PCF11 / Pre-mRNA cleavage complex 2 protein Pcf11 · Western blot design guide

Design a Western Blot for PCF11

Source-linked PCF11 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PCF11 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 PCF11: expected band ~173.1 kDa, hero antibody A05372-1, catalog values and labelled standard workflow; separate PMC comparisons on the guide — Printable **PCF11** Western blot protocol sheet — expected band ~173.1 kDa, antibody A05372-1, controls and PMC citations. Open the full PCF11 WB guide →

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PCF11 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	~173.1 kDa
	Observed band	~200 kDa
	Gel	5–20% (catalog A05372-1)
	Negative control ⓘ	Suggested KO / knockdown lysate

Important caveats

Reasons your observed band may differ from the expected size.

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

Section 1

Source-Linked PCF11 Western Blot Protocol Options

The A05372-1 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 K562, human MCF-7, rat testis, rat C6, mouse testis (catalog A05372-1)
Gel %	5–20% (catalog A05372-1)
Load	30 ug; reducing conditions (catalog A05372-1)
Transfer	a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A05372-1)
Membrane	nitrocellulose membrane (catalog A05372-1)
Blocking	5% non-fat milk/TBS for 1.5 hour at RT (catalog A05372-1)
Primary antibody	A05372-1 · 0.5 μg/mL (catalog A05372-1)
Primary incubation	overnight at 4°C (catalog A05372-1)
Secondary antibody	goat anti-rabbit IgG-HRP, 1:5000 (catalog A05372-1)
Secondary incubation	1.5 hour at RT (catalog A05372-1)
Wash	TBS-0.1%Tween 3 times with 5 minutes each (catalog A05372-1)
Detection	ECL (catalog A05372-1)

Section 2

What Is the Expected PCF11 Western Blot Band Size?

PCF11 is predicted at 173.1 kDa and observed near 200 kDa on Western blots; the cause of this difference is not established.

What am I looking at on my blot?
Band near 200 kDa	Empirical PCF11 band in reducing whole-cell lysates
Band near 173 kDa	Near the predicted mass of full-length PCF11; identity requires confirmation
Closely spaced bands near the main band	Could reflect phosphorylation at annotated sites; distinct migration is unproven
Little signal in a cytoplasmic fraction	PCF11 is annotated as nuclear

💡Expected PCF11 appearancePCF11 has a predicted mass of 173.1 kDa, while antibody QC detected a band near 200 kDa in reducing whole-cell lysates; the cause of the difference is unestablished, so confirm identity with appropriate controls.

How each factor affects band size
Predicted full-length mass	173.1 kDa from the UniProt sequence
Phosphorylation at Ser120	May affect mobility; no visible shift is established
Phosphorylation at Thr121	May affect mobility; no visible shift is established
N-terminal acetylation at Ser2	Adds a small modification; no visible shift is established

Why is my band missing or off?
Situation	Likely cause	Next action
No band in lysate	Nuclear PCF11 may be poorly recovered during extraction	Check the nuclear fraction and extraction quality
Band higher than expected	The reported PCF11 band is near 200 kDa despite a 173.1 kDa predicted mass; the cause is unestablished	Compare with the reported band and confirm identity by PCF11 depletion
Band lower than expected	A lower band is not explained by an annotated signal peptide or propeptide	Check PCF11 depletion and sample integrity
Multiple bands	Annotated phosphorylation could affect mobility, but distinct bands are unproven	Compare phosphatase-treated samples and confirm bands by PCF11 depletion
Weak or no signal	Nuclear PCF11 may be underrepresented in the tested fraction	Test a nuclear-enriched sample with a positive control
Fragments below expected size	Fragment identity is not established by the supplied features	Check sample integrity and whether the fragments diminish after PCF11 depletion

Source: UniProt O94913 Human Protein Atlas product datasheet (A05372-1)

Experiment evidence

Western blot images showing PCF11 band

A05372-1 · PCF11

← swipe / scroll for more images →

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

Sample controls for PCF11 Western blot

🧪For positive controls for PCF11 in Western blot, you can use no HPA-supported positive sample because none is provided.

