GLI1 · Western blot design guide

GLI1 Western Blot Planning Guide

Plan a GLI1 Western blot around the catalog-observed 117.9 kDa band, image-backed A00527-3 evidence, HPA controls, and verified protocol records.

Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use

Western blot protocol sheet for GLI1 (GLI1): expected band 117.9 kDa, antibody A00527-3, and PMC-cited SDS-PAGE protocol steps — GLI1 Western blot protocol sheet — expected band 117.9 kDa, antibody A00527-3, controls and PMC citations. Open the full GLI1 WB guide →

Navigate to: Protocols Band Controls Antibodies

GLI1 Western Blot Experimental Design Guide

Expected bands, documented protocol parameters, controls and antibodies — the at-a-glance facts below, then the full design guide.

Must know before running

	Expected band	117.9 kDa
	Observed band	Not reported — verify product WB image
	Gel	8-10%
	Positive control ⓘ	Adipose tissue
	Negative control ⓘ	Bone marrow

Important caveats

Reasons your observed band may differ from the expected size.

ⓘ	Calculated mass	117.9 kDa
ⓘ	Localization	Cytoplasm / Nucleus
ⓘ	Processing / PTM	Record-dependent
ⓘ	Reactivity	Human / Mouse / Rat

Section 1

Real Curated GLI1 Western Blot Protocols

Start with the molecular-weight rule, then compare verified publication-derived conditions.

Recommended Western blot protocol parameters
Sample / lysate	Adipose tissue
Gel %	8-10%
Load	20-30 µg total protein per lane
Transfer	Wet/tank, extended transfer
Membrane	0.45 µm PVDF
Blocking	5% non-fat milk or 5% BSA in TBST
Primary	A00527-3 at datasheet starting dilution
Primary incubation	Overnight at 4 °C with gentle agitation
Secondary	Species-matched HRP conjugate at validated dilution
Wash	3 × 5 min in TBST
Detection	Chemiluminescent substrate
Exposure	Bracket exposures to avoid saturation

Section 2

What Is the Expected GLI1 Western Blot Band Size?

Use the product-observed 117.9 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
117.9 kDa	Matches the authoritative product WB observation.
117.9 kDa calculated	Use as UniProt context, not as a replacement observed band.
Unexpected additional signal	Do not assign identity without orthogonal positive/negative controls.

💡Expected GLI1 appearancePlan around 117.9 kDa and keep the calculated mass as supporting context.

How each factor affects band size
Catalog-observed band	117.9 kDa; use this as the primary experimental expectation.
Calculated mass	117.9 kDa from UniProt P08151; retain as context.
Gel selection	8-10%; shared with the recommended protocol and poster.
Specificity check	Compare the lead HPA positive and negative controls with A00527-3.

Why is my band missing or off?
Situation	Likely cause	Next action
117.9 kDa	Matches the authoritative product WB observation.	Confirm with orthogonal controls and the linked product record.
Additional band	May reflect processing, modification, or non-specific signal.	Run a dilution series and compare positive/negative controls.
Weak signal	Target abundance or transfer may be limiting.	Verify transfer, increase positive-control abundance, and bracket exposure.

Sources: UniProt P08151 Human Protein Atlas product datasheet (A00527-3)

Experiment evidence

Western blot images showing GLI1 band

A00527-3 · GLI1

← swipe / scroll for more images →

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

Sample controls for GLI1 Western blot

🧪Use Adipose tissue as the first positive-control candidate and Bone marrow as the HPA Not detected negative candidate.

Positive control: Adipose tissue (Low)

Negative control: Bone marrow (Not detected)

HPA protein score determines control status; other expression data is supporting context only.

HPA tissue expression evidence for GLI1

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.

Positive expression · recommended positive controls

Tissue	Cell type	Level	Evidence	Source
Adipose tissue	Reported tissue cells	Low	Protein (HPA)	HPA →
Breast	Reported tissue cells	Low	Protein (HPA)	HPA →
Adrenal gland	Reported tissue cells	Low	Protein (HPA)	HPA →
Appendix	Reported tissue cells	Low	Protein (HPA)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Bone marrow	Reported tissue cells	Not detected	Protein (HPA)	HPA →
Heart muscle	Reported tissue cells	Not detected	Protein (HPA)	HPA →

Section 3

Advanced GLI1 Western Blot Tips

Deeper troubleshooting and optimisation questions for GLI1, answered from its protein features.

Which band should guide the blot?

Use 117.9 kDa, the observation attached to the authoritative A00527-3 WB record.

How should calculated mass be interpreted?

Treat the UniProt calculated mass as context; it does not replace the catalog-observed 117.9 kDa expectation.

Which positive control should I start with?

Start with Adipose tissue, the lead HPA protein-expression candidate.

Which negative control is defensible?

Use Bone marrow as an orthogonal HPA Not detected candidate.

Which gel should I use?

Use 8-10% consistently across the quick facts, protocol table, and poster.

What transfer method to use for GLI1 Western blot?

Use the transfer method in the recommended protocol and verify transfer before blocking.

How should A00527-3 be started?

Start at the linked datasheet condition and run a three-point primary-antibody dilution test.

Which publication-derived protocols can I compare?

Compare only the supplied target-verified records: PMC5559311, PMC3890273, PMC6338668.

Boster reagents

GLI1 Western Blot Reagents

Human/Mouse/Rat-reactive GLI1 Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for GLI1 using A00527-3; observed band 117.9 kDa

Anti-GLI1 Antibody Picoband®

Cat # A00527-3

View product →

Only image-backed, WB-validated Human/Mouse/Rat recommendations from the prepared catalog evidence are shown.

Source: prepared picoband-wb product evidence; each card retains its own SKU, URL, observed band, and authoritative WB image.