KLF10 / Krueppel-like factor 10 · Western blot design guide

Design a Western Blot for KLF10

Real validated KLF10 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-KLF10 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 KLF10: expected band ~52.6 kDa, hero antibody A03419-2, catalog values and labelled standard workflow; separate PMC comparisons on the guide — Printable **KLF10** Western blot protocol sheet — expected band ~52.6 kDa, antibody A03419-2, controls and PMC citations. Open the full KLF10 WB guide →

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KLF10 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	~52.6 kDa
	Observed band	~55 kDa
	Gel	5–20% (catalog A03419-2)
	Negative control ⓘ	Suggested KO / knockdown lysate

Important caveats

Reasons your observed band may differ from the expected size.

	PTM	Phosphorylated + Ubl conjugation
	Caveat	Modification-state controls
	Gene-set association	MSigDB Hallmark membership
	Isoform	4 isoform(s)

Section 1

Real Curated KLF10 Western Blot Protocols

The A03419-2 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 Hela, rat brain, mouse skeletal muscle (catalog A03419-2)
Gel %	5–20% (catalog A03419-2)
Load	30 ug; reducing conditions (catalog A03419-2)
Transfer	a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A03419-2)
Membrane	nitrocellulose membrane (catalog A03419-2)
Blocking	5% non-fat milk/TBS for 1.5 hour at RT (catalog A03419-2)
Primary antibody	A03419-2 · 0.5 μg/mL (catalog A03419-2)
Primary incubation	overnight at 4°C (catalog A03419-2)
Secondary antibody	goat anti-rabbit IgG-HRP, 1:5000 (catalog A03419-2)
Secondary incubation	1.5 hour at RT (catalog A03419-2)
Wash	TBS-0.1%Tween 3 times with 5 minutes each (catalog A03419-2)
Detection	ECL (catalog A03419-2)

Section 2

What Is the Expected KLF10 Western Blot Band Size?

KLF10 is predicted at 52.6 kDa and observed around 55 kDa; the cause of the difference is not established.

What am I looking at on my blot?
Band around 55 kDa	Empirical KLF10 band reported in reducing lysates; confirm identity with controls
Band near 52.6 kDa	Compatible with KLF10's predicted mass; identity requires confirmation
Several bands at different masses	Could include splice isoforms 1–4; their migration is unreported
Close doublet	Could reflect phosphorylation at Ser184 or Ser249; visible separation is unestablished

💡Expected KLF10 appearanceKLF10 has a predicted mass of 52.6 kDa and an empirical band around 55 kDa in reducing lysates; the difference is unexplained, so confirm band identity with appropriate controls.

How each factor affects band size
Predicted KLF10 mass	52.6 kDa from sequence; the reported band is around 55 kDa, with no established cause for the difference
Splice isoform 1	May differ in size from other isoforms; its individual mass and migration are unreported
Splice isoform 2	May differ in size from other isoforms; its individual mass and migration are unreported
Splice isoform 3	May differ in size from other isoforms; its individual mass and migration are unreported
Splice isoform 4	May differ in size from other isoforms; its individual mass and migration are unreported

Why is my band missing or off?
Situation	Likely cause	Next action
No band in lysate	Nuclear KLF10 may be poorly recovered	Check a nuclear extract and a positive control
Band higher than expected	The reported band is around 55 kDa versus a predicted 52.6 kDa; the cause is unestablished	Compare with a validated positive control and confirm antibody specificity
Band lower than expected	An isoform is possible, but individual isoform masses are unknown	Confirm identity with an independent antibody or KLF10 depletion
Multiple bands	Splice isoforms 1–4 are possible, but distinct migration is unproven	Use KLF10 depletion or isoform-specific controls to assign bands
Weak or no signal	Nuclear KLF10 may be underrepresented in the sample	Check nuclear recovery and run a positive control

Source: UniProt Q13118 Human Protein Atlas product datasheet (A03419-2)

Experiment evidence

Western blot images showing KLF10 band

A03419-2 · KLF10

A03419 · KLF10

M03419 · KLF10

← swipe / scroll for more images →

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

Sample controls for KLF10 Western blot

🧪For positive controls for KLF10 in Western blot, you can use no HPA-supported tissue or cell line because none is 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: HPA data are unavailable, so a positive sample cannot be selected and a KLF10 knockdown or KO is the clearest negative control.

