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FKBP5 / Peptidyl-prolyl cis-trans isomerase FKBP5 · Western blot design guide
Design a Western Blot for FKBP5
Real validated FKBP5 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-FKBP5 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
FKBP5 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 | ~51.2 kDa | |
| Observed band | ~51 kDa | |
| Gel | 5–20% (catalog M04182) | |
| Positive control | Adipose tissue (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 | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated FKBP5 Western Blot Protocols
The M04182 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human CACO-2, human Hela, human Jurkat (catalog M04182) |
| Gel % | 5–20% (catalog M04182) |
| Load | 30 ug; reducing conditions (catalog M04182) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M04182) |
| Membrane | nitrocellulose membrane (catalog M04182) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M04182) |
| Primary antibody | M04182 · 1:500 (catalog M04182) |
| Primary incubation | overnight at 4°C (catalog M04182) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:500 (catalog M04182) |
| Secondary incubation | 1.5 hour at RT (catalog M04182) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M04182) |
| Detection | ECL (catalog M04182) |
Section 2
What Is the Expected FKBP5 Western Blot Band Size?
FKBP5 is predicted at 51.2 kDa and observed at ~51 kDa; the cause of the small difference is not established.
| Band at ~51 kDa | Matches the reported FKBP5 band in reducing whole-cell lysates |
| Single band near 51 kDa | Consistent with the predicted 51.2 kDa size; two isoforms need not resolve separately |
| Additional band at a different position | Could reflect isoform 1 or 2, but their individual migration is unknown |
| Band in a nuclear or cytoplasmic fraction | Consistent with FKBP5 localization in both compartments |
💡Expected FKBP5 appearanceFKBP5 is predicted at 51.2 kDa and observed at ~51 kDa in reducing whole-cell blots; confirm band identity with a positive lysate and an independent antibody.
| UniProt predicted mass | 51.2 kDa predicted; the reported whole-cell band is ~51 kDa |
| Splice isoform 1 | Its individual apparent size is not supplied |
| Splice isoform 2 | Its individual apparent size is not supplied |
| Alternative splicing of isoforms 1 and 2 | Could change apparent size, but distinct migration is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | FKBP5 signal may be below detection in the sampled lysate | Check loading and detection with a known positive whole-cell lysate |
| Band higher than expected | An isoform or unrelated antibody target could migrate higher; identity is unproven | Compare with a positive lysate and an independent FKBP5 antibody |
| Band lower than expected | An isoform or unrelated antibody target could migrate lower; identity is unproven | Compare with a positive lysate and an independent FKBP5 antibody |
| Multiple bands | Two splice isoforms exist, but separate band positions are unreported | Check each band with an independent antibody or isoform-specific control |
| Weak or no signal | FKBP5 distribution between cytoplasm and nucleus may affect fraction signal | Check both fractions and verify loading and detection |
Experiment evidence
Western blot images showing FKBP5 band
M04182 · FKBP5
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for FKBP5 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for FKBP5 in Western blot, you can use adipose tissue, which HPA rates as high expression.
Positive control: Adipose tissue (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: No supplied tissue is rated not detected, so use siRNA knockdown or a KO line for a clear negative control.
HPA tissue expression evidence for FKBP5
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 |
|---|---|---|---|---|
| Adipose tissue | adipocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Breast | adipocytes | Medium | Protein (IHC) | HPA → |
| Caudate | glial cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | endothelial cells | Medium | Protein (IHC) | HPA → |
| Duodenum | glandular cells | Medium | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced FKBP5 Western Blot Tips
Deeper troubleshooting and optimisation questions for FKBP5, answered from its protein features.
How should FKBP5 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 FKBP5 isoforms produce different bands?
Isoforms · Two isoforms are listed. In isoform 2, residues 223–268 are replaced by a different sequence and residues 269–457 are missing. An antibody’s recognition of either isoform depends on its epitope; check that before assigning a smaller band.
Isoform 2 lacks canonical residues 269–457, including phosphoserine 445. Check the antibody’s isoform specificity before interpreting a phosphoserine 445 signal. These coordinates use the supplied canonical UniProt sequence numbering.
Isoform 2 lacks canonical residues 269–457, including phosphoserine 445. Check the antibody’s isoform specificity before interpreting a phosphoserine 445 signal. These coordinates use the supplied canonical UniProt sequence numbering.
Which FKBP5 modifications matter when interpreting a blot?
PTM · The supplied UniProt coordinates list N-acetylmethionine at 1, phosphoserine at 13 and 445, and N6-acetyllysine at 28 and 155. These features alone cannot establish which modifications are present in a sample or whether they alter migration.
Does this guide establish induction of FKBP5?
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 FKBP5?
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 M04182 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 FKBP5 be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
Why is the FKBP5 band near 51 kDa?
Interpretation · The canonical sequence has a predicted mass of 51.2 kDa, consistent with the reported apparent band near 51 kDa. The listed modifications do not, by themselves, establish a visible shift.
FKBP5 is listed in both cytoplasm and nucleus. When comparing fractions, quantify each against an appropriate loading reference for that fraction, and consider whether the antibody recognizes both isoforms.
The supplied complex description says FKBP5 dissociates from a complex containing HSP90, HSPA1A/HSPA1B and steroid receptors upon ligand binding, with FKBP4 taking its place. That describes a change in complex membership, not an increase in total FKBP5 abundance.
Check antibody epitope coverage against isoform 2’s replacement of residues 223–268 and loss of 269–457. The listed modifications do not independently explain an unexpected band’s position; feature annotations alone cannot identify that band.
FKBP5 is listed in both cytoplasm and nucleus. When comparing fractions, quantify each against an appropriate loading reference for that fraction, and consider whether the antibody recognizes both isoforms.
The supplied complex description says FKBP5 dissociates from a complex containing HSP90, HSPA1A/HSPA1B and steroid receptors upon ligand binding, with FKBP4 taking its place. That describes a change in complex membership, not an increase in total FKBP5 abundance.
Check antibody epitope coverage against isoform 2’s replacement of residues 223–268 and loss of 269–457. The listed modifications do not independently explain an unexpected band’s position; feature annotations alone cannot identify that band.
Boster reagents
FKBP5 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-FKBP51 Monoclonal Antibody
Cat # M04182
The listed anti-FKBP5 antibody, M04182, has a Western blot image showing an approximately 51 kDa band in human cell and rat tissue or cell lysates. Evidence is limited to the samples and conditions reported in that image.
Which to pick: M04182 is the only listed option. Its reported human and rat reactivity is supported by a WB image using human CACO-2, HeLa, Jurkat and HepG2 cells, rat thymus and PC-12 cells, with 30 µg lysate and a 1:500 primary dilution.
Source: BosterBio FKBP5 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q13451.
- Human Protein Atlas. FKBP5 tissue expression.
- PMC8726715 — target-verified WB comparison
- PMC8108082 — target-verified WB comparison
- PMC10349081 — target-verified WB comparison