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 expression data, so tissue controls cannot be selected from this evidence.

HPA tissue expression evidence for PCF11

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 PCF11 Western Blot Tips

Deeper troubleshooting and optimisation questions for PCF11, answered from its protein features.

How should PCF11 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 PCF11 isoforms explain multiple bands?

Isoforms · The supplied UniProt record lists one isoform and no alternative sequence. It does not support assigning additional bands to annotated isoforms. Check their response to PCF11 depletion before identifying them as PCF11.

Which PCF11 phosphorylation sites matter when comparing bands?

PTM · UniProt assigns WNK1 phosphorylation to Ser120 and Thr121. It also lists phosphorylation at Ser169, Ser182, Thr459, Ser489, Ser494, Ser509, Ser511, Ser645, Ser705, Ser728, Ser777, Thr785, Ser794, Ser851, Ser1161, and Thr1530. These are UniProt coordinates; antibody or paper numbering may differ. Site annotations alone do not establish a detectable band shift.

UniProt lists N-acetylserine at position 2 and asymmetric dimethylarginine at positions 805, 820, 833, 929, 942, 955, 981, 994, 1007, 1093, and 1104. These positions use UniProt numbering. The annotations do not show that any modification changes the observed band position.

Does this guide establish induction of PCF11?

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 PCF11 Western blot?

Transfer · PCF11 is predicted at 173.1 kDa and has an observed band near 200 kDa, so verify that your transfer conditions recover proteins in that size range. Inspect the membrane and residual gel around 200 kDa, then adjust transfer time or conditions if substantial protein remains in the gel.

How should blocking be optimized?

Blocking · Standard workflow guidance: follow the A05372-1 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 the PCF11 band?

Quantitation · Quantify a consistently defined band near the supplied apparent size of 200 kDa, using exposures within the linear signal range and a suitable loading reference. PCF11 is annotated as nuclear, so keep sample preparation consistent when comparing nuclear fractions. Total-band intensity alone cannot measure occupancy of its listed modification sites.

Why might PCF11 appear near 200 kDa instead of 173.1 kDa?

Interpretation · The supplied apparent band is about 200 kDa, while the predicted mass is 173.1 kDa. UniProt lists phosphorylation, methylation, and acetylation, but their presence alone does not explain the difference or establish a visible shift. Confirm the band with an independent PCF11 antibody or PCF11 depletion.

The supplied record lists one isoform, no signal peptide or propeptide, and an observed band near 200 kDa. It provides no basis to label other bands as alternative isoforms or processed PCF11. Compare them with the expected band and test whether they respond to PCF11 depletion or an independent antibody.

Boster reagents

PCF11 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 PCF11 using anti-PCF11 antibody (A05372-1). 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 30 ug of sample under reducing conditions. Lane 1: human K562 whole cell lysates, Lane 2: human MCF-7 whole cell lysates, Lane 3: rat testis tissue lysates, Lane 4: rat C6 whole cell lysates, Lane 5: mouse testis tissue 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-PCF11 antigen affinity purified polyclonal antibody (Catalog # A05372-1) 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 PCF11 at approximately 200 kDa. The expected band size for PCF11 is at 173 kDa.

Anti-PCF11 Antibody Picoband®

Cat # A05372-1

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The catalog reports one anti-PCF11 antibody, A05372-1, for Western blotting. Its image shows human K562 and MCF-7, rat testis and C6, and mouse testis lysates. The reported band is approximately 200 kDa versus an expected 173 kDa; no independent validation is supplied.

Which to pick: A05372-1 is the only listed option. Its stated reactivity is human, mouse, and rat, and it has a Western blot image using the specific samples above. Consider the reported 200 kDa band when interpreting results.

Source: BosterBio PCF11 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.

References

UniProt Consortium. UniProt entry O94913.
Human Protein Atlas. PCF11 tissue expression.
PMC8934653 — target-verified WB comparison
PMC6458999 — target-verified WB comparison