GRB14 · Western blot design guide

GRB14 Western Blot Planning Guide

Plan a GRB14 Western blot around the catalog-observed 61 kDa band, image-backed A04177-2 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 GRB14 (GRB14): expected band 61 kDa, antibody A04177-2, and PMC-cited SDS-PAGE protocol steps

Band Protocols Controls Antibodies

GRB14 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	61 kDa
	Observed band	Not reported — verify product WB image
	Gel	12-15%
	Positive control ⓘ	Caudate
	Negative control ⓘ	Appendix

Important caveats

Reasons your observed band may differ from the expected size.

ⓘ	Calculated mass	61 kDa
ⓘ	Localization	Cytoplasm / Endosome membrane
ⓘ	Processing / PTM	Record-dependent
ⓘ	Reactivity	Human

Sample controls for GRB14 Western blot

🧪Use Caudate as the first positive-control candidate and Appendix as the HPA Not detected negative candidate.

Positive control: Caudate (Medium)

Negative control: Appendix (Not detected)

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

HPA tissue expression evidence for GRB14

Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart on each tissue page — click any row's HPA link to view the source.

Positive expression · recommended positive controls

Tissue	Cell type	Level	Evidence	Source
Caudate	Reported tissue cells	Medium	Protein (HPA)	HPA →
Cervix	Reported tissue cells	Medium	Protein (HPA)	HPA →
Cerebellum	Reported tissue cells	Medium	Protein (HPA)	HPA →
Cerebral cortex	Reported tissue cells	Medium	Protein (HPA)	HPA →

Undetected expression · recommended negative controls

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

Section 1

What Is the Expected GRB14 Western Blot Band Size?

Use the product-observed 61 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
61 kDa	Matches the authoritative product WB observation.
61 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 GRB14 appearancePlan around 61 kDa and keep the calculated mass as supporting context.

How each factor affects band size
Catalog-observed band	61 kDa; use this as the primary experimental expectation.
Calculated mass	61 kDa from UniProt Q14449; retain as context.
Gel selection	12-15%; shared with the recommended protocol and poster.
Specificity check	Compare the lead HPA positive and negative controls with A04177-2.

Why is my band missing or off?
Situation	Likely cause	Next action
61 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 Q14449 Human Protein Atlas product datasheet (A04177-2)

Experiment evidence

Western blot images showing GRB14 band

A04177-2 · GRB14 — observed 61 kDa WB validation

A04177-2 · GRB14Observed 61 kDa · product-specific WB image

← swipe / scroll for more images →

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

Section 2

Real Curated GRB14 Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysate	Caudate
Gel %	12-15%
Load	20-30 µg total protein per lane
Transfer	Semi-dry, standard transfer
Membrane	0.45 µm PVDF
Blocking	5% non-fat milk or 5% BSA in TBST
Primary	A04177-2 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 4

Advanced GRB14 Western Blot Tips

Deeper troubleshooting and optimisation questions for GRB14, answered from its protein features.

Which band should guide the blot?

Use 61 kDa, the observation attached to the authoritative A04177-2 WB record.

How should calculated mass be interpreted?

Treat the UniProt calculated mass as context; it does not replace the catalog-observed 61 kDa expectation.

Which positive control should I start with?

Start with Caudate, the lead HPA protein-expression candidate.

Which negative control is defensible?

Use Appendix as an orthogonal HPA Not detected candidate.

Which gel should I use?

Use 12-15% consistently across the quick facts, protocol table, and poster.

What transfer method to use for GRB14 Western blot?

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

How should A04177-2 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: PMC12627322, PMC2763493, PMC3878334.

Boster reagents

GRB14 Western Blot Reagents

Human-reactive GRB14 Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for GRB14 using A04177-2; observed band 61 kDa

Anti-GRB14 Antibody Picoband®

Cat # A04177-2

Reactivity: Human

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.