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- Table of Contents
OGDH / 2-oxoglutarate dehydrogenase complex component E1 · Western blot design guide
Design a Western Blot for OGDH
Real validated OGDH Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-OGDH 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
OGDH 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 | ~115.9 kDa | |
| Observed band | ~110 kDa | |
| Gel | 5–20% (catalog A05301-1) | |
| 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 | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated OGDH Western Blot Protocols
The A05301-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human HepG2, human Daudi (catalog A05301-1) |
| Gel % | 5–20% (catalog A05301-1) |
| Load | 30 ug; reducing conditions (catalog A05301-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A05301-1) |
| Membrane | nitrocellulose membrane (catalog A05301-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A05301-1) |
| Primary antibody | A05301-1 · 0.5 μg/mL (catalog A05301-1) |
| Primary incubation | overnight at 4°C (catalog A05301-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A05301-1) |
| Secondary incubation | 1.5 hour at RT (catalog A05301-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A05301-1) |
| Detection | ECL (catalog A05301-1) |
Section 2
What Is the Expected OGDH Western Blot Band Size?
OGDH is predicted at 115.9 kDa and observed near 110 kDa in reducing lysates; the cause of this difference is not established.
| Band near 110 kDa | Empirical OGDH band in reducing whole-cell lysates; confirm identity with controls |
| Band near 116 kDa | Near the predicted 115.9 kDa monomer mass |
| Band near twice the monomer size | Could reflect a homodimer if it survives sample preparation |
| Several nearby bands | Could include splice isoforms 1, 2, and 3; their migration is unestablished |
💡Expected OGDH appearanceOGDH has a predicted mass of 115.9 kDa, while antibody QC detects a band near 110 kDa in reducing lysates; the difference is unexplained, so confirm identity with ordinary controls.
| Predicted OGDH mass | 115.9 kDa by sequence; antibody QC detects about 110 kDa |
| OGDH homodimer | Could migrate near twice the monomer size if preserved during sample preparation |
| Splice isoform 1 | Its individual mass and migration are not supplied |
| Splice isoform 2 | Its individual mass and migration are not supplied |
| Splice isoform 3 | Its individual mass and migration are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Mainly mitochondrial OGDH may be poorly recovered | Check mitochondrial protein recovery and a positive lysate control |
| Band higher than expected | A homodimer may persist during sample preparation | Compare fully denatured samples and confirm identity with OGDH depletion |
| Band lower than expected | The documented 110 kDa band is below the 115.9 kDa prediction; the cause is unknown | Compare with the antibody QC band and confirm by OGDH depletion |
| Multiple bands | Splice isoforms 1, 2, and 3 are annotated, but distinct migration is unproven | Use OGDH depletion or a second antibody to identify specific bands |
| Weak or no signal | Mainly mitochondrial localization may limit OGDH recovery | Check loading, mitochondrial recovery, and a positive control |
Experiment evidence
Western blot images showing OGDH band
A05301-1 · OGDH
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for OGDH Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for OGDH in Western blot, you can use adrenal gland tissue, which shows high HPA expression.
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 such as Ponceau alongside.
⚠️Feasibility: No HPA tissue is reported as not detected, so use siRNA knockdown or a KO line for a negative control.
HPA tissue expression evidence for OGDH
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 |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Cervix | glandular cells | Medium | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Medium | Protein (IHC) | HPA → |
| Hippocampus | glial cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced OGDH Western Blot Tips
Deeper troubleshooting and optimisation questions for OGDH, answered from its protein features.
How should OGDH 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 might OGDH isoforms affect band interpretation?
Isoforms · UniProt lists isoforms 1, 2, and 3. Isoform 2 replaces residues 139–172; isoform 3 replaces residues 404–427 and lacks residues 428–1023. Check whether the antibody epitope is retained in each isoform before assigning bands. Isoform 3 could yield a much smaller product if expressed and detected.
Possibly. Isoform 3 lacks canonical residues 428–1023 and has a replacement at 404–427. An antibody recognizing a retained region could detect it, but the feature record does not show whether it is expressed in the sample. Check the antibody epitope and compare the band with an isoform-specific reference.
Possibly. Isoform 3 lacks canonical residues 428–1023 and has a replacement at 404–427. An antibody recognizing a retained region could detect it, but the feature record does not show whether it is expressed in the sample. Check the antibody epitope and compare the band with an isoform-specific reference.
Which OGDH modifications matter when examining band changes?
PTM · The listed UniProt modifications are succinyllysine at 74 and 564, phosphoserine at 100, and acetyllysine at 401 and 970. These are UniProt coordinates; antibody or paper numbering may differ. Their presence alone does not establish a visible mobility shift. Confirm band identity before attributing a change to a modification.
Does this guide establish induction of OGDH?
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 OGDH?
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 A05301-1 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 can compartment choice affect OGDH quantitation?
Quantitation · OGDH localizes mainly to mitochondria, with a small nuclear fraction. Keep sample preparation consistent across comparisons and state whether measurements use whole-cell, mitochondrial, or nuclear material. A change in one fraction need not represent the same change in total OGDH.
Why is the observed OGDH band near 110 kDa?
Interpretation · The supplied apparent band is approximately 110 kDa, versus a predicted mass of 115.9 kDa. These features do not establish the cause of the difference. Use the observed band as a reference and check its identity before assigning another band to OGDH.
First compare them with the approximately 110 kDa observed band and the three annotated isoforms. Check epitope coverage, especially for isoform 3, which lacks residues 428–1023. The listed modifications do not, by themselves, identify an unexpected band or prove a visible shift.
First compare them with the approximately 110 kDa observed band and the three annotated isoforms. Check epitope coverage, especially for isoform 3, which lacks residues 428–1023. The listed modifications do not, by themselves, identify an unexpected band or prove a visible shift.
Boster reagents
OGDH 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-OGDH Antibody Picoband®
Cat # A05301-1
The catalog reports one anti-OGDH antibody for Western blot, A05301-1. Its blot caption reports a band near 110 kDa (expected 116 kDa) in the specified human cell and rat and mouse tissue lysates. Evidence is limited to the supplied product image and caption.
Which to pick: A05301-1 is the only listed option. It reports human, mouse, and rat reactivity and includes a Western blot image using 30 µg of lysate per lane and 0.5 µg/mL primary antibody. Match your sample and conditions to the caption.
Source: BosterBio OGDH gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q02218.
- Human Protein Atlas. OGDH tissue expression.
- PMC3116658 — target-verified WB comparison
- PMC12791878 — target-verified WB comparison
- PMC6752206 — target-verified WB comparison