ALB / Albumin · IHC design guide

Design Immunohistochemistry for ALB

Plan ALB IHC-P around plasma and extracellular matrix positivity (HPA tissue IHC). The A01245 human liver example uses formaldehyde fixation, 2 μg/mL primary antibody, overnight incubation at 4 °C, HRP detection and hematoxylin counterstain (datasheet A01245 IHC-P).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use

Immunohistochemistry protocol sheet for ALB (IHC for ALB): expected localisation Plasma and extracellular matrix (HPA tissue IHC), antibody A01245, validated IHC image, and IHC protocol steps — Printable **ALB** IHC protocol sheet — expected localisation Plasma and extracellular matrix (HPA tissue IHC), antibody A01245, controls and protocol steps. Open the full ALB IHC guide →

Navigate to: Protocols Staining pattern Controls Antibodies

ALB Immunohistochemistry Experimental Design Guide

Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.

Must know before staining

	Expected localisation	Plasma and extracellular matrix (HPA tissue IHC)
	Staining pattern	Plasma and extracellular matrix positivity (HPA tissue IHC)
	Antigen retrieval	Citrate pH 6 HIER, heat-mediated (datasheet A01245)
	Positive control ⓘ	Kidney
	Negative control ⓘ	Adipose tissue+4 more · see all

Important caveats

Reasons your staining may differ from the expected pattern.

	Fixation	Formaldehyde-fixed paraffin sections (datasheet A01245 IHC-P) (selected-SKU IHC image A01245)
	Caveat	Secreted ALB can make tissue RNA and protein differ (HPA tissue IHC)
	Regulation	Liver-enriched RNA (HPA tissue RNA)
	Isoform / epitope	3 isoforms; signal peptide and propeptide are cleaved, so check epitope coverage (UniProt)

Section 1

Recommended ALB IHC & IF Protocols

The catalog antibody has an IHC-P protocol (datasheet A01245). Two published ALB IHC methods provide conditions for quail UVJ mucosa and liver sections (PMC7063080; PMC13149365).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Formaldehyde-fixed, paraffin-embedded human liver tissue (datasheet A01245)
Fixation	Image formalin-fixed; duration unreported (datasheet A01245); verify before use.
Sectioning	4–5 µm sections on charged slides (standard)
Deparaffinisation	Xylene, graded ethanol series to water (standard)
Antigen retrieval	Heat retrieval: Citrate pH 6 (datasheet A01245); 20 min, 95–100 °C (standard)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% serum (datasheet A01245)
Primary antibody	Chicken anti-ALB, 10-20 μg/mL (datasheet A01245)
Primary incubation	Overnight at 4 °C (standard)
Detection	HRP-polymer secondary, DAB chromogen 5–10 min (standard)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	ALB-positive staining in bowman's capsule of kidney (HPA tissue IHC: Medium). HPA tissue profile: Positivity in plasma and extracellular matrix. No signal in the no-primary control.

💡Decision noteStart with citrate pH 6 HIER for the catalog antibody (datasheet A01245). Neither published excerpt specifies antigen retrieval (PMC7063080; PMC13149365).

Section 2

What Is the Expected ALB Staining Pattern?

ALB is secreted and has no transmembrane segment (UniProt P02768: subcellular location and topology). In paraffin-section IHC, expect staining in plasma and extracellular matrix (HPA tissue IHC: Enhanced reliability; reported profile). Bowman's capsule is a documented medium-positive kidney site (HPA tissue IHC: Kidney). Liver-enriched RNA does not establish a corresponding cellular IHC pattern because secreted protein and RNA locations can differ (HPA tissue IHC: reliability description).

