 39 kDa; the cause of the difference is not established.

What am I looking at on my blot?
Band near 39 kDa	Empirical LPAR1 band; confirm identity with appropriate controls
Band above the main band	N-linked glycosylation at Asn27 or Asn35 could affect migration; a shift is unproven
Several bands	Isoforms 1 and 2 or glycosylation may contribute; distinct bands are not established
Weak signal in a soluble fraction	LPAR1 is a multi-pass membrane protein and may be poorly recovered

💡Expected LPAR1 appearanceLPAR1 has a predicted mass of 41.1 kDa and an empirical band near 39 kDa; the reason for the difference is unestablished, so confirm band identity with appropriate controls.

How each factor affects band size
UniProt predicted mass	41.1 kDa is the sequence-based reference; an empirical band appears near 39 kDa
N-linked glycosylation at Asn27	May affect apparent size, but no migration effect is established
N-linked glycosylation at Asn35	May affect apparent size, but no migration effect is established
Splice isoforms 1 and 2	May differ in size; their masses and separation are not supplied

Why is my band missing or off?
Situation	Likely cause	Next action
No band in lysate	Membrane-associated LPAR1 may be poorly extracted	Check membrane protein recovery and a positive-control lysate
Band higher than expected	Glycosylation at Asn27 or Asn35 is possible, but its effect is unproven	Compare with a validated positive control and assess glycosylation if needed
Broad smear instead of sharp band	Variable glycosylation is possible but unconfirmed	Check sample quality and compare with a positive control
Multiple bands	Isoforms or glycosylation are possible; distinct migration is unconfirmed	Use band-identity controls to assess each band
Weak or no signal	Poor recovery of multi-pass membrane LPAR1 is possible	Check membrane extraction, loading and transfer
Band lower than expected	The observed 39 kDa band is below the 41.1 kDa prediction; the cause is unestablished	Compare with the empirical reference band and use band-identity controls

Source: UniProt Q92633 Human Protein Atlas product datasheet (M02782)

Experiment evidence

Western blot images showing LPAR1 band

M02782 · LPAR1

← swipe / scroll for more images →

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

Sample controls for LPAR1 Western blot

🧪For positive controls for LPAR1 in Western blot, you can use an HPA-IHC candidate positive sample once identified; none is supplied in the evidence.

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 needs validation and siRNA knockdown or a KO line is the practical negative control.

HPA tissue expression evidence for LPAR1

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 LPAR1 Western Blot Tips

Deeper troubleshooting and optimisation questions for LPAR1, answered from its protein features.

How should LPAR1 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 LPAR1 isoforms produce different Western blot bands?

Isoforms · UniProt lists isoforms 1 and 2. Isoform 2 replaces residues 1–15 with a different 15-residue sequence, so a large size difference is not predicted from this change alone. Check whether the antibody recognizes the changed N terminus before comparing isoform signals.

Which LPAR1 glycosylation sites matter when interpreting bands?

PTM · UniProt annotates N-linked glycosylation at Asn27 and Asn35. These are UniProt coordinates; antibody or paper numbering may differ. Glycosylation could affect migration, but the annotations do not establish a visible shift. Compare matched untreated and deglycosylated samples if testing that possibility.

UniProt annotates phosphoserine at Ser341 and phosphothreonine at Thr351, using UniProt coordinates. Verify the numbering convention before selecting a site-specific antibody. These annotations do not establish that phosphorylation changes the visible band position or increases after any particular treatment.

Does this guide establish induction of LPAR1?

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 LPAR1 Western blot?

Transfer · LPAR1 is a multi-pass membrane protein, with an apparent band near 39 kDa. Check transfer at that position with a total-protein stain and confirm that the membrane retains the target signal. The supplied features do not specify a preferred transfer method or settings.

How should blocking be optimized?

Blocking · Standard workflow guidance: follow the M02782 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 I quantify LPAR1 across samples?

Quantitation · LPAR1 is annotated at the cell surface, cell membrane and endosome. Keep sample preparation consistent across conditions, quantify the same verified band, and normalize to a suitable loading measure. If using a membrane-enriched fraction, compare like fractions across samples.

Why might LPAR1 migrate near 39 kDa instead of 41.1 kDa?

Interpretation · The supplied apparent band is about 39 kDa, versus a predicted mass of 41.1 kDa. LPAR1 is a multi-pass membrane protein with two annotated N-linked glycosylation sites, but these features alone do not explain the difference. Use the observed band as a reference and verify its identity independently.

First verify band identity. LPAR1 has two isoforms, two annotated N-linked glycosylation sites and three disulfide bonds, but those features do not by themselves identify an extra band. Compare antibody epitope coverage, matched deglycosylated samples and consistent reducing conditions as appropriate.

Boster reagents

LPAR1 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 LPAR1 using anti-LPAR1 antibody (M02782). Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human Hela whole cell lysates, Lane 2: human Jurkat whole cell lysates, Lane 3: human HepG2 whole cell lysates, Lane 4: human MCF-7 whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-LPAR1 antigen affinity purified monoclonal antibody (M02782) at 1:500 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 ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for LPAR1 at approximately 39 kDa. The expected band size for LPAR1 is at 39 kDa.

Anti-EDG2 LPAR1 Rabbit Monoclonal Antibody

Cat # M02782

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The catalog reports one anti-LPAR1 antibody, M02782, a rabbit monoclonal reported reactive with human samples. Its Western blot image shows an approximately 39 kDa band in HeLa, Jurkat, HepG2, and MCF-7 whole-cell lysates. No independent validation or publication evidence is supplied.

Which to pick: M02782 is the only listed option and has a Western blot image for four human cell lysates. The reported blot used 30 µg per lane and a 1:500 primary dilution. Performance in other samples is not established by the supplied evidence.

Source: BosterBio LPAR1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.

References

UniProt Consortium. UniProt entry Q92633.
Human Protein Atlas. LPAR1 tissue expression.
PMC6811758 — target-verified WB comparison
PMC12548558 — target-verified WB comparison
PMC12859871 — target-verified WB comparison