SLC2A3 / Solute carrier family 2, facilitated glucose transporter member 3 · Western blot design guide

Design a Western Blot for SLC2A3

Real validated SLC2A3 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SLC2A3 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 SLC2A3: expected band ~53.9 kDa, hero antibody M03259, catalog values and labelled standard workflow; separate PMC comparisons on the guide — Printable **SLC2A3** Western blot protocol sheet — expected band ~53.9 kDa, antibody M03259, controls and PMC citations. Open the full SLC2A3 WB guide →

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SLC2A3 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	~53.9 kDa
	Observed band	~54 kDa
	Gel	5–20% (catalog M03259)
	Positive control ⓘ	Bone marrow (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	Glycosylated + Phosphorylated
	Caveat	Modification-state controls
	Gene-set association	MSigDB Hallmark membership
	Isoform	1 isoform(s)

Section 1

Real Curated SLC2A3 Western Blot Protocols

The M03259 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 HepG2, human 293T, human A431 (catalog M03259)
Gel %	5–20% (catalog M03259)
Load	30 ug; reducing conditions (catalog M03259)
Transfer	a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M03259)
Membrane	nitrocellulose membrane (catalog M03259)
Blocking	5% non-fat milk/TBS for 1.5 hour at RT (catalog M03259)
Primary antibody	M03259 · 1:500 (catalog M03259)
Primary incubation	overnight at 4°C (catalog M03259)
Secondary antibody	goat anti-rabbit IgG-HRP, 1:500 (catalog M03259)
Secondary incubation	1.5 hour at RT (catalog M03259)
Wash	TBS-0.1%Tween 3 times with 5 minutes each (catalog M03259)
Detection	ECL (catalog M03259)

Section 2

What Is the Expected SLC2A3 Western Blot Band Size?

SLC2A3 is predicted at 53.9 kDa and observed at ~54 kDa; the supplied evidence does not establish a cause for their small difference.

What am I looking at on my blot?
Band at ~54 kDa	Matches the empirical SLC2A3 band and its 53.9 kDa predicted mass.
Band above ~54 kDa	N-linked glycosylation at Asn43 could affect migration, but a visible shift is unproven.
Band near ~54 kDa without a visible shift	The Asn43 glycosylation site need not produce a resolvable change.
Weak or absent band in whole-cell lysate	Membrane localization may make SLC2A3 recovery sensitive to extraction.

💡Expected SLC2A3 appearanceSLC2A3 has a predicted mass of 53.9 kDa and an empirical band at ~54 kDa; confirm band identity with appropriate controls.

How each factor affects band size
Predicted SLC2A3 mass	53.9 kDa provides the reference for the empirical ~54 kDa band.
N-linked glycosylation site at Asn43	May affect apparent mass if occupied; no visible shift is established.
Occupied Asn43 glycosylation site	Could alter migration relative to protein without this modification; the magnitude is unknown.
Unoccupied Asn43 glycosylation site	Would lack any mass contribution from glycosylation at this site.

Why is my band missing or off?
Situation	Likely cause	Next action
No band in lysate	Membrane protein may be poorly recovered during extraction.	Check membrane protein extraction and include a positive control lysate.
Band higher than expected	Asn43 glycosylation could affect migration, but the cause is unestablished.	Compare with a validated positive control and assess glycosylation if needed.
Band lower than expected	Band identity or sample integrity is uncertain.	Check sample integrity and confirm identity with an independent antibody or knockdown.
Multiple bands	Additional bands are not explained by the single listed isoform.	Compare with a positive control and test which bands respond to knockdown.
Weak or no signal	Membrane protein recovery or SLC2A3 abundance may be low.	Check extraction, loading, and a positive control lysate.

Source: UniProt P11169 Human Protein Atlas product datasheet (M03259)

Experiment evidence

Western blot images showing SLC2A3 band

M03259 · SLC2A3

← swipe / scroll for more images →

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

Sample controls for SLC2A3 Western blot

🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SLC2A3 in Western blot, you can use bone marrow tissue, where HPA reports high expression.

Positive control: Bone marrow (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: As a multi-pass membrane protein, SLC2A3 may be easier to detect in membrane-enriched lysates.

