SATB2 / DNA-binding protein SATB2 · Western blot design guide

Design a Western Blot for SATB2

Real validated SATB2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SATB2 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 SATB2: expected band ~82.6 kDa, hero antibody A02588, catalog values and labelled standard workflow; separate PMC comparisons on the guide — Printable **SATB2** Western blot protocol sheet — expected band ~82.6 kDa, antibody A02588, controls and PMC citations. Open the full SATB2 WB guide →

Navigate to: Protocols Band Controls Antibodies

SATB2 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	~82.6 kDa
	Gel	8–10% (standard starting point)
	Positive control ⓘ	Appendix (IHC candidate; verify WB) +4 more
	Negative control ⓘ	Adipose tissue (IHC candidate; verify WB)

Important caveats

Reasons your observed band may differ from the expected size.

	PTM	Phosphorylated + Ubl conjugation
	Caveat	Phosphorylation-state controls
	Gene-set association	MSigDB C7 membership
	Isoform	2 isoform(s)

Section 1

Real Curated SATB2 Western Blot Protocols

The A02588 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	mouse brain tissue lysate (catalog A02588)
Gel %	8–10% (standard starting point)
Load	20–30 µg total protein per lane; optimize for abundance (standard starting point)
Transfer	Standard semi-dry transfer; verify efficiency (standard starting point)
Membrane	0.45 µm PVDF (standard starting point)
Blocking	5% milk or 5% BSA in TBST (standard starting point)
Primary antibody	A02588 · (A) 2 and (B) 4 μg/mL (catalog A02588)
Primary incubation	Overnight at 4 °C (standard starting point)
Secondary antibody	Species-matched HRP conjugate at validated dilution (standard starting point)
Secondary incubation	1 h at room temperature (standard starting point)
Wash	3 × 5 min in TBST (standard starting point)
Detection	ECL; bracket exposures to avoid saturation (standard starting point)

Section 2

What Is the Expected SATB2 Western Blot Band Size?

SATB2 is predicted at 82.6 kDa; isoforms and phosphorylation could affect migration, but no empirical band size or visible effect is established.

What am I looking at on my blot?
Band near 82.6 kDa	Consistent with the predicted SATB2 mass; identity requires controls
Several bands at different positions	Isoforms 1 and 2 are possible contributors, but their migration is unknown
Doublet near the expected position	Possible isoform or phosphorylation effects; neither is demonstrated here
Little or no band in soluble lysate	Nuclear matrix SATB2 may be poorly extracted

💡Expected SATB2 appearanceSATB2 has a predicted mass of 82.6 kDa, but no empirical band size is supplied; use an appropriate positive control and SATB2 depletion to assess band identity.

How each factor affects band size
Predicted mass of 82.6 kDa	Provides a sequence-based reference for band position
Isoform 1	Its individual apparent size is not supplied
Isoform 2	Its size relative to isoform 1 is not supplied
Alternative splicing	Could yield different band positions, but distinct migration is unconfirmed

Why is my band missing or off?
Situation	Likely cause	Next action
No band in lysate	Nuclear matrix SATB2 may resist extraction	Check a nuclear matrix fraction and a positive control
Band higher than expected	The supplied features do not establish the cause of slower migration	Compare with a positive control and test band identity by SATB2 depletion
Band lower than expected	An isoform is possible, but its mass is unknown	Check band identity by SATB2 depletion
Multiple bands	Isoforms 1 and 2 are possible contributors	Use SATB2 depletion to identify specific bands
Weak or no signal	Nuclear matrix SATB2 may be underrepresented in the extract	Assess nuclear matrix extraction and run a positive control

Source: UniProt Q9UPW6 Human Protein Atlas product datasheet (A02588)

Experiment evidence

Western blot images showing SATB2 band

A02588 · SATB2

M02588 · SATB2

A02588-2 · SATB2

M02588-2 · SATB2

← swipe / scroll for more images →

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

Sample controls for SATB2 Western blot

🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SATB2 in Western blot, you can use appendix tissue, which HPA scores as high expression.

Positive control: Appendix (IHC candidate; verify WB)

Negative control: Adipose tissue (IHC candidate; verify WB)

Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.

⚠️Feasibility: SATB2 is nuclear matrix localized, so nuclear-enriched lysate may improve detection.

