WDR5 / WD repeat-containing protein 5 · Western blot design guide

Design a Western Blot for WDR5

Real validated WDR5 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-WDR5 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 WDR5: expected band ~36.6 kDa, hero antibody A01910-3, catalog values and labelled standard workflow; separate PMC comparisons on the guide — Printable **WDR5** Western blot protocol sheet — expected band ~36.6 kDa, antibody A01910-3, controls and PMC citations. Open the full WDR5 WB guide →

Navigate to: Protocols Band Controls Antibodies

WDR5 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	~36.6 kDa
	Observed band	~37 kDa
	Gel	5–20% (catalog A01910-3)
	Negative control ⓘ	Suggested KO / knockdown lysate

Important caveats

Reasons your observed band may differ from the expected size.

	PTM	Acetylated + Ubl conjugation
	Caveat	Modification-state controls
	Gene-set association	MSigDB C7 membership
	Isoform	1 isoform(s)

Section 1

Real Curated WDR5 Western Blot Protocols

The A01910-3 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 293T, human Jurkat (catalog A01910-3)
Gel %	5–20% (catalog A01910-3)
Load	30 ug; reducing conditions (catalog A01910-3)
Transfer	a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A01910-3)
Membrane	nitrocellulose membrane (catalog A01910-3)
Blocking	5% non-fat milk/TBS for 1.5 hour at RT (catalog A01910-3)
Primary antibody	A01910-3 · 0.5 μg/mL (catalog A01910-3)
Primary incubation	overnight at 4°C (catalog A01910-3)
Secondary antibody	goat anti-rabbit IgG-HRP, 1:5000 (catalog A01910-3)
Secondary incubation	1.5 hour at RT (catalog A01910-3)
Wash	TBS-0.1%Tween 3 times with 5 minutes each (catalog A01910-3)
Detection	ECL (catalog A01910-3)

Section 2

What Is the Expected WDR5 Western Blot Band Size?

WDR5 is predicted at 36.6 kDa and observed at ~37 kDa; the small difference has no established cause in the supplied features.

What am I looking at on my blot?
Single band at ~37 kDa	Consistent with WDR5 in whole-cell lysate
Band near 36.6 kDa	Consistent with the UniProt predicted mass
~37 kDa band in a nuclear fraction	Consistent with WDR5 nuclear localization
No distinct acetylation doublet	The listed acetylation sites do not establish a resolvable shift

💡Expected WDR5 appearanceWDR5 has a predicted mass of 36.6 kDa and an observed band at ~37 kDa in whole-cell lysate; confirm identity with an appropriate antibody specificity control.

How each factor affects band size
UniProt predicted mass	Places the expected monomer band near 36.6 kDa; the observed band is ~37 kDa
334-residue sequence	Accounts for the predicted full-length mass without a listed cleavage event
N-acetylalanine at residue 2	No resolvable band shift is established
N6-acetyllysine at residue 112	No resolvable band shift is established

Why is my band missing or off?
Situation	Likely cause	Next action
No band in lysate	Nuclear WDR5 may be poorly recovered	Check nuclear extraction and run a positive-control whole-cell lysate
Band higher than expected	Identity of a band above ~37 kDa is unestablished	Check whether it decreases after WDR5 knockdown
Band lower than expected	Identity of a band below ~37 kDa is unestablished	Check sample integrity and WDR5 knockdown response
Multiple bands	A single listed isoform does not explain additional bands	Compare bands with a WDR5 knockdown control
Weak or no signal	Limited recovery of nuclear WDR5 or insufficient detection	Check nuclear protein recovery and a positive-control lysate

Source: UniProt P61964 Human Protein Atlas product datasheet (A01910-3)

Experiment evidence

Western blot images showing WDR5 band

A01910-3 · WDR5

M01910 · WDR5

A01910 · WDR5

← swipe / scroll for more images →

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

Sample controls for WDR5 Western blot

🧪For positive controls for WDR5 in Western blot, you can use no HPA-IHC candidate positive sample from the supplied 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 expression data are unavailable, so tissue controls cannot be validated from this evidence.

HPA tissue expression evidence for WDR5

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 WDR5 Western Blot Tips

Deeper troubleshooting and optimisation questions for WDR5, answered from its protein features.

How should WDR5 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 WDR5 isoforms explain additional bands?

Isoforms · Only one isoform is listed, with no alternative sequence. The supplied isoform information therefore does not explain additional bands; check their specificity before assigning them to WDR5.

Which WDR5 acetylation sites are annotated?

PTM · UniProt lists N-acetylalanine at position 2 and N6-acetyllysine at position 112. These are UniProt coordinates; check the numbering convention before comparing them with antibody or paper descriptions.

No visible shift follows from the annotations alone. Compare any shifted signal with the expected band near 37 kDa and confirm its identity before attributing it to acetylation.

Does this guide establish induction of WDR5?

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.

How should transfer be checked for WDR5?

Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.

How should blocking be optimized?

Blocking · Standard workflow guidance: follow the A01910-3 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 WDR5 be quantified?

Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.

Why is WDR5 observed near 37 kDa?

Interpretation · WDR5 has a predicted mass of 36.6 kDa, close to the reported apparent band near 37 kDa. The supplied features do not establish a modification-driven mass difference.

WDR5 is annotated in the nucleus. Include nuclear material when preparing samples, and keep sample preparation consistent across lanes being compared.

The listed single isoform and acetylation sites do not establish the identity of other bands. Treat bands away from approximately 37 kDa as unassigned until their WDR5 specificity is verified.

Boster reagents

WDR5 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 WDR5 using anti-WDR5 antibody (A01910-3). 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 30 ug of sample under reducing conditions. Lane 1: human 293T whole cell lysates, Lane 2: human Jurkat 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-WDR5 antigen affinity purified polyclonal antibody (Catalog # A01910-3) 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 WDR5 at approximately 37 kDa. The expected band size for WDR5 is at 37 kDa.

Anti-WDR5 Antibody Picoband®

Cat # A01910-3

View product →

Real WB data

Western blot analysis of WDR5 using anti-WDR5 antibody (M01910). 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 Jurkat whole cell lysates, Lane 2: human MOLT-4 whole cell lysates, Lane 3: 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-WDR5 antigen affinity purified monoclonal antibody (M01910) 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 WDR5 at approximately 37 kDa. The expected band size for WDR5 is at 37 kDa.

Anti-WDR5 Monoclonal Antibody

Cat # M01910

View product →

Real WB data Western blot analysis of WDR5 in 293 cell lysate with WDR5 antibody at 1 μg/ml in (A) the absence and (B) the presence of blocking peptide.

Anti-WD repeat-containing protein 5 WDR5 Antibody

Cat # A01910

View product →

Three the supplier anti-WDR5 antibodies have WB images. A01910-3 and M01910 show approximately 37 kDa bands in human cell lysates. A01910 shows a 293 cell lysate blot with and without blocking peptide. The supplied evidence does not establish performance across all reported species.

Which to pick: For human WB, choose by tested sample: A01910-3 shows 293T and Jurkat; M01910 shows Jurkat, MOLT-4 and MCF-7; A01910 shows 293 lysate with a blocking-peptide comparison. M01910 also lists mouse and rat reactivity, but its pictured WB uses human lysates.

Source: BosterBio WDR5 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.

References

UniProt Consortium. UniProt entry P61964.
Human Protein Atlas. WDR5 tissue expression.
PMC7573303 — target-verified WB comparison
PMC11623929 — target-verified WB comparison
PMC6011590 — target-verified WB comparison