HPA tissue expression evidence for SLC2A3

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
Bone marrow	hematopoietic cells	High	Protein (IHC)	HPA →
Spleen	cells in red pulp	High	Protein (IHC)	HPA →
Testis	elongated or late spermatids	High	Protein (IHC)	HPA →
Tonsil	non-germinal center cells	High	Protein (IHC)	HPA →
Appendix	lymphoid tissue	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 →
Bronchus	respiratory epithelial cells	Not detected	Protein (IHC)	HPA →
Caudate	glial cells	Not detected	Protein (IHC)	HPA →
Cerebellum	cells in granular layer	Not detected	Protein (IHC)	HPA →
Cervix	glandular cells	Not detected	Protein (IHC)	HPA →

Section 3

Advanced SLC2A3 Western Blot Tips

Deeper troubleshooting and optimisation questions for SLC2A3, answered from its protein features.

How should SLC2A3 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 alternative isoforms explain multiple SLC2A3 bands?

Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no isoform-based explanation for multiple bands. Check band identity before assigning additional bands to SLC2A3.

Could glycosylation affect SLC2A3 band migration?

PTM · UniProt lists one N-linked glycosylation site at Asn43. Compare untreated and deglycosylated samples if migration differs, but site annotation alone does not establish a visible shift.

UniProt lists phosphothreonine at 232 and 492, and phosphoserine at 475 and 485. These are UniProt coordinates; paper or antibody numbering may differ. The site annotations alone do not predict separate bands.

Does this guide establish induction of SLC2A3?

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 SLC2A3 Western blot?

Transfer · SLC2A3 is a multi-pass cell-membrane protein with a predicted mass of 53.9 kDa. Use a transfer setup that retains proteins around 54 kDa, then check the membrane and post-transfer gel for transfer. The supplied features do not specify one preferred method.

How should blocking be optimized?

Blocking · Standard workflow guidance: follow the M03259 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 SLC2A3 bands be quantified?

Quantitation · Quantify the same band near 54 kDa across samples using consistent sample preparation and loading normalization. SLC2A3 is a cell-membrane protein, so keep the analyzed sample fraction consistent. Treat other bands separately until their identity is established.

Why is the SLC2A3 band near 54 kDa?

Interpretation · The observed band near 54 kDa closely matches the predicted 53.9 kDa mass. The listed modifications do not establish a visible shift or explain any difference from the calculated mass.

Start with the expected band near 54 kDa. SLC2A3 has an annotated N-linked site at Asn43 and phosphorylation sites at UniProt positions 232, 475, 485 and 492, but these features alone do not identify an unexpected band or establish a visible shift. Verify band identity before attributing it to a modification.

Boster reagents

SLC2A3 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 GLUT3 using anti-GLUT3 antibody (M03259). 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 HepG2 whole cell lysates, Lane 2: human 293T whole cell lysates, Lane 3: human A431 whole cell lysates, Lane 4: human SH-SY5Y whole cell lysates, Lane 5: rat kidney tissue lysates, Lane 6: rat brain tissue lysates, Lane 7: mouse kidney tissue lysates, Lane 8: mouse brain 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-GLUT3 antigen affinity purified monoclonal antibody (Catalog # M03259) 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 1:500 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 GLUT3 at approximately 54 kDa. The expected band size for GLUT3 is at 54 kDa.

Anti-GLUT3 SLC2A3 Rabbit Monoclonal Antibody

Cat # M03259

View product →

M03259 is a rabbit monoclonal anti-SLC2A3 (GLUT3) antibody with a Western blot image showing an approximately 54 kDa band in the listed human cell, rat tissue, and mouse tissue lysates. This documents those tested samples and conditions only.

Which to pick: M03259 is the only listed option. Its stated reactivity is human, mouse, and rat; the WB image uses 30 µg lysate per lane and a 1:500 primary dilution. Match your sample to the reported contexts when planning a blot.

Source: BosterBio SLC2A3 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.

References

UniProt Consortium. UniProt entry P11169.
Human Protein Atlas. SLC2A3 tissue expression.
PMC3853007 — target-verified WB comparison
PMC9603975 — target-verified WB comparison
PMC11020985 — target-verified WB comparison