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- Table of Contents
PC / Pyruvate carboxylase, mitochondrial · Western blot design guide
Design a Western Blot for PC
Real validated PC Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PC 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
PC 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 | ~129.6 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Adrenal gland (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 | Phosphorylated + Acetylated | |
| Caveat | Alternative splicing | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated PC Western Blot Protocols
The M03853 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | HepG2 cell lysate (catalog M03853) |
| Gel % | 8–10% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Wet/tank transfer; optimize duration (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M03853; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected PC Western Blot Band Size?
PC has a predicted 129.6 kDa monomer; mitochondrial transit peptide removal may lower the mature band, and isoforms 1 and 2 may yield additional bands.
| Band near 129.6 kDa | consistent with the PC monomer's predicted mass |
| Band below 129.6 kDa | may reflect removal of the mitochondrial transit peptide |
| Bands at different sizes | may reflect PC isoforms 1 and 2 |
| Faint band in whole-cell lysate | PC is located in the mitochondrial matrix |
💡Expected PC appearanceExpect a PC monomer band near the predicted 129.6 kDa, potentially somewhat lower after mitochondrial transit peptide removal, with additional bands possible from isoforms 1 and 2.
| UniProt predicted mass | places the full-length PC monomer near 129.6 kDa |
| Mitochondrial transit peptide | its removal may make mature PC smaller than the full-length protein |
| Isoform 1 | may differ in size from isoform 2 |
| Isoform 2 | may differ in size from isoform 1 |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | mitochondrial matrix PC may be scarce in the sampled lysate | check a mitochondria-enriched fraction |
| Band lower than expected | removal of the mitochondrial transit peptide | compare the band with a mitochondria-enriched fraction |
| Multiple bands | PC isoforms 1 and 2 may differ in size | check which isoforms the antibody recognizes |
| Weak or no signal | low PC abundance in the sampled fraction | increase lysate input or enrich mitochondria |
| Fragments below expected size | PC degradation during sample preparation | prepare fresh lysate with protease inhibitors |
Experiment evidence
Western blot images showing PC band
M03853 · PC
A03853-2 · PC
A03853-3 · PC
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PC Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PC in Western blot, you can use adrenal gland lysate.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: PC is mitochondrial, and HPA lists adipose tissue as not detected, so tissue lysates offer a feasible positive and negative pair.
HPA tissue expression evidence for PC
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 → |
| Cerebellum | cells in granular layer | High | Protein (IHC) | HPA → |
| Cervix | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | 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 → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | Not detected | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PC Western Blot Tips
Deeper troubleshooting and optimisation questions for PC, answered from its protein features.
Where should the main PC band appear?
Band shift · PC has a predicted mass of 129.6 kDa. Use that as a starting point, then confirm any candidate band with appropriate controls; no observed band position is supplied.
Consider the two listed isoforms and PC's modified residues, but neither establishes the identity of an unexpected band. PC is a mitochondrial matrix protein with a transit-peptide keyword; the supplied features give no cleavage site or mature mass. Validate band identity before assigning a cause.
Consider the two listed isoforms and PC's modified residues, but neither establishes the identity of an unexpected band. PC is a mitochondrial matrix protein with a transit-peptide keyword; the supplied features give no cleavage site or mature mass. Validate band identity before assigning a cause.
Could PC isoforms produce more than one band?
Isoforms · UniProt lists two alternatively spliced isoforms, 1 and 2. They may contribute to band differences, but their individual masses are not supplied. Do not assign a band to either isoform from position alone.
Could PC modifications affect band migration?
PTM · PC has 24 modified-residue annotations and is associated with acetylation, phosphorylation, and biotin. These features can complicate band interpretation; the supplied data do not establish a specific shift or treatment-dependent change.
Does this guide establish induction of PC?
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 PC Western blot?
Transfer · PC is predicted to be 129.6 kDa, so verify that your transfer conditions recover proteins near 130 kDa. Check transfer efficiency before interpreting a weak or absent PC band; the supplied features do not specify one transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M03853 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 PC be quantified?
Quantitation · Quantify a validated PC band near its predicted 129.6 kDa mass. PC is located in the mitochondrial matrix, so compare samples prepared with consistent mitochondrial recovery and loading. Keep any additional bands separate until their identity is established.
What confirms that a band represents PC?
Interpretation · Use target-positive and orthogonal negative controls such as knockout or knockdown where feasible. HPA tissue IHC candidates require WB validation. Do not assign an unexpected band solely by apparent mass.
Boster reagents
PC 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-PCB Rabbit Monoclonal Antibody
Cat # M03853
Real WB data
Anti-Pyruvate Carboxylase/PC Antibody
Cat # A03853-2
Real WB data
Anti-Pyruvate Carboxylase/PC Antibody Picoband®
Cat # A03853-3
Three the supplier antibodies are listed for PC Western blotting, and each has a WB image. M03853 shows PCB expression in human HepG2 cell lysate. A03853-2 shows human HeLa, HepG2, MCF-7, and SiHa cell lysates; rat liver and RH-35 lysates; and mouse liver and Hepa1-6 lysates. A03853-3 shows human HepG2, HeLa, and SiHa cell lysates; rat liver and RH35 lysates; and mouse liver lysate. No publication evidence is supplied.
Which to pick: All three show HepG2 samples. For the other listed samples, compare the A03853-2 and A03853-3 captions above; MCF-7 and Hepa1-6 appear only with A03853-2. These examples do not establish broader species reactivity or comparative performance.
Source: BosterBio PC gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P11498.
- Human Protein Atlas. PC tissue expression.
- PMC11967762 — target-verified WB comparison