PTCH2 / Protein patched homolog 2 · Western blot design guide

Design a Western Blot for PTCH2

Source-linked PTCH2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PTCH2 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 PTCH2: expected band ~130.5 kDa, hero antibody A06375-1, catalog values and labelled standard workflow; separate PMC comparisons on the guide — Printable **PTCH2** Western blot protocol sheet — expected band ~130.5 kDa, antibody A06375-1, controls and PMC citations. Open the full PTCH2 WB guide →

Navigate to: Protocols Band Controls Antibodies

PTCH2 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	~130.5 kDa
	Observed band	~150 kDa
	Gel	5–20% (catalog A06375-1)
	Negative control ⓘ	Suggested KO / knockdown lysate

Important caveats

Reasons your observed band may differ from the expected size.

	PTM	Glycosylated
	Caveat	Modification-state controls
	Gene-set association	MSigDB C7 membership
	Isoform	2 isoform(s)

Section 1

Source-Linked PTCH2 Western Blot Protocol Options

The A06375-1 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 THP-1 (catalog A06375-1)
Gel %	5–20% (catalog A06375-1)
Load	50ug; reducing conditions (catalog A06375-1)
Transfer	a Nitrocellulose membrane at 150mA for 50-90 minutes (catalog A06375-1)
Membrane	Nitrocellulose membrane (catalog A06375-1)
Blocking	5% Non-fat Milk/ TBS for 1.5 hour at RT (catalog A06375-1)
Primary antibody	A06375-1 · 0.5 μg/mL (catalog A06375-1)
Primary incubation	overnight at 4°C (catalog A06375-1)
Secondary antibody	goat anti-rabbit IgG-HRP, 1:5000 (catalog A06375-1)
Secondary incubation	1.5 hour at RT (catalog A06375-1)
Wash	TBS-0.1%Tween 3 times with 5 minutes each (catalog A06375-1)
Detection	ECL (catalog A06375-1)

Section 2

What Is the Expected PTCH2 Western Blot Band Size?

PTCH2 is predicted at 130.5 kDa, while an empirical reducing-condition band appears at ~150 kDa; the cause of the difference is not established.

What am I looking at on my blot?
Band at ~150 kDa	Empirical PTCH2 band observed under reducing conditions; confirm identity with antibody controls.
Band near 130.5 kDa	Migration near the sequence-predicted mass; confirm PTCH2 identity.
Bands at different positions	Could reflect isoforms 1 and 2, although distinct migration is unestablished.
Broader or higher signal	Could reflect N-linked glycosylation at Asn370 and Asn812; a visible effect is unestablished.

💡Expected PTCH2 appearancePTCH2 has a predicted mass of 130.5 kDa and an observed band at ~150 kDa under reducing conditions; the cause of the difference is unestablished, so confirm band identity with appropriate controls.

How each factor affects band size
UniProt predicted mass	130.5 kDa by sequence; the empirical PTCH2 band is ~150 kDa under reducing conditions.
N-linked glycosylation at Asn370	May affect apparent size, but its contribution to the observed band is unestablished.
N-linked glycosylation at Asn812	May affect apparent size, but its contribution to the observed band is unestablished.
Splice isoforms 1 and 2	May differ in size; their relative migration is not supplied.

Why is my band missing or off?
Situation	Likely cause	Next action
No band in lysate	PTCH2 is a multi-pass membrane protein and may be poorly recovered.	Check membrane-protein extraction and include a positive control.
Band higher than expected	The ~150 kDa empirical band exceeds the 130.5 kDa predicted mass; the cause is unestablished.	Compare with the reported PTCH2 control band and test whether deglycosylation changes migration.
Band lower than expected	An alternative isoform is possible, but its migration is unknown.	Confirm identity with an independent antibody or PTCH2 depletion.
Broad smear instead of sharp band	Variable N-linked glycosylation is possible but unproven.	Compare untreated and deglycosylated samples with an identity control.
Multiple bands	Isoforms 1 and 2 are annotated, but distinct bands are not established.	Check which bands respond to PTCH2 depletion.
Weak or no signal	Recovery of this multi-pass membrane protein may be low.	Check extraction and loading with a membrane-protein positive control.

