AGBL2 / Cytosolic carboxypeptidase 2 · IHC design guide

Design Immunohistochemistry for AGBL2

Plan paraffin AGBL2 IHC with the IHC-validated antibody at 1:100–1:300 (datasheet) and assess cytoplasmic staining (HPA tissue IHC). Interpret tissue staining cautiously because agreement with RNA expression is very low (HPA tissue IHC).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
Immunohistochemistry protocol sheet for AGBL2 (IHC for AGBL2): expected localisation Cytoplasmic staining in tissue (HPA tissue IHC); centrosome and basal body localisation at the molecular level (UniProt), antibody A11275-1, validated IHC image, and IHC protocol steps
Printable AGBL2 IHC protocol sheet — expected localisation Cytoplasmic staining in tissue (HPA tissue IHC); centrosome and basal body localisation at the molecular level (UniProt), antibody A11275-1, controls and protocol steps. Open the full AGBL2 IHC guide →

AGBL2 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 Cytoplasmic staining in tissue (HPA tissue IHC); centrosome and basal body localisation at the molecular level (UniProt)
Staining pattern General cytoplasmic staining (HPA tissue IHC)
Antigen retrieval Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen)
Positive control ⓘ Adrenal gland+4 more · see all
Negative control ⓘ None in HPA (detected in all 44 tissues); use no-primary + isotype controls
Important caveats
Reasons your staining may differ from the expected pattern.
Fixation Keep fixation consistent across sections (standard IHC practice; not target-specific)
Caveat Very low staining–RNA consistency; verify specificity (HPA tissue IHC)
Regulation No specific expression regulator annotated (UniProt)
Isoform / epitope Three isoforms; epitope differences unreported (UniProt)
Section 1

Recommended AGBL2 IHC & IF Protocols

The catalog antibody’s IHC-P protocol is followed by published AGBL2 staining conditions for ovarian and renal tissues (PMC6781648; PMC7283229; PMC11417249).

Recommended immunohistochemistry (IHC-P) protocol parameters
SampleParaffin-embedded human heart tissue; fixative not specified (datasheet A11275-1)
FixationImage fixative and duration unreported (datasheet A11275-1); verify before use.
Sectioning4–5 µm sections on charged slides (standard)
DeparaffinisationXylene, graded ethanol series to water (standard)
Antigen retrievalHeat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen)
Peroxidase block3% H2O2, 10 min, room temperature (standard)
Blocking10% normal serum of the secondary host, 30 min, room temperature (standard)
Primary antibodyRabbit anti-AGBL2, 1:100-1:300 (datasheet A11275-1)
Primary incubationOvernight at 4 °C (standard)
DetectionHRP-polymer secondary, DAB chromogen 5–10 min (standard)
CounterstainHematoxylin, blue, dehydrate and mount (standard)
Expected resultAGBL2-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control.
💡Decision noteStart with citrate pH 6.0, 95–98 °C, 20 min (page antigen retrieval). If staining needs optimization, compare the published retrieval conditions (PMC6781648; PMC7283229; PMC11417249).
Section 2

What Is the Expected AGBL2 Staining Pattern?

AGBL2 is a cytosolic protein that also localizes to centrosomes, centrioles and ciliary basal bodies (UniProt Q5U5Z8 subcellular location; UniProt Q5U5Z8 topology). In paraffin sections, expect chiefly cytoplasmic staining in reported positive cells, including adrenal glandular cells and breast myoepithelial cells (HPA tissue IHC: High). Treat this as a provisional pattern: HPA rates tissue IHC reliability Uncertain because antibody staining and RNA expression have very low consistency (HPA tissue IHC: Uncertain).

