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PDZK1 / Na(+)/H(+) exchange regulatory cofactor NHE-RF3 · Western blot design guide
Design a Western Blot for PDZK1
Source-linked PDZK1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PDZK1 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
PDZK1 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 | ~57.1 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Duodenum (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 | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked PDZK1 Western Blot Protocol Options
The M03176-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | T47-D cell lysate (catalog M03176-1) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M03176-1; 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 PDZK1 Western Blot Band Size?
PDZK1 is predicted at 57.1 kDa; isoforms and phosphorylation could affect migration, but no empirical band or feature-dependent shift is established.
| Band near 57.1 kDa | Compatible with the predicted PDZK1 mass; confirm identity with controls |
| Additional band at a different size | Could reflect isoform 1 or 2; their migration is not established |
| Slightly shifted band or doublet | Could reflect phosphorylation; a visible shift is not established |
| Weak band in a soluble fraction | PDZK1 associates with peripheral membranes |
💡Expected PDZK1 appearancePDZK1 has a predicted mass of 57.1 kDa, but no empirical band size is supplied; confirm any candidate band with antibody specificity or PDZK1 depletion controls.
| Predicted PDZK1 mass | Places the reference band near 57.1 kDa |
| Isoform 1 | May migrate differently from isoform 2; its mass is not supplied |
| Isoform 2 | May migrate differently from isoform 1; its mass is not supplied |
| Documented phosphoserine and phosphothreonine sites | Could affect migration, but no visible size shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Peripheral membrane association or insufficient PDZK1 in the sample | Check a membrane-enriched fraction and verify sample expression |
| Band higher than expected | Phosphorylation is documented, but the band identity and cause of the shift are unknown | Compare phosphatase-treated material and confirm PDZK1 with depletion controls |
| Band lower than expected | An isoform or protein breakdown may account for a smaller band | Compare isoform expression and confirm the band with PDZK1 depletion controls |
| Multiple bands | Two isoforms or phosphorylated forms are possible; distinct migration is unverified | Use PDZK1 depletion and phosphatase treatment to identify the bands |
| Weak or no signal | PDZK1 may be poorly recovered from membrane-associated material | Check extraction and sample loading with a suitable control |
Experiment evidence
Western blot images showing PDZK1 band
M03176-1 · PDZK1
A03176-1 · PDZK1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PDZK1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PDZK1 in Western blot, you can use duodenum lysate.
Positive control: Duodenum (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: PDZK1 is membrane-associated, so membrane-enriched lysate may improve detection.
HPA tissue expression evidence for PDZK1
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 |
|---|---|---|---|---|
| Duodenum | endocrine cells | High | Protein (IHC) | HPA → |
| Kidney | bowman's capsule | High | Protein (IHC) | HPA → |
| Small intestine | endocrine cells | High | Protein (IHC) | HPA → |
| Colon | mucosal lymphoid cells | Medium | Protein (IHC) | HPA → |
| Gallbladder | glandular cells | Medium | 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 → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | endocrine cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PDZK1 Western Blot Tips
Deeper troubleshooting and optimisation questions for PDZK1, answered from its protein features.
How should PDZK1 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 PDZK1 isoforms affect band interpretation?
Isoforms · UniProt lists isoforms 1 and 2. Isoform 2 lacks canonical residues 154–264, so it may produce a smaller band if expressed and recognized by the antibody. Check whether the antibody epitope lies within that missing segment. These coordinates use the supplied canonical sequence numbering.
Which PDZK1 phosphorylation sites should a blot comparison consider?
PTM · The supplied UniProt coordinates list phosphoserine at 108, 148, 192, 250, 334, 348, 492, 508, 510, 511, 512 and 514, plus phosphothreonine at 451. These are canonical UniProt positions; antibody or paper numbering may differ. Site annotations alone do not show which residues are phosphorylated in a particular sample.
Canonical PDZK1 has phosphoserine annotations at 508, 510, 511, 512 and 514. Multiple bands could motivate a phosphorylation comparison, but the annotations do not demonstrate a visible mobility shift. Compare matched samples with and without phosphatase treatment to assess whether phosphorylation contributes.
Canonical PDZK1 has phosphoserine annotations at 508, 510, 511, 512 and 514. Multiple bands could motivate a phosphorylation comparison, but the annotations do not demonstrate a visible mobility shift. Compare matched samples with and without phosphatase treatment to assess whether phosphorylation contributes.
Does this guide establish induction of PDZK1?
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 PDZK1 Western blot?
Transfer · PDZK1 is a peripheral membrane protein with a predicted canonical mass of 57.1 kDa. The supplied features do not specify a transfer method. Validate transfer around that mass by checking signal retained in the gel and signal captured on the membrane.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M03176-1 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 should PDZK1 localization guide quantitative comparisons?
Quantitation · PDZK1 associates with peripheral membranes and is reported at the cell membrane, including the apical compartment of proximal tubular cells and sinusoidal liver membranes. Compare equivalent sample fractions across conditions; changes in fraction recovery could alter band intensity without establishing a change in total PDZK1.
Why might PDZK1 migrate away from its predicted 57.1 kDa?
Interpretation · Use 57.1 kDa as the predicted mass of canonical PDZK1, not an expected observed band position. The listed phosphorylation sites and alternative splicing warrant investigation, but their presence alone does not establish a visible shift or explain any mass difference. No observed band position was supplied.
Check the antibody epitope against isoform 2’s missing canonical residues 154–264, then compare band behavior after phosphatase treatment. PDZK1’s listed interactions do not by themselves identify an extra band. Without an observed band position, the supplied features cannot assign a specific unexpected band.
Check the antibody epitope against isoform 2’s missing canonical residues 154–264, then compare band behavior after phosphatase treatment. PDZK1’s listed interactions do not by themselves identify an extra band. Without an observed band position, the supplied features cannot assign a specific unexpected band.
Boster reagents
PDZK1 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-PDZK1 Rabbit Monoclonal Antibody
Cat # M03176-1
Real WB data
Anti-PDZK1 Antibody Picoband®
Cat # A03176-1
Both listed anti-PDZK1 antibodies have Western blot images. M03176-1 was shown with T47-D lysate; A03176-1 was shown with human cell and rat and mouse tissue lysates. A03176-1 detected a band near 70 kDa, above the stated 57 kDa expectation.
Which to pick: For human T47-D lysate, consider M03176-1. For the listed human cell lines or rat and mouse tissues, A03176-1 has the more directly relevant image. Check the observed 70 kDa band against your expected result.
Source: BosterBio PDZK1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q5T2W1.
- Human Protein Atlas. PDZK1 tissue expression.
- PMC10964575 — target-verified WB comparison
- PMC12644722 — target-verified WB comparison
- PMC4504350 — target-verified WB comparison