This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
GLUD1 / Glutamate dehydrogenase 1, mitochondrial · Western blot design guide
Design a Western Blot for GLUD1
Real validated GLUD1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-GLUD1 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
GLUD1 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 | ~61.4 kDa | |
| Observed band | ~50 kDa | |
| Gel | 10% (catalog M01866) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Band identity controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated GLUD1 Western Blot Protocols
The M01866 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human HepG2, human A431, rat liver (catalog M01866) |
| Gel % | 10% (catalog M01866) |
| Load | 30 ug; reducing conditions (catalog M01866) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M01866) |
| Membrane | nitrocellulose membrane (catalog M01866) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M01866) |
| Primary antibody | M01866 · 1:500 (catalog M01866) |
| Primary incubation | overnight at 4°C (catalog M01866) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog M01866) |
| Secondary incubation | 1.5 hour at RT (catalog M01866) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M01866) |
| Detection | ECL (catalog M01866) |
Section 2
What Is the Expected GLUD1 Western Blot Band Size?
GLUD1 is predicted at 61.4 kDa, while antibody QC observes ~50 kDa; the cause of the difference is not established.
| Band near 50 kDa | Empirical GLUD1 band reported in antibody QC; confirm identity with controls. |
| Band near 61.4 kDa | Near the predicted sequence mass; identity requires confirmation. |
| Several bands at different positions | Isoforms 1, 2, and 3 are annotated, but distinct migration is unproven. |
| Faint band in a mitochondria-poor sample | GLUD1 localizes mostly to mitochondria. |
💡Expected GLUD1 appearanceUniProt predicts 61.4 kDa, while antibody QC reports a ~50 kDa GLUD1 band; the cause of the difference is unestablished, so confirm band identity with appropriate controls.
| UniProt predicted mass | 61.4 kDa is the sequence-based benchmark; antibody QC reports ~50 kDa, with the difference unexplained. |
| Splice isoform 1 | Its individual apparent size is not supplied. |
| Splice isoform 2 | Its size relative to isoform 1 is not supplied. |
| Splice isoform 3 | Its size relative to the other isoforms is not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Poor recovery of mitochondria, where most GLUD1 localizes. | Check mitochondrial recovery and compare with a mitochondria-enriched fraction. |
| Band higher than expected | The band's identity or isoform contribution is uncertain. | Compare with a GLUD1-depleted control and an independent antibody. |
| Band lower than expected | The reported ~50 kDa band lies below the 61.4 kDa prediction; its migration difference is unexplained. | Confirm identity with GLUD1 depletion and an independent antibody. |
| Multiple bands | Three splice isoforms are annotated, but their migration is unknown. | Test which bands respond to GLUD1 depletion. |
| Weak or no signal | The sample may contain little mitochondrial GLUD1. | Check sample loading and mitochondrial recovery. |
Experiment evidence
Western blot images showing GLUD1 band
M01866 · GLUD1
A01866-3 · GLUD1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for GLUD1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for GLUD1 in Western blot, you can use adrenal gland tissue, which HPA scores High.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA lists no Not-detected tissue, so use siRNA knockdown or a KO line for a clear negative control.
HPA tissue expression evidence for GLUD1
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | glial cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bone marrow | hematopoietic cells | Low | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Low | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Colon | endothelial cells | Medium | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced GLUD1 Western Blot Tips
Deeper troubleshooting and optimisation questions for GLUD1, answered from its protein features.
How should GLUD1 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 GLUD1 isoforms affect band interpretation?
Isoforms · UniProt lists three isoforms. Relative to the canonical sequence, isoform 2 lacks residues 1–167; isoform 3 replaces residues 1–16 and lacks residues 17–149. These changes could affect apparent size and antibody recognition. Check whether the antibody epitope is retained before assigning a band to an isoform.
Which GLUD1 modifications matter when comparing samples?
PTM · UniProt lists phosphorylation at Ser79, Ser128, Ser227, Ser384, Thr410, Tyr135, and Tyr512, plus numerous lysine modifications. For example, Lys457, Lys503, and Lys527 each have alternate acetylation, malonylation, and succinylation annotations. These are canonical UniProt coordinates; antibody or paper numbering may differ. Their presence does not establish a visible band shift.
The supplied UniProt features list no glycosylation sites. Do not assign a band difference to glycosylation from this record. Investigate band identity and the listed isoform differences before interpreting an unexpected size.
The supplied UniProt features list no glycosylation sites. Do not assign a band difference to glycosylation from this record. Investigate band identity and the listed isoform differences before interpreting an unexpected size.
Does this guide establish induction of GLUD1?
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 GLUD1?
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 M01866 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 GLUD1 bands be quantified across samples?
Quantitation · Quantify the same validated band in each sample and keep the sampled fraction consistent. Because GLUD1 has three isoforms and is predominantly mitochondrial, a change in band intensity could reflect a different isoform contribution or mitochondrial enrichment rather than a change in total GLUD1.
Why might GLUD1 appear near 50 kDa instead of 61.4 kDa?
Interpretation · The observed band is approximately 50 kDa, while the predicted mass is 61.4 kDa. GLUD1 has alternative isoforms and a transit peptide annotation, but these features alone do not establish which form produces the band or explain the difference. Confirm band identity with an independent GLUD1 antibody or another identification method.
GLUD1 is reported mostly in mitochondria, with a small amount in the endoplasmic reticulum. Compare the same fraction across samples and account for differences in mitochondrial enrichment when interpreting band intensity.
Check whether their sizes and antibody epitopes are compatible with the listed isoforms, particularly the N-terminal deletions in isoforms 2 and 3. The homohexamer annotation and modification sites do not, by themselves, identify an extra band. Validate its identity before including it in quantitation.
GLUD1 is reported mostly in mitochondria, with a small amount in the endoplasmic reticulum. Compare the same fraction across samples and account for differences in mitochondrial enrichment when interpreting band intensity.
Check whether their sizes and antibody epitopes are compatible with the listed isoforms, particularly the N-terminal deletions in isoforms 2 and 3. The homohexamer annotation and modification sites do not, by themselves, identify an extra band. Validate its identity before including it in quantitation.
Boster reagents
GLUD1 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
Anti-GLUD1 Rabbit Monoclonal Antibody
Cat # M01866
Real WB data
Anti-GLUD1 Antibody Picoband®
Cat # A01866-3
Both listed anti-GLUD1 antibodies have Western blot images using human, rat, and mouse lysates. M01866 shows a band near 50 kDa and A01866-3 near 55 kDa, versus the stated 61 kDa expectation. The supplied evidence does not establish band identity independently.
Which to pick: Both list human, mouse, and rat reactivity. Choose A01866-3 if its illustrated HeLa, PC-3, or C2C12 lysates match your experiment; M01866 also has illustrated HepG2, A431, rat liver, C6, and mouse liver lysates. Compare the reported band positions before choosing.
Source: BosterBio GLUD1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P00367.
- Human Protein Atlas. GLUD1 tissue expression.
- PMC10319899 — target-verified WB comparison
- PMC10786780 — target-verified WB comparison
- PMC11321297 — target-verified WB comparison