What am I looking at on my slide?
Cytoplasmic staining in adrenal glandular cells or breast myoepithelial cells.This matches reported high staining in those cell types and the expected cytosolic compartment (HPA tissue IHC: High; UniProt Q5U5Z8 subcellular location). It is a plausible positive, but the HPA tissue pattern remains Uncertain; morphology alone does not establish antibody specificity (HPA tissue IHC: Uncertain).
Prominent staining confined to nuclei, cell membranes or extracellular material.This differs from the expected IHC cytoplasmic pattern and from UniProt’s nonmembrane, intracellular annotation (HPA tissue IHC: General cytoplasmic expression; UniProt Q5U5Z8 topology). Investigate background or cross-reactivity before scoring it as AGBL2. Nucleolar localization has separate ICC-IF support and should not be assumed to validate a nuclear IHC pattern (HPA subcellular: Nucleoli approved).
Strong signal in an unexpected cell population within the section.Compare the exact cell type with the HPA tissue record before assigning specificity (HPA tissue IHC: reported cell types; HPA tissue IHC: Uncertain). An unexpected chromogenic signal may reflect cross-reactivity or endogenous detection activity (standard IHC practice). HPA’s broad cytoplasmic profile does not define every unlisted cell type as negative (HPA tissue IHC: General cytoplasmic expression).
Diffuse colour across cells and surrounding tissue.A poorly bounded deposit is difficult to score as cellular AGBL2 staining (standard IHC practice). Check background controls, blocking and detection chemistry; widespread colour may arise from nonspecific binding or endogenous activity (standard IHC practice). Compare with the reported cellular, chiefly cytoplasmic pattern (HPA tissue IHC: General cytoplasmic expression).
No detectable signal in an HPA high-staining cell type.First confirm that the expected cells are present, then check section processing, retrieval, antibody dilution and detection controls (standard IHC practice). A negative result warrants investigation but cannot by itself disprove AGBL2 expression: the reported high pattern has Uncertain reliability and RNA discordance (HPA tissue IHC: High; HPA tissue IHC: Uncertain).
💡Expected AGBL2 appearanceCall a result provisionally positive when distinct cytoplasmic staining is visible in an HPA high-staining cell type; isolated membrane, extracellular or diffuse background colour is suspect (HPA tissue IHC: High; HPA tissue IHC: General cytoplasmic expression; UniProt Q5U5Z8 topology; standard IHC practice).
How each factor affects the staining
Tissue and cell choiceHPA reports high IHC staining in several cell types, including adrenal glandular, breast myoepithelial and bone marrow hematopoietic cells (HPA tissue IHC: High). Adipocytes are reported Low, not negative, so they are a limited contrast tissue (HPA tissue IHC: Low in adipocytes).
Evidence confidenceThe tissue staining profile is Uncertain because antibody staining and RNA data have very low consistency; HPA also flags a splice or transcript discrepancy and pending external verification (HPA tissue IHC: reliability description). HPA007718 is rated Uncertain for IHC (HPA antibodies: HPA007718 IHC Uncertain).
Intracellular distributionUniProt places AGBL2 in cytosol, centrioles and ciliary basal bodies, with no transmembrane segment (UniProt Q5U5Z8 subcellular location; UniProt Q5U5Z8 topology). Fine structures may be harder to resolve than broad cytoplasmic staining in routine chromogenic sections (standard IHC practice).
Isoforms and processingUniProt lists three isoforms and a single chain spanning residues 1–902, with no signal peptide or propeptide (UniProt Q5U5Z8 isoforms; UniProt Q5U5Z8 processing). Without an antibody epitope map, isoform coverage cannot be inferred; no supplied evidence supports a shed extracellular staining pattern (UniProt Q5U5Z8 processing; UniProt Q5U5Z8 topology).
IF/ICC Q&A: What localization should I expect?HPA ICC-IF places AGBL2 mainly in nucleoli, cytosol, basal bodies and flagellar centrioles; it also reports sperm mid, principal and end pieces (HPA subcellular). This informs localization interpretation only; HPA007718 has Uncertain ICC validation, and IF/ICC requires its own guide (HPA antibodies: HPA007718 ICC Uncertain).
Why is my staining missing, weak or wrong?
SituationLikely causeNext action
The expected cytoplasmic cells are unstained.The target cells may be absent from the section, or the staining run may have failed (standard IHC practice). The HPA positive call is itself Uncertain (HPA tissue IHC: Uncertain).Confirm cell identity on the counterstained section and review the positive control, retrieval, primary dilution and detection run (standard IHC practice). Repeat with an independently supported antibody if available (standard IHC practice).
Staining is mainly nuclear in paraffin sections.A nuclear-only IHC pattern conflicts with HPA’s general cytoplasmic tissue profile (HPA tissue IHC: General cytoplasmic expression). HPA’s approved nucleolar finding comes from ICC-IF (HPA subcellular: Nucleoli approved).Check localization against a matched control and another antibody before calling the nuclear IHC signal specific (standard IHC practice; HPA antibodies: HPA007718 IHC Uncertain).
A low-staining comparison tissue shows colour.Adipocytes are reported Low rather than negative; residual signal is possible (HPA tissue IHC: Low in adipocytes). Diffuse colour may also come from background detection activity (standard IHC practice).Assess the exact stained cell type and compare with a no-primary control; do not treat adipose tissue as an established AGBL2-negative control (HPA tissue IHC: Low in adipocytes; standard IHC practice).
Colour appears without primary antibody.The signal can arise from endogenous chromogenic detection activity or another reagent background source (standard IHC practice).Review the no-primary control, endogenous activity blocking and detection reagents, then interpret the test section only