HPA tissue expression evidence for KLF10

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 KLF10 Western Blot Tips

Deeper troubleshooting and optimisation questions for KLF10, answered from its protein features.

How should KLF10 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.

How could KLF10 isoforms affect band patterns?

Isoforms · UniProt lists four isoforms. Isoform 2 changes the N terminus; isoforms 3 and 4 also have alternative sequence and large C-terminal deletions. Check which isoforms your antibody can recognize before assigning bands. The supplied features do not establish their apparent band sizes.

Possibly. Isoform 3 lacks canonical residues 103–480, and isoform 4 lacks 135–480. Check whether the antibody targets a region retained in each isoform before assigning a smaller band. Size alone cannot identify it.

Which KLF10 phosphorylation sites matter when interpreting bands?

PTM · UniProt lists phosphoserine at canonical positions 184 and 249. Those regions are missing from isoforms 3 and 4. Keep the canonical numbering explicit when comparing antibody or paper coordinates. Phosphorylation alone does not establish a visible shift.

Does this guide establish induction of KLF10?

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 KLF10?

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 A03419-2 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 KLF10 bands be quantified across samples?

Quantitation · Quantify the same clearly assigned band across samples. KLF10 has four listed isoforms, so first check whether your antibody can detect more than one. Report which band was measured; a change in one band does not establish a change in every isoform.

Why might KLF10 appear near 55 kDa instead of 52.6 kDa?

Interpretation · The observed band is about 55 kDa, versus a predicted mass of 52.6 kDa. Apparent migration can differ from calculated mass; the listed features do not establish the cause of this difference.

UniProt lists KLF10 in the nucleus. If using fractionated samples, examine the nuclear fraction and verify the fractionation before interpreting a weak or absent band.

Boster reagents

KLF10 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 KLF10 using anti-KLF10 antibody (A03419-2). 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 Hela whole cell lysates, Lane 2: rat brain tissue lysates, Lane 3: mouse skeletal muscle 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-KLF10 antigen affinity purified polyclonal antibody (Catalog # A03419-2) 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 KLF10 at approximately 55 kDa. The expected band size for KLF10 is at 53 kDa.

Anti-KLF10 Antibody Picoband®

Cat # A03419-2

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Real WB data Western blot (WB) analysis of KLF10/KLF11 polyclonal antibody at 1:500 dilution Lane1:HEK293T whole cell lysate Lane2:RAW264.7 whole cell lysate Lane3:PC12 whole cell lysate

Western blot (WB) analysis of KLF10/KLF11 polyclonal antibody at 1:500 dilution Lane1:HEK293T whole cell lysate Lane2:RAW264.7 whole cell lysate Lane3:PC12 whole cell lysate

Anti-KLF10/KLF11 Antibody

Cat # A03419

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Real WB data Western blot analysis of KLF10 expression in (1) HepG2 cell lysate; (2) NIH/3T3 cell lysate.

Anti-KLF10 Rabbit Monoclonal Antibody

Cat # M03419

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Three the supplier antibodies are listed for KLF10, each with a WB image. Their captions describe named cell or tissue lysates; these examples do not establish performance across all samples or independent target validation.

Which to pick: A03419-2 lists human reactivity and has a detailed WB caption covering HeLa, rat brain, and mouse muscle lysates. A03419 and M03419 list human, mouse, and rat reactivity; compare their pictured sample types with yours when choosing.

Source: BosterBio KLF10 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.

References

UniProt Consortium. UniProt entry Q13118.
Human Protein Atlas. KLF10 tissue expression.
PMC9988960 — target-verified WB comparison
PMC11895565 — target-verified WB comparison
PMC8141503 — target-verified WB comparison