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FKBP1A / Peptidyl-prolyl cis-trans isomerase FKBP1A · Western blot design guide
Design a Western Blot for FKBP1A
Source-linked FKBP1A Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-FKBP1A 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
FKBP1A 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 | ~12 kDa | |
| Gel | 15% (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 | Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked FKBP1A Western Blot Protocol Options
The A04492 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | A431 cell lysate (catalog A04492) |
| Gel % | 15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.2 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A04492 · 1 μg/ml (catalog A04492) |
| 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 FKBP1A Western Blot Band Size?
FKBP1A is predicted near 12 kDa; alternate Lys53 modifications are listed, but their effects on migration are unproven.
| Single band near 12 kDa | consistent with the predicted full-length FKBP1A size; confirm identity with an antibody control |
| Band near 12 kDa in cytosolic fraction | consistent with cytosolic FKBP1A |
| Band near 12 kDa in sarcoplasmic reticulum membrane fraction | consistent with its reported peripheral membrane location |
| Close doublet near 12 kDa | cannot be assigned to alternate Lys53 modifications without validation |
💡Expected FKBP1A appearanceUniProt predicts FKBP1A near 12 kDa, but no empirical band size is supplied; confirm a candidate band with antibody specificity or FKBP1A depletion controls.
| UniProt predicted mass | places the full-length band near 12 kDa |
| 108-residue sequence | underlies the predicted full-length mass; no cleavage is listed |
| Alternate Lys53 N6-acetylation | is listed, but a visible mobility change is not established |
| Alternate Lys53 N6-succinylation | is listed, but a visible mobility change is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | cytosolic FKBP1A may have been lost during fractionation | check the cytosolic fraction and a positive lysate control |
| Band higher than expected | identity is uncertain; no glycosylation or disulfide feature explains a large shift | verify specificity with FKBP1A depletion or a second antibody |
| Band lower than expected | possible degradation or nonspecific binding; no cleavage feature is listed | use fresh lysate with protease inhibitors and verify band identity |
| Multiple bands | one isoform is listed; alternate Lys53 modifications do not establish distinct bands | compare bands after FKBP1A depletion |
| Weak or no signal | poor recovery of cytosolic FKBP1A or low assay sensitivity | check cytosolic extraction and a positive lysate control |
Experiment evidence
Western blot images showing FKBP1A band
A04492 · FKBP1A
M04492 · FKBP1A
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for FKBP1A Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for FKBP1A in Western blot, you can use adrenal gland tissue, which HPA rates as medium expression.
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 alongside the samples.
⚠️Feasibility: Cytosolic FKBP1A should be accessible in tissue lysate, with HPA not-detected adipose tissue as a potential negative control.
HPA tissue expression evidence for FKBP1A
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 | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Caudate | neuronal cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal 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 → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cervix | glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced FKBP1A Western Blot Tips
Deeper troubleshooting and optimisation questions for FKBP1A, answered from its protein features.
How should FKBP1A 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.
Are multiple FKBP1A isoforms expected?
Isoforms · Only one isoform is listed, with no alternative sequence supplied. The supplied features therefore do not support assigning separate bands to different FKBP1A isoforms.
Could modifications at residue 53 affect the FKBP1A band?
PTM · UniProt lists alternate N6-acetyllysine and N6-succinyllysine at position 53. These are UniProt coordinates; antibody or paper numbering may differ. Their presence alone does not establish a visible band shift.
Does this guide establish induction of FKBP1A?
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 FKBP1A Western blot?
Transfer · FKBP1A is small, with a predicted mass of 12 kDa. Choose a transfer setup suitable for retaining small proteins and check transfer using a low-mass marker. The supplied features do not specify a particular transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A04492 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 FKBP1A bands be quantified?
Quantitation · Quantify a consistently identified band near the 12 kDa reference and keep the measurement within the assay’s linear range. Because no empirical band position is supplied, confirm band identity before including additional bands in the measurement.
Where should FKBP1A run relative to its predicted mass?
Interpretation · The predicted mass is 12 kDa. No observed band position is supplied, so use 12 kDa as a reference rather than an expected exact position. The listed modifications do not establish a visible shift.
The features list one isoform, no signal or propeptide, no glycosylation sites, and alternate modifications at residue 53. They do not establish the cause of an additional band. Compare its position with the 12 kDa reference and verify its identity before assigning it to FKBP1A.
The features list one isoform, no signal or propeptide, no glycosylation sites, and alternate modifications at residue 53. They do not establish the cause of an additional band. Compare its position with the 12 kDa reference and verify its identity before assigning it to FKBP1A.
Boster reagents
FKBP1A 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-FKBP12 FKBP1A Antibody
Cat # A04492
Real WB data
Anti-FKBP12 Rabbit Monoclonal Antibody
Cat # M04492
A04492 and M04492 are anti-FKBP1A (FKBP12) antibodies listing human, mouse, and rat reactivity. Both have WB images. Their captions document specific lysate examples; M04492 reports a band at approximately 12 kDa. Broader specimen performance is not established by these examples.
Which to pick: A04492 has a WB example in A431 lysate at 1 μg/ml. M04492 has a WB example using specified human, rat, and mouse lysates at 1:1000. Choose based on the reported samples and conditions closest to your experiment.
Source: BosterBio FKBP1A gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P62942.
- Human Protein Atlas. FKBP1A tissue expression.
- PMC11889056 — target-verified WB comparison
- PMC8726715 — target-verified WB comparison