SIRT1 · Western blot design guide

SIRT1 Western Blot Planning Guide

Plan a SIRT1 Western blot around the catalog-observed 81.7 kDa band, image-backed A00018-1 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 SIRT1 (SIRT1): expected band 81.7 kDa, antibody A00018-1, and PMC-cited SDS-PAGE protocol steps — SIRT1 Western blot protocol sheet — expected band 81.7 kDa, antibody A00018-1, controls and PMC citations. Open the full SIRT1 WB guide →

Navigate to: Protocols Band Controls Antibodies

SIRT1 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	81.7 kDa
	Observed band	Not reported — verify product WB image
	Gel	8-10%
	Positive control ⓘ	Testis
	Negative control ⓘ	Bronchus

Important caveats

Reasons your observed band may differ from the expected size.

ⓘ	Calculated mass	81.7 kDa
ⓘ	Localization	Nucleus, PML body / Cytoplasm
ⓘ	Processing / PTM	Record-dependent
ⓘ	Reactivity	Human

Section 1

Real Curated SIRT1 Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysate	Adrenal gland
Gel %	8-10%
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	A00018-1 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 2

What Is the Expected SIRT1 Western Blot Band Size?

Use the product-observed 81.7 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
81.7 kDa	Matches the authoritative product WB observation.
81.7 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 SIRT1 appearancePlan around 81.7 kDa and keep the calculated mass as supporting context.

How each factor affects band size
Catalog-observed band	81.7 kDa; use this as the primary experimental expectation.
Calculated mass	81.7 kDa from UniProt Q96EB6; retain as context.
Gel selection	8-10%; shared with the recommended protocol and poster.
Specificity check	Compare the lead HPA positive and negative controls with A00018-1.

Why is my band missing or off?
Situation	Likely cause	Next action
81.7 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 Q96EB6 Human Protein Atlas product datasheet (A00018-1)

Experiment evidence

Western blot images showing SIRT1 band

A00018-1 · SIRT1

← swipe / scroll for more images →

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

Sample controls for SIRT1 Western blot

🧪Use Testis as the first positive-control candidate and Bronchus as the HPA Not detected negative candidate.

Positive control: Testis (High)

Negative control: Bronchus (Not detected)

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

HPA tissue expression evidence for SIRT1

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.

Positive expression · recommended positive controls

Tissue	Cell type	Level	Evidence	Source
Testis	Reported tissue cells	High	Protein (HPA)	HPA →
Adrenal gland	Reported tissue cells	High	Protein (HPA)	HPA →
Lymph node	Reported tissue cells	High	Protein (HPA)	HPA →
Bone marrow	Reported tissue cells	Medium	Protein (HPA)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Bronchus	Reported tissue cells	Not detected	Protein (HPA)	HPA →
Cerebral cortex	Reported tissue cells	Not detected	Protein (HPA)	HPA →

Section 3

Advanced SIRT1 Western Blot Tips

Deeper troubleshooting and optimisation questions for SIRT1, answered from its protein features.

Which band should guide the blot?

Use 81.7 kDa, the observation attached to the authoritative A00018-1 WB record.

How should calculated mass be interpreted?

Treat the UniProt calculated mass as context; it does not replace the catalog-observed 81.7 kDa expectation.

Which positive control should I start with?

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

Which negative control is defensible?

Use Bronchus as an orthogonal HPA Not detected candidate.

Which gel should I use?

Use 8-10% consistently across the quick facts, protocol table, and poster.

What transfer method to use for SIRT1 Western blot?

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

How should A00018-1 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: PMC10206009, PMC12700963, PMC12782254.

Boster reagents

SIRT1 Western Blot Reagents

Human-reactive SIRT1 Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for SIRT1 using A00018-1; observed band 81.7 kDa

Anti-SIRT1 Antibody Picoband®

Cat # A00018-1

View product →

Real WB data Western blot validation image for SIRT1 using PB9159; observed band 81.7 kDa