What am I looking at on my slide?
Plasma and extracellular matrix stain, with medium signal at Bowman's capsule.	This fits the reported IHC profile and kidney annotation (HPA tissue IHC: plasma and extracellular matrix; Bowman's capsule, Medium). Judge the kidney signal in its annotated location; HPA does not provide a universal intensity threshold for every tissue.
Strong, sharply nuclear or membrane-restricted staining dominates.	Those compartments conflict with secreted ALB and its lack of a transmembrane segment (UniProt P02768: location and topology). Review the slide pattern and controls before assigning such staining to ALB; intracellular signal requires separate context.
Adipocytes or respiratory epithelial cells stain strongly.	These cell types were not detected in the supplied HPA tissue IHC observations (HPA tissue IHC: adipose tissue and bronchus). Consider nonspecific antibody binding or chromogenic detection activity; a discrepant result alone cannot identify its cause (standard IHC practice).
Color spreads across the section without resolving plasma or matrix structures.	Diffuse signal is hard to distinguish from nonspecific background when the expected extracellular compartment is broad (HPA tissue IHC: profile). Compare morphology and negative controls, then assess blocking, washing and detection conditions (standard IHC practice).
Bowman's capsule is unstained in a technically adequate kidney section.	This differs from the supplied medium-positive observation (HPA tissue IHC: Kidney, Bowman's capsule). First check antibody and detection performance with controls; a negative section alone does not prove absence of ALB (standard IHC practice).

💡Expected ALB appearanceCall positive IHC when plasma or extracellular matrix is convincingly stained and kidney Bowman's capsule shows its reported medium signal; isolated strong nuclear staining or strong signal in HPA-negative cell types warrants investigation (HPA tissue IHC: profile and kidney level; UniProt P02768: secreted location).

How each factor affects the staining
Secretion and processing	ALB has a signal peptide at residues 1–18, a propeptide at 19–24 and a mature chain at 25–609 (UniProt P02768: processing). An antibody's epitope determines which forms it can recognize; no epitope is supplied here.
Tissue RNA versus protein	RNA is liver-enriched, while the supplied IHC profile reports plasma and extracellular matrix positivity (HPA tissue IHC: RNA specificity and profile). HPA cautions that their tissue locations may differ for secreted variants (HPA tissue IHC: reliability description).
Antibody validation	Two listed rabbit antibodies have Enhanced IHC status; a listed mouse antibody has Supported IHC status (HPA antibodies: HPA031024, HPA031025, CAB006262). These ratings describe those antibodies and do not validate an unspecified catalog antibody.
Observed negative cell types	HPA reports no detection in adipocytes, respiratory epithelial cells and several other listed cell types (HPA tissue IHC: negative observations). Use a matched negative tissue to assess unexpected cellular staining, while checking its morphology and section quality (standard IHC practice).
IF/ICC Q&A: where can intracellular ALB appear?	In ICC-IF, HPA approves endoplasmic reticulum and Golgi localisation and lists Hep-G2 and U2OS images (HPA subcellular: approved locations and image cell lines). This intracellular observation does not replace the extracellular tissue IHC profile (HPA tissue IHC: profile).
Retrieval and species scope	No ALB-specific retrieval condition or nonhuman reactivity is established by these payloads. Optimize retrieval against controls and confirm species reactivity for the antibody being used (standard IHC practice); do not infer either from ALB topology (UniProt P02768: topology).

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
No signal at Bowman's capsule	The result differs from HPA's medium kidney observation; the cause is not identified by that observation (HPA tissue IHC: Kidney).	Check section morphology, the IHC-validated antibody's documented application, dilution and retrieval instructions, then verify detection with a positive control (standard IHC practice). No target-specific retrieval setting is supplied.
Whole slide has a brown haze	Broad background may obscure the reported plasma and extracellular matrix pattern (HPA tissue IHC: profile); incomplete blocking, washing or chromogen control are general possibilities (standard IHC practice).	Compare no-primary and detection controls, shorten development if overdeveloped, and adjust blocking or washing within the assay workflow (standard IHC practice). Preserve interpretable extracellular signal.
Strong signal appears in HPA-negative cells	Adipocytes and respiratory epithelial cells were not detected in the supplied observations (HPA tissue IHC: negative observations); nonspecific binding or endogenous detection activity may explain discordant color (standard IHC practice).	Inspect a matched negative section and no-primary control; verify detection blocking and repeat with an independently validated antibody if available (standard IHC practice).
Signal is mainly nuclear or outlines cell membranes	That distribution is inconsistent with secreted ALB lacking a transmembrane segment (UniProt P02768: location and topology), although the image alone cannot establish the cause.	Review focus, morphology, counterstain and negative controls; reassess antibody specificity before scoring the compartment as ALB (standard IHC practice).
Liver RNA enrichment is used to call an IHC section negative	RNA is liver-enriched, but HPA warns that RNA and protein tissue locations can differ for secreted variants (HPA tissue IHC: RNA specificity and reliability description).	Score the observed protein compartment against the plasma and extracellular matrix profile and include the documented kidney positive site where suitable (HPA tissue IHC: profile and Kidney).
An ICC-IF image seems to conflict with tissue IHC	HPA approves ER and Golgi localisation in ICC-IF, while its tissue IHC profile emphasizes plasma and extracellular matrix (HPA subcellular: approved locations; HPA tissue IHC: profile).	Interpret each assay in its own specimen context; for paraffin IHC, confirm the extracellular pattern and controls before calling a discrepancy (standard IHC practice).