Source: UniProt Q9Y6C5 Human Protein Atlas product datasheet (A06375-1)

Experiment evidence

Western blot images showing PTCH2 band

A06375-1 · PTCH2

PA1859 · PTCH2

← swipe / scroll for more images →

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

Sample controls for PTCH2 Western blot

🧪For positive controls for PTCH2 in Western blot, you can use no HPA-supported tissue or cell sample because none 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: PTCH2 is a multi-pass membrane protein, and the supplied HPA data cannot identify a positive tissue.

HPA tissue expression evidence for PTCH2

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 PTCH2 Western Blot Tips

Deeper troubleshooting and optimisation questions for PTCH2, answered from its protein features.

How should PTCH2 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 PTCH2 isoforms produce different bands?

Isoforms · UniProt lists isoforms 1 and 2. In isoform 2, residues 1143–1203 of the canonical sequence are replaced by PEEI, so their sizes may differ. Check whether the antibody recognizes a region shared by both isoforms before interpreting separate bands. These are UniProt coordinates; antibody documentation may use different numbering.

Which PTCH2 modifications matter when interpreting a band?

PTM · UniProt lists N-linked glycosylation at asparagines 370 and 812. If testing whether glycosylation affects migration, compare matched samples with and without an appropriate deglycosylation treatment. A listed site does not guarantee a visible shift. Positions 370 and 812 use UniProt numbering.

Does this guide establish induction of PTCH2?

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 PTCH2 Western blot?

Transfer · PTCH2 is a 1203-residue multipass membrane protein with a predicted mass of 130.5 kDa. Use transfer conditions validated for proteins around the supplied ~150 kDa observed band, and check transfer efficiency in that region. The UniProt features do not specify a uniquely suitable transfer method.

How should blocking be optimized for PTCH2?

Blocking · PTCH2 is a multipass membrane glycoprotein, but its listed features do not identify a preferred blocker. Compare blocking conditions using matched samples and keep the condition that gives a clear band in the expected region with low background.

How can PTCH2 bands be quantified across samples?

Quantitation · Use the same sample preparation and detection conditions across lanes, and quantify bands within the assay’s linear range. If comparing isoforms, confirm that the antibody recognizes a shared region; isoform 2 replaces canonical residues 1143–1203 with PEEI. Report which band was measured rather than combining distinct bands without validation.

Why might PTCH2 appear near 150 kDa instead of 130.5 kDa?

Interpretation · 130.5 kDa is the predicted mass, while ~150 kDa is the supplied observed band. PTCH2 has N-linked glycosylation sites at UniProt positions 370 and 812, but their presence alone does not establish the cause or size of the difference. Compare the band with an appropriate molecular weight marker and avoid assigning the difference solely to glycosylation.

Check whether a band could reflect the documented isoform 2 sequence change or glycosylation at UniProt positions 370 and 812. Neither feature alone identifies an unexpected band. Compare reproducibility across samples and verify that the antibody recognizes the relevant isoform region before assigning its identity.

Boster reagents

PTCH2 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 PTCH2 using anti-PTCH2 antibody (A06375-1). 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 THP-1 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-PTCH2 antigen affinity purified polyclonal antibody (Catalog # A06375-1) 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 PTCH2 at approximately 150KD. The expected band size for PTCH2 is at 150KD.

Anti-PTCH2 Antibody Picoband®

Cat # A06375-1

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Real WB data Anti-PTCH2 antibody, PA1859, Western blotting WB: HELA Cell Lysate

Anti-PTCH2 Antibody Picoband®

Cat # PA1859

View product →

Two anti-PTCH2 antibodies have reported Western blot images. A06375-1 shows a band near the expected 150 kDa in human THP-1 lysate under stated conditions; PA1859 shows a blot using HeLa lysate. The supplied evidence does not document mouse WB testing or independent validation.

Which to pick: For human THP-1 lysate, A06375-1 has the more detailed WB example. PA1859 lists human and mouse reactivity, but its supplied WB example uses HeLa lysate; consider it for mouse samples with that evidence limit in mind.

Source: BosterBio PTCH2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.

References

UniProt Consortium. UniProt entry Q9Y6C5.
Human Protein Atlas. PTCH2 tissue expression.
PMC7181751 — target-verified WB comparison