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- Table of Contents
GCLM / Glutamate--cysteine ligase regulatory subunit · Western blot design guide
Design a Western Blot for GCLM
Real validated GCLM Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-GCLM 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
GCLM 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 | ~30.7 kDa | |
| Observed band | Approximately 31 kDa | |
| Gel | 5–20% (catalog A02948-2) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Ovary (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated GCLM Western Blot Protocols
The A02948-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human 293T, human Jurkat, human Hela (catalog A02948-2) |
| Gel % | 5–20% (catalog A02948-2) |
| Load | 30 ug; reducing conditions (catalog A02948-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A02948-2) |
| Membrane | nitrocellulose membrane (catalog A02948-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A02948-2) |
| Primary antibody | A02948-2 · 0.5 μg/mL (catalog A02948-2) |
| Primary incubation | overnight at 4°C (catalog A02948-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A02948-2) |
| Secondary incubation | 1.5 hour at RT (catalog A02948-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A02948-2) |
| Detection | ECL (catalog A02948-2) |
Section 2
What Is the Expected GCLM Western Blot Band Size?
GCLM is predicted at 30.7 kDa and observed at approximately 31 kDa; the small difference has no established cause in the supplied evidence.
| Band at approximately 31 kDa | Matches the empirical GCLM band, close to its 30.7 kDa predicted mass. |
| One band near 31 kDa | Isoforms 1 and 2 need not resolve into separate bands. |
| Additional band at a different size | Could reflect an isoform; its migration and identity require confirmation. |
| No distinct shifted band | Lys263 acetylation has no demonstrated visible migration effect. |
💡Expected GCLM appearanceGCLM has a predicted mass of 30.7 kDa and an observed band at approximately 31 kDa in whole-cell lysates; confirm band identity with appropriate controls.
| UniProt predicted mass | 30.7 kDa, close to the observed approximately 31 kDa band. |
| Isoform 1 | Its individual mass and migration are not supplied. |
| Isoform 2 | Its individual mass and migration are not supplied. |
| Alternative splicing | May yield different sizes, but separate bands are not established. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | GCLM was detected in whole-cell lysates, so absence has no established feature-based explanation. | Check a positive lysate, loading, transfer, and antibody performance. |
| Band higher than expected | An isoform is possible, but its migration is unknown. | Compare with a positive control and confirm band identity. |
| Band lower than expected | An isoform is possible, but its migration is unknown. | Compare with a positive control and confirm band identity. |
| Multiple bands | Isoforms 1 and 2 exist, but distinct bands are not established. | Use band-identity controls before assigning bands to isoforms. |
| Weak or no signal | No supplied feature establishes why signal would be weak. | Check loading, transfer, and a positive lysate. |
| Fragments below expected size | No supplied cleavage feature identifies these bands. | Check sample integrity and confirm band identity. |
Experiment evidence
Western blot images showing GCLM band
A02948-2 · GCLM
M02948 · GCLM
M02948-2 · GCLM
M02948-2 · GCLM
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for GCLM Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for GCLM in Western blot, you can use adrenal gland tissue, which HPA rates as medium expression.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Ovary (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA lists ovary as not detected, but the adrenal gland signal is only medium, so the positive control may be modest.
HPA tissue expression evidence for GCLM
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 | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Breast | glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Vagina | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Low | Protein (IHC) | HPA → |
| Cervix | glandular cells | Low | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced GCLM Western Blot Tips
Deeper troubleshooting and optimisation questions for GCLM, answered from its protein features.
How should GCLM 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 GCLM isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. Isoform 2 lacks residues 43–64 of the canonical sequence, so consider isoform expression when interpreting a smaller band. The sequence difference alone does not establish where either isoform will migrate.
If the epitope lies within canonical residues 43–64, an antibody may miss isoform 2, which lacks that segment. Check the antibody’s mapped epitope before treating a single detected band as total GCLM.
If the epitope lies within canonical residues 43–64, an antibody may miss isoform 2, which lacks that segment. Check the antibody’s mapped epitope before treating a single detected band as total GCLM.
Does acetylation explain a shifted GCLM band?
PTM · UniProt lists N6-acetyllysine at canonical residue 263. This modification alone does not demonstrate a visible band shift or explain any observed-versus-calculated mass difference. If comparing isoform numbering, account for isoform 2’s missing residues 43–64.
Does this guide establish induction of GCLM?
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 GCLM?
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 A02948-2 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 GCLM bands be quantified?
Quantitation · Decide whether the measurement should include both isoforms. An antibody recognizing a region shared by isoforms 1 and 2 is more suitable for that purpose than one targeting residues 43–64, which isoform 2 lacks. Record which bands are included in the measurement.
Why does GCLM appear near 31 kDa?
Interpretation · The supplied predicted mass is 30.7 kDa, and the observed band is approximately 31 kDa. These values are consistent; the listed features do not establish a band shift.
Consider isoform 2 if a smaller band appears: it lacks canonical residues 43–64. GCLM also forms a heterodimer with a catalytic heavy chain, but the supplied features do not assign an unexpected band to that complex. Verify band identity before interpreting it as GCLM.
Consider isoform 2 if a smaller band appears: it lacks canonical residues 43–64. GCLM also forms a heterodimer with a catalytic heavy chain, but the supplied features do not assign an unexpected band to that complex. Verify band identity before interpreting it as GCLM.
Boster reagents
GCLM 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-GCLM Antibody Picoband®
Cat # A02948-2
Real WB data
Anti-GCLM Monoclonal Antibody
Cat # M02948
Real WB data
Anti-GCLM Rabbit Monoclonal Antibody
Cat # M02948-2
Three the supplier anti-GCLM antibodies list human, mouse, and rat reactivity. A02948-2 has a WB image with human, rat, and mouse lysates and a reported 31 kDa band; M02948 shows K562 lysate. M02948-2 shows IP followed by WB, with limited context.
Which to pick: Choose A02948-2 for the most detailed direct WB example, including human 293T, Jurkat, HeLa, rat L6, and mouse NIH/3T3 lysates. M02948 has a K562 WB example. M02948-2 documents IP followed by WB; its image does not establish direct lysate WB performance.
Source: BosterBio GCLM gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P48507.
- Human Protein Atlas. GCLM tissue expression.
- PMC6185788 — target-verified WB comparison
- PMC2819613 — target-verified WB comparison
- PMC3625031 — target-verified WB comparison