HPA tissue expression evidence for SATB2

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
Appendix	endocrine cells	High	Protein (IHC)	HPA →
Cerebral cortex	neuronal cells	High	Protein (IHC)	HPA →
Colon	endocrine cells	High	Protein (IHC)	HPA →
Rectum	endocrine cells	High	Protein (IHC)	HPA →
Small intestine	endocrine cells	Medium	Protein (IHC)	HPA →

Lower expression tissues · IHC evidence, not confirmed WB-negative controls

Tissue	Cell type	Level	Evidence	Source
Adipose tissue	adipocytes	Not detected	Protein (IHC)	HPA →
Adrenal gland	glandular cells	Not detected	Protein (IHC)	HPA →
Bone marrow	hematopoietic cells	Not detected	Protein (IHC)	HPA →
Breast	adipocytes	Not detected	Protein (IHC)	HPA →
Bronchus	respiratory epithelial cells	Not detected	Protein (IHC)	HPA →

Section 3

Advanced SATB2 Western Blot Tips

Deeper troubleshooting and optimisation questions for SATB2, answered from its protein features.

How should SATB2 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 SATB2 isoforms produce different bands?

Isoforms · UniProt lists isoforms 1 and 2. Isoform 2 lacks residues 116–233 of the canonical sequence, so it may run lower. Check whether the antibody recognizes a region retained in isoform 2 before interpreting a second band.

Which SATB2 phosphorylation sites matter when interpreting bands?

PTM · UniProt lists phosphoserine at positions 20, 39, 454 and 594, and phosphothreonine at position 467. These are UniProt canonical-sequence coordinates; antibody or paper numbering may differ. Their presence alone does not establish a visible band shift.

Does this guide establish induction of SATB2?

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 SATB2?

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 A02588 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 SATB2 across samples?

Quantitation · Keep extraction consistent because SATB2 is annotated in the nuclear matrix. If multiple bands appear, define which band or bands you quantify and apply that choice consistently; the listed isoforms and phosphorylation sites do not establish the identity of any observed band.

What molecular weight should I expect for SATB2?

Interpretation · The predicted mass is 82.6 kDa for the 733-residue canonical sequence. No observed Western blot band position is supplied, so use 82.6 kDa as a reference rather than an established apparent mass.

SATB2 is annotated in the nuclear matrix. Include a nuclear matrix-containing fraction when preparing samples, and use the same extraction approach across samples to make band intensities comparable.

First compare it with the 82.6 kDa predicted canonical mass and consider isoform 2, which lacks residues 116–233. Check the antibody’s recognized region. The five listed phosphorylation sites do not, by themselves, explain an observed mass difference or prove that a band is SATB2.

Boster reagents

SATB2 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 SATB2 in mouse brain tissue lysate with SATB2 antibody at (A) 2 and (B) 4 μg/mL.

Anti-DNA-binding protein SATB2 SATB2 Antibody

Cat # A02588

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Real WB data

Western blot analysis of SATB2 using anti-SATB2 antibody (M02588). 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 THP-1 whole cell lysates, Lane 2: human K562 whole cell lysates, Lane 3: human HEL 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-SATB2 antigen affinity purified monoclonal antibody (Catalog # M02588) 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 1:1000 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 SATB2 at approximately 83 kDa. The expected band size for SATB2 is at 83 kDa.

Anti-SATB2 Rabbit Monoclonal Antibody

Cat # M02588

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Real WB data

Western blot analysis of SATB2 using anti-SATB2 antibody (A02588-2). 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 HEL 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-SATB2 antigen affinity purified polyclonal antibody (Catalog # A02588-2) 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 SATB2 at approximately 90 kDa. The expected band size for SATB2 is at 83 kDa.

Anti-SATB2 Antibody Picoband®

Cat # A02588-2

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Real WB data

Western blot analysis of SATB2 using anti-SATB2 antibody (M02588-2). 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 HT-1080 whole cell lysates, Lane 2: human HEL whole cell lysates, Lane 3: human THP-1 whole cell lysates, Lane 4: human K562 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-SATB2 antigen affinity purified monoclonal antibody (M02588-2) 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 SATB2 at approximately 90 kDa. The expected band size for SATB2 is at 90 kDa.

Anti-SATB2 Rabbit Monoclonal Antibody

Cat # M02588-2

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All four listed anti-SATB2 antibodies have Western blot images: A02588 in mouse brain, and M02588, A02588-2, and M02588-2 in human cell lysates. Reported bands are approximately 83 or 90 kDa. These images document the named samples only.

Which to pick: For mouse brain, consider A02588. For human lysates, M02588 reports an 83-kDa band in three cell lines, M02588-2 reports 90 kDa in four, and A02588-2 reports 90 kDa in HEL. A02588 and M02588 list rat reactivity, without rat blot examples.

Source: BosterBio SATB2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.

References

UniProt Consortium. UniProt entry Q9UPW6.
Human Protein Atlas. SATB2 tissue expression.
PMC6834402 — target-verified WB comparison
PMC3190908 — target-verified WB comparison
PMC9924951 — target-verified WB comparison