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- Table of Contents
MT-CYB / Cytochrome b · Western blot design guide
Design a Western Blot for MT-CYB
Source-linked MT-CYB Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MT-CYB 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
MT-CYB Western Blot Experimental Design Guide
Expected bands, source-linked protocol options, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~42.7 kDa | |
| Observed band | ~43 kDa | |
| Gel | 10% (catalog A03737-2) | |
| Positive control | Parathyroid gland (IHC candidate; verify WB) +4 more | |
| Negative control | Bone marrow (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | — | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked MT-CYB Western Blot Protocol Options
The A03737-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human HepG2, human MDA-MB-231, human PC-3 (catalog A03737-2) |
| Gel % | 10% (catalog A03737-2) |
| Load | 30 ug; reducing conditions (catalog A03737-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A03737-2) |
| Membrane | nitrocellulose membrane (catalog A03737-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A03737-2) |
| Primary antibody | A03737-2 · 1:1000 (catalog A03737-2) |
| Primary incubation | overnight at 4°C (catalog A03737-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A03737-2) |
| Secondary incubation | 1.5 hour at RT (catalog A03737-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A03737-2) |
| Detection | ECL (catalog A03737-2) |
Section 2
What Is the Expected MT-CYB Western Blot Band Size?
MT-CYB is predicted at 42.7 kDa and observed near 43 kDa; the supplied evidence does not establish a cause for the small difference.
| Band near 43 kDa in whole-cell lysate | Matches the reported MT-CYB band and its 42.7 kDa predicted mass |
| Band near 43 kDa in a mitochondrial fraction | Consistent with MT-CYB in the mitochondrial inner membrane |
| Weak band near 43 kDa in whole-cell lysate | The inner-membrane protein may be poorly represented or recovered |
| Little or no band near 43 kDa in a cytosolic fraction | Consistent with mitochondrial inner-membrane localization |
💡Expected MT-CYB appearanceMT-CYB has a predicted mass of 42.7 kDa and an observed band at approximately 43 kDa in reducing whole-cell blots; confirm band identity with appropriate controls.
| Predicted molecular mass of 42.7 kDa | Agrees with the observed band near 43 kDa |
| 380-amino-acid sequence | Defines the reported full-length mass; no cleavage-related size change is listed |
| Mitochondrial inner-membrane localization | May affect recovery, with no size shift established |
| Multi-pass membrane topology | No distinct migration shift is established by the supplied evidence |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | MT-CYB is a mitochondrial inner-membrane protein | Check a mitochondrial fraction and confirm membrane-protein recovery |
| Weak or no signal | Poor recovery or transfer of this multi-pass membrane protein | Check solubilization, transfer, and a positive-control lysate |
| Band higher than expected | The listed features do not establish a higher-mass species | Compare with the reported 43 kDa band and verify identity using a positive control |
| Band lower than expected | No signal-peptide or propeptide cleavage is listed | Check sample integrity and verify the band with an independent antibody |
| Multiple bands | The supplied record lists no alternate isoforms or size-changing modifications | Identify the 43 kDa band using a positive control and assess other bands for nonspecific binding |
Experiment evidence
Western blot images showing MT-CYB band
A03737-2 · MT-CYB
A03737 · MT-CYB
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MT-CYB Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for MT-CYB in Western blot, you can use parathyroid gland tissue, where HPA reports high expression.
Positive control: Parathyroid gland (IHC candidate; verify WB)
Negative control: Bone marrow (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: As a multi-pass inner-mitochondrial-membrane protein, MT-CYB may require effective membrane solubilization for a clear signal.
HPA tissue expression evidence for MT-CYB
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 |
|---|---|---|---|---|
| Parathyroid gland | glandular cells | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | glandular 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 |
|---|---|---|---|---|
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Not detected | Protein (IHC) | HPA → |
| 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 → |
Section 3
Advanced MT-CYB Western Blot Tips
Deeper troubleshooting and optimisation questions for MT-CYB, answered from its protein features.
How should MT-CYB 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 an isoform explain another band?
Isoforms · The supplied UniProt record lists one isoform and no alternative sequence. It provides no isoform-based explanation for an additional band; check band identity before assigning one.
Do annotated modifications prove a band shift?
PTM · The linked UniProt record describes protein features. A modification annotation alone does not demonstrate a visible shift; retain any condition or experimental qualifier attached to it.
Does this guide establish induction of MT-CYB?
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 MT-CYB Western blot?
Transfer · MT-CYB is a multi-pass mitochondrial inner-membrane protein. The supplied features do not specify a transfer method. Check whether the approximately 43 kDa band transfers efficiently under your chosen conditions, including whether protein remains in the gel.
How should blocking be optimized for MT-CYB?
Blocking · The supplied features specify no preferred blocker. Compare blocking conditions using background near 43 kDa and the clarity of the target band as practical readouts.
How should MT-CYB bands be quantified?
Quantitation · Quantify the approximately 43 kDa band within the assay's linear range and use a consistent normalization method across samples. MT-CYB resides in the mitochondrial inner membrane, so changes in its signal can reflect changes in mitochondrial abundance as well as MT-CYB abundance per mitochondrion.
Should MT-CYB migrate at its predicted mass?
Interpretation · Its predicted mass is 42.7 kDa, and the supplied observed band is approximately 43 kDa. Their agreement supports the assignment, but apparent mass alone cannot establish band identity.
MT-CYB is part of the 11-subunit cytochrome bc1 complex and is a multi-pass membrane protein. Those features make association with other proteins relevant to investigate, but they do not establish the identity of a higher band. Confirm its identity experimentally rather than assigning it from size alone.
MT-CYB is part of the 11-subunit cytochrome bc1 complex and is a multi-pass membrane protein. Those features make association with other proteins relevant to investigate, but they do not establish the identity of a higher band. Confirm its identity experimentally rather than assigning it from size alone.
Boster reagents
MT-CYB 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-CYTB/MT-CYB Antibody
Cat # A03737-2
Real WB data
Anti-Cytochrome b MT-CYB Antibody
Cat # A03737
Both listed anti-MT-CYB antibodies have Western blot images. A03737-2 shows an approximately 43 kDa band in three human cell lysates; A03737 shows mouse tissue lanes. These images document those tested samples, not every listed species.
Which to pick: For human samples, A03737-2 has a human lysate image. For mouse tissues, A03737 has a mouse tissue image. Only A03737-2 lists rat reactivity, but neither supplied caption shows rat samples.
Source: BosterBio MT-CYB gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P00156.
- Human Protein Atlas. MT-CYB tissue expression.
- PMC7719573 — target-verified WB comparison
- PMC8418125 — target-verified WB comparison
- PMC12231110 — target-verified WB comparison