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- Table of Contents
ALB / Albumin · Western blot design guide
Design a Western Blot for ALB
Real validated ALB Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ALB 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
ALB 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 | ~69.4 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Kidney (IHC candidate; verify WB) | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Phosphorylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated ALB Western Blot Protocols
The A01245 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 % | 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 | A01245 · 1 μg/mL (catalog A01245) |
| Primary incubation | 1h (catalog A01245) |
| 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 ALB Western Blot Band Size?
Albumin’s predicted full-length mass is 69.4 kDa; secretion, cleavage, glycation, and isoforms may affect the blot, but their migration effects are not demonstrated here.
| Band near 69.4 kDa | Compatible with the predicted full-length precursor; identity requires controls. |
| Band below 69.4 kDa | Could reflect removal of the signal peptide and propeptide; the mature mass is not supplied. |
| Little or no band in whole-cell lysate | Albumin is secreted. |
| Several bands at different positions | Isoforms 1, 2, and 3 are annotated, but distinct migration is not established. |
💡Expected ALB appearanceThe predicted full-length mass is 69.4 kDa; secretion and precursor cleavage affect what may appear, but no empirical band size is supplied, so confirm identity with appropriate controls.
| Predicted full-length mass | 69.4 kDa is the sequence-based reference, not a measured band. |
| Signal peptide at residues 1–18 | Its removal makes the mature protein smaller than the precursor. |
| Propeptide at residues 19–24 | Its removal further reduces mass relative to the precursor. |
| Lysine 36 N-linked glucose glycation | May add a small amount of mass; a visible shift is not established. |
| Isoforms 1, 2, and 3 | May differ in size, but their masses and resolvable band positions are not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Albumin is secreted. | Check conditioned medium or a secretion-rich sample alongside a positive control. |
| Band higher than expected | Uncleaved precursor or a modified form is possible; the cause is unverified. | Compare with a positive control and verify antibody specificity. |
| Band lower than expected | Signal-peptide and propeptide cleavage produces a smaller mature protein. | Compare precursor and mature-compatible positions with a positive control. |
| Multiple bands | Isoforms 1, 2, and 3 are annotated, but their migration is unknown. | Check antibody specificity and compare bands across samples with a positive control. |
| Weak or no signal | Secreted albumin may be scarce in the sampled cell fraction. | Test medium or another suitable sample and confirm the antibody with a positive control. |
Experiment evidence
Western blot images showing ALB band
A01245 · ALB
A01245 · ALB
A01245 · ALB
M01245 · ALB
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ALB Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ALB in Western blot, you can use kidney tissue.
Positive control: Kidney (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside.
⚠️Feasibility: Because ALB is secreted, whole-cell lysates may show limited signal; consider an extracellular sample.
HPA tissue expression evidence for ALB
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 |
|---|---|---|---|---|
| Kidney | bowman's capsule | 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 | glandular 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 ALB Western Blot Tips
Deeper troubleshooting and optimisation questions for ALB, answered from its protein features.
What should be checked when unexpected ALB bands appear?
Band shift · Consider processing of the signal peptide (1–18) and propeptide (19–24), the deletions in isoforms 2 and 3, and antibody epitope coverage. ALB also has 17 listed disulfide bonds, so keep reducing conditions consistent when comparing lanes. The supplied features suggest checks, but do not identify any observed band.
How could ALB isoforms affect band interpretation?
Isoforms · UniProt lists three isoforms. Isoform 2 lacks positions 43–234, and isoform 3 lacks positions 164–376, using canonical precursor numbering. These deletions could yield smaller products. Check whether the antibody’s epitope lies in a deleted region before interpreting a missing or smaller band.
Which ALB phosphorylation sites matter when investigating multiple bands?
PTM · UniProt lists FAM20C phosphorylation at Ser29, Ser82, Ser89, and Thr107, plus phosphorylation at Ser297, Ser443, Thr444, Thr446, and Ser513. These are canonical precursor coordinates; antibody or paper numbering may differ. Their presence does not establish a visible shift. Compare samples under consistent preparation conditions before assigning a band to phosphorylation.
UniProt lists N-linked glycosylation at Asn342 in variant Redhill and Asn518 in variant Casebrook. These variant-specific sites should not be assumed present in every ALB sample. They are distinct from the listed lysine glycation sites. Confirm the variant before using either site to interpret a band.
UniProt lists N-linked glycosylation at Asn342 in variant Redhill and Asn518 in variant Casebrook. These variant-specific sites should not be assumed present in every ALB sample. They are distinct from the listed lysine glycation sites. Confirm the variant before using either site to interpret a band.
Does this guide establish induction of ALB?
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 ALB?
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 A01245 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 ALB bands be quantified across samples?
Quantitation · ALB is secreted, and UniProt lists three isoforms with different sequence deletions. Compare the same sample fraction and band definition across lanes. If measuring total ALB, check that the antibody recognizes the isoforms you intend to include; an epitope within positions 43–234 or 164–376 may be absent from an isoform.
Why might ALB migrate below its predicted 69.4 kDa?
Interpretation · The 69.4 kDa prediction is for the 609-residue precursor. ALB has a signal peptide at UniProt positions 1–18 and a propeptide at 19–24; processing removes these regions from mature secreted ALB. No empirical band position is supplied, so processing alone cannot establish the apparent mass of a particular band.
UniProt lists glycation at several lysines, including positions 36, 257, and 305; other sites, such as 75 and 161, are marked in vitro. Position 558 has alternate glycation and methylation annotations. Use canonical precursor numbering when checking sites. These annotations identify possible modifications, but do not demonstrate a visible band shift in your sample.
UniProt lists glycation at several lysines, including positions 36, 257, and 305; other sites, such as 75 and 161, are marked in vitro. Position 558 has alternate glycation and methylation annotations. Use canonical precursor numbering when checking sites. These annotations identify possible modifications, but do not demonstrate a visible band shift in your sample.
Boster reagents
ALB 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-Albumin Antibody
Cat # A01245
Real WB data
Anti-Albumin Rabbit Monoclonal Antibody
Cat # M01245
A01245 and M01245 both list human, mouse, and rat reactivity and have WB images. Their captions show human liver lysate for A01245 and rat and mouse blood and liver lysates for M01245. No publication evidence is supplied.
Which to pick: Choose A01245 for the documented human liver WB example (10 μg lysate; 1 μg/mL antibody). Choose M01245 for the rat or mouse blood and liver examples (30 μg; 1:500 overnight). Both have WB images; the captions do not demonstrate every listed reactivity.
Source: BosterBio ALB gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P02768.
- Human Protein Atlas. ALB tissue expression.
- PMC9299813 — target-verified WB comparison