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- Table of Contents
PRDM16 / Histone-lysine N-methyltransferase PRDM16 · Western blot design guide
Design a Western Blot for PRDM16
Real validated PRDM16 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PRDM16 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
PRDM16 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 | ~140.3 kDa | |
| Gel | 8–10% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 4 isoform(s) |
Section 1
Real Curated PRDM16 Western Blot Protocols
The A00782 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Target-positive lysate and matched negative control (standard starting point) |
| 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 | A00782; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | 1 h (catalog A00782) |
| 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 PRDM16 Western Blot Band Size?
PRDM16 is predicted at 140.3 kDa; four isoforms could affect migration, but no empirical band position or distinct isoform bands are established.
| Band near 140.3 kDa | Consistent with the predicted PRDM16 mass; identity requires confirmation |
| Several bands at different positions | Could reflect PRDM16 isoforms 1, 2, 3, and 4 if they migrate differently |
| One band despite four annotated isoforms | The isoforms may not resolve or may not all be detected |
| Weak or absent band in whole-cell lysate | PRDM16 is found in the nucleus, on chromosomes, and in the cytoplasm |
💡Expected PRDM16 appearancePRDM16 has a predicted mass of 140.3 kDa, but no empirical band position or isoform migration is supplied; confirm any candidate band with ordinary identity controls.
| Predicted PRDM16 mass | Provides a 140.3 kDa sequence-based reference, not a measured band position |
| Isoform 1 | Its individual mass and migration are not supplied |
| Isoform 2 | May differ in size from other isoforms; its mass is not supplied |
| Isoform 3 | May differ in size from other isoforms; its mass is not supplied |
| Isoform 4 | May differ in size from other isoforms; its mass is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear or chromosome-associated PRDM16 may be poorly recovered | Check nuclear extraction and use a positive control |
| Band higher than expected | The reason for migration above 140.3 kDa is not established | Check marker calibration and confirm identity by knockdown or a second antibody |
| Band lower than expected | An alternative isoform is possible, but its mass is unknown | Check antibody epitope coverage and confirm identity by knockdown |
| Multiple bands | Four annotated isoforms could contribute if expressed and resolved | Compare isoform expression and use knockdown to identify PRDM16 bands |
| Weak or no signal | PRDM16 distribution between nucleus and cytoplasm may affect recovery | Compare nuclear and cytoplasmic fractions with loading controls |
Experiment evidence
Western blot images showing PRDM16 band
A00782 · PRDM16
A00782 · PRDM16
A00782 · PRDM16
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PRDM16 Western blot
🧪For positive controls for PRDM16 in Western blot, you can use no HPA-validated tissue or cell line from the supplied evidence.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA expression data are unavailable, so tissue-based controls cannot be validated from this evidence.
HPA tissue expression evidence for PRDM16
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced PRDM16 Western Blot Tips
Deeper troubleshooting and optimisation questions for PRDM16, answered from its protein features.
How should PRDM16 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 PRDM16 isoforms affect band interpretation?
Isoforms · Four isoforms are listed. Relative to the canonical sequence, isoform 4 lacks residues 1–184, isoform 2 lacks 1233–1251, and isoform 3 has changes at 191 and 868. Check whether the antibody recognizes a region retained in the isoform being measured.
Could acetylation change the PRDM16 band?
PTM · UniProt lists alternate N6-acetyllysine at position 915, using canonical sequence numbering. This identifies a modification site, but does not establish a visible band shift. Check the numbering convention before comparing it with an antibody or paper.
Does this guide establish induction of PRDM16?
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 PRDM16 Western blot?
Transfer · PRDM16 has a predicted mass of 140.3 kDa. Check transfer of proteins around that size with a suitable marker and, if signal is weak, inspect the gel for retained protein. The supplied features do not specify a transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A00782 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 PRDM16 bands be quantified?
Quantitation · Define which isoform or bands the antibody detects before quantifying. Compare equivalent sample fractions across lanes: PRDM16 is reported in the nucleus and cytoplasm, and also at chromosomes. The features do not establish that every band represents the same protein form.
Should PRDM16 run at its predicted 140.3 kDa?
Interpretation · Use 140.3 kDa as the predicted mass of the canonical protein. No observed band position is supplied, and the listed features do not establish an apparent mass or explain a shift.
Consider the listed sequence variants: isoform 4 lacks canonical residues 1–184, isoform 2 lacks 1233–1251, and isoform 3 lacks residue 868. An antibody whose binding region is absent may miss an isoform. Band size alone cannot identify it.
PRDM16 is reported in the nucleus and cytoplasm, with chromosome localization also listed. Compare matched fractions and check fraction purity when interpreting a weak or unexpected signal. Localization alone does not establish the amount expected in each fraction.
Consider the listed sequence variants: isoform 4 lacks canonical residues 1–184, isoform 2 lacks 1233–1251, and isoform 3 lacks residue 868. An antibody whose binding region is absent may miss an isoform. Band size alone cannot identify it.
PRDM16 is reported in the nucleus and cytoplasm, with chromosome localization also listed. Compare matched fractions and check fraction purity when interpreting a weak or unexpected signal. Localization alone does not establish the amount expected in each fraction.
Boster reagents
PRDM16 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-PRDM16 Antibody
Cat # A00782
The catalog reports A00782, an anti-PRDM16 antibody with reported Human, Mouse, and Rat reactivity. Its Western blot image uses 15 μg of MCF7 cell lysate; the supplied evidence does not show Western blot results for other specimens.
Which to pick: A00782 is the only listed option. Choose it if its reported reactivity fits your sample; its supplied Western blot image documents MCF7 cells under the stated conditions, so performance in other samples remains unshown here.
Source: BosterBio PRDM16 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9HAZ2.
- Human Protein Atlas. PRDM16 tissue expression.
- PMC10870028 — target-verified WB comparison
- PMC11965059 — target-verified WB comparison
- PMC4575949 — target-verified WB comparison