Source: UniProt P02768 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing ALB staining

Immunohistochemistry Validation of Albumin in Human Liver Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-albumin antibody (A01245) at 2 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4 ˚C. A goat anti- chicken IgY (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

A01245 · IHCImmunohistochemistry Validation of Albumin in Human Liver Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-al

Immunohistochemistry Validation of Albumin in Mouse Liver Immunohistochemical analysis of paraffin-embedded mouse liver tissue using anti-albumin antibody (A01245) at 10 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4 ˚C. A goat anti- chicken IgY (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

A01245 · IHCImmunohistochemistry Validation of Albumin in Mouse Liver Immunohistochemical analysis of paraffin-embedded mouse liver tissue using anti-al

Immunofluorescence Validation of Albumin in Mouse Liver Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse liver tissue labeling albumin with A01245 at 20 ???????/mL, followed by goat anti-chicken IgY secondary antibody at 1/500 dilution (red) and DAPI staining (blue).

A01245 · IFImmunofluorescence Validation of Albumin in Mouse Liver Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse liver tissue labeling

M01245-4 · IHCHuman hepatocellular carcinoma was stained with anti-Albumin rabbit antibody

M01245-4 · IHCHuman kidney was stained with anti-Albumin rabbit antibody

M01245 · IHCImmunohistochemical analysis of paraffin-embedded human liver, using Serum Albumin Antibody.

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000163631-ALB/tissue/cerebral+cortex#imid_15793779

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000163631-ALB/tissue/colon#imid_15793705

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000163631-ALB/tissue/liver#imid_15793706

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000163631-ALB/tissue/kidney#imid_15793707

HPA · ICC-IF Hep-G2Human Protein Atlas immunofluorescence, Hep-G2, antibody HPA031024; green = ALB, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

Real IHC / IF data behind the staining-pattern expectations in this section.

Sample controls for ALB IHC & IF

🧪Run kidney first: Bowman's capsule must show staining (HPA: Medium in Bowman's capsule). Run adipose tissue as the negative, with adipocytes unstained (HPA: Not detected in adipocytes); on the kidney slide, assess neighboring cells outside Bowman's capsule for background without treating them as a validated negative population.

Positive control tissue: Kidney (Bowman's capsule, HPA Medium)

Negative control tissue: Adipose tissue (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ALB in Hep-G2, U2OS, with annotated localisation: Endoplasmic reticulum (approved), Golgi apparatus (approved) (HPA subcellular).

Technical controls: Include a no-primary (secondary-only) control, an isotype control matched to the primary antibody's host species and clonality, and a target-specific knockout sample or validated peptide-block control. For chromogenic kidney IHC, quench endogenous peroxidase and check for residual background (standard IHC practice).

⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A01245 tissue-IHC caption's fixative field is unreported (selected A01245 caption metadata). The caption uses heat retrieval in citrate buffer at pH 6 for paraffin-embedded liver, but does not establish whether kidney staining depends on that retrieval step (selected A01245 tissue-IHC caption). Neither frozen sections nor IF are established as easier by the supplied evidence; because ALB is secreted and occurs in plasma, assess extracellular or blood-associated staining separately from staining of the intended cells (UniProt P02768: secreted; tissue specificity: plasma).

HPA tissue IHC evidence for ALB

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — At least one protein variant secreted, tissue location of RNA and protein might differ and correlation is complex.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.

Positive expression · recommended positive controls

Tissue	Cell type	Level	Evidence	Source
Kidney	Bowman's capsule	Medium	Protein (IHC)	HPA →

Undetected expression · recommended 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 IHC Tips

Troubleshoot ALB staining by checking retrieval, compartment, tissue context, and detection controls before changing how the result is scored.

How should I troubleshoot weak ALB staining after antigen retrieval?

Use heat-mediated citrate buffer at pH 6 for ALB antigen retrieval in paraffin sections (datasheet A01245). The selected human liver IHC example used this retrieval method, followed by primary antibody incubation overnight at 4 °C (datasheet A01245). If staining is weak, check that sections reached and remained at the intended retrieval temperature, then compare a modest increase in heating time with the original run while monitoring tissue integrity (standard IHC practice). Assess the change against the same positive-control section and detection conditions, because a faint result alone cannot establish that retrieval is the limiting step (standard IHC practice).

Could fixation explain weak or uneven ALB staining?

ALB-specific sensitivity to fixation is unknown from the supplied evidence; do not infer it from tissue staining patterns or protein topology (supplied evidence; HPA tissue IHC; UniProt P02768). For paraffin-section IHC, compare sections with documented fixation histories and consistent thickness before changing antibody concentration (standard IHC practice). Excessive or uneven formaldehyde fixation can alter epitope access in IHC generally, so compare a controlled range of retrieval heating times while retaining citrate at pH 6 as the starting condition (standard IHC practice; datasheet A01245). If staining differs, report the fixation and processing histories alongside the result rather than assigning ALB-specific fixation sensitivity (standard IHC practice).

Where should convincing ALB staining appear in tissue?

Interpret extracellular and plasma-associated signal in light of ALB secretion and the HPA tissue profile of plasma and extracellular-matrix positivity (UniProt P02768: secreted; HPA tissue IHC). Intracellular signal can also be plausible in producing cells because the HPA subcellular record places ALB in the endoplasmic reticulum and Golgi apparatus (HPA subcellular). Liver is the enriched tissue for ALB RNA, whereas HPA reports medium staining in kidney Bowman's capsule (HPA tissue IHC). Score extracellular deposits separately from cellular staining, and inspect morphology and the matched negative control before treating diffuse luminal or vascular signal as evidence of local synthesis (standard IHC practice; UniProt P02768: secreted).

Can ALB processing or isoforms change what my antibody detects?

ALB has 3 reported isoforms, and its precursor contains a signal peptide at residues 1–18, a propeptide at 19–24, and an albumin chain at 25–609 (UniProt P02768). Therefore, confirm the catalog antibody's immunogen or mapped epitope before assuming that precursor and mature protein are recognized equally (UniProt P02768; standard IHC practice). The record also lists glycosylation and modified residues, but it does not establish that either changes staining with this antibody (UniProt P02768). If isoform discrimination matters, use an independently characterized epitope-specific reagent or orthogonal assay and describe ordinary chromogenic staining simply as ALB immunoreactivity (standard IHC practice).

How should I evaluate ALB in a multiplex IF experiment?

Treat IF/ICC as a separate application and verify its antibody performance directly; the selected A01245 evidence is paraffin-section chromogenic IHC (datasheet A01245). For liver multiplexing, pair ALB with a validated hepatocyte marker and check whether cellular signal matches the expected producing-cell population, while keeping extracellular signal distinct (HPA tissue RNA: liver enriched; UniProt P02768: secreted; standard IF practice). Choose a fluorophore and imaging channel that separate specific signal from measured tissue autofluorescence, with single-stain and no-primary controls (standard IF practice). For intracellular endoplasmic-reticulum or Golgi epitopes, assess fixation-compatible permeabilisation; for accessible extracellular ALB, assess staining without permeabilisation first (HPA subcellular; UniProt P02768: secreted; standard IF practice).

How can I reduce diffuse or vascular ALB background?

First distinguish expected plasma and extracellular-matrix ALB from nonspecific chromogen, since both compartments can stain in tissue (HPA tissue IHC). The selected liver example used 10% serum blocking for 1 hour at room temperature and primary antibody at 2 µg/ml overnight at 4 °C (datasheet A01245). If signal obscures structure, compare a lower primary concentration and more thorough washing while holding retrieval and exposure conditions constant (standard IHC practice). Include a no-primary control and an endogenous-peroxidase block when using HRP detection, then inspect whether residual color tracks vessels, edges, damaged tissue, or endogenous enzyme activity (standard IHC practice).

How should I quantify ALB staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION

Define the region and compartment before scoring because ALB is secreted and HPA reports plasma and extracellular-matrix positivity (UniProt P02768: secreted; HPA tissue IHC). For cellular staining, report the percentage of positive cells or an H-score using prespecified intensity bins; for extracellular deposits, measure positive area or optical density per mm² of viable tissue (standard IHC practice). Normalize each result to the relevant viable tissue area or eligible cell count, rather than total section area when empty spaces or necrosis vary (standard IHC practice). Keep section thickness, chromogen development, imaging settings, threshold, and control tissue consistent across comparisons, and state whether vascular signal was excluded (standard IHC practice).

When is ALB staining likely to be artefactual?

A convincing interpretation should fit ALB secretion, plasma-associated staining, and possible intracellular endoplasmic-reticulum or Golgi localisation in producing cells (UniProt P02768; HPA tissue IHC; HPA subcellular). Treat isolated strong staining in an unexpected cell population cautiously, particularly where extracellular protein could coat cells or occupy vessels (UniProt P02768: secreted; standard IHC practice). Edge-only color, necrotic areas, and staining persisting in a no-primary control warrant review for section or detection artefacts, including endogenous peroxidase with HRP methods (standard IHC practice). Compare intact morphology, the liver positive-control section, and matched controls before calling a weak or diffuse result true cellular ALB expression (HPA tissue RNA: liver enriched; standard IHC practice).

Boster reagents

Best ALB / Albumin IHC Antibodies

Anti-ALB IHC images cover human, mouse and rat liver, human hepatocellular carcinoma, and human and rat kidney (catalog IHC captions); an IF image covers mouse liver (A01245 IF caption).

Real IHC data

Anti-Albumin Antibody

Cat # A01245

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Real IHC data Human hepatocellular carcinoma was stained with anti-Albumin rabbit antibody

Anti-Albumin Rabbit Monoclonal Antibody

Cat # M01245-4

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Real IHC data Immunohistochemical analysis of paraffin-embedded human liver, using Serum Albumin Antibody.

Anti-Albumin Rabbit Monoclonal Antibody

Cat # M01245

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A01245 has IHC images from paraffin-embedded human, mouse and rat liver, plus an IF image from mouse liver (A01245 image captions). M01245-4 has IHC images from human hepatocellular carcinoma and human and rat kidney (M01245-4 IHC captions); M01245 has an IHC image from paraffin-embedded human liver (M01245 IHC caption).

Which to pick: For cross-species tissue IHC, choose A01245: its human, mouse and rat liver captions document paraffin sections, formaldehyde fixation and citrate pH 6 retrieval (A01245 IHC captions). For IF, A01245 has a mouse-liver image (A01245 IF caption); for ICC, A01245-4 lists the application but has no image in this payload (A01245-4 catalog applications and image captions). M01245-4 is a rabbit monoclonal with human and rat IHC images (M01245-4 catalog and IHC captions), while M01245 is a rabbit monoclonal shown on paraffin-embedded human liver (M01245 catalog and IHC caption); fixation is unreported in both products’ captions (M01245-4 and M01245 IHC captions).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

UniProt Consortium. UniProt entry P02768 (ALBU_HUMAN, Albumin).
Human Protein Atlas. ALB tissue IHC expression (reliability: Enhanced).
Human Protein Atlas. ALB subcellular location (ICC-IF): Localized to the Golgi apparatus and endoplasmic reticulum..
Human Protein Atlas. ALB antibody validation summary (3 antibodies).
Combined analysis of albumin in situ hybridisation and C reactive protein immunohistochemistry for the diagnosis of intrahepatic cholangiocarcinoma: towards a molecular classification paradigm. Journal of clinical pathology 2025 — PMC12015024.
Expression of Transferrin and Albumin in the Sperm-Storage Tubules of Japanese Quail and their Possible Involvement in Long-Term Sperm Storage. The journal of poultry science 2020 — PMC7063080.
Synergistic amelioration of cholestatic liver fibrosis by combined total astragalus saponins and AAV8.Numb-Exon3 through modulation of hepatic progenitor cell differentiation. Frontiers in pharmacology 2026 — PMC13149365.
Nrf2 induces malignant transformation of hepatic progenitor cells by inducing β-catenin expression. Redox biology 2022 — PMC9618468.
PubMed PMID:6171778 — UniProt-cited evidence.
PubMed PMID:6275391 — UniProt-cited evidence.
PubMed PMID:3009475 — UniProt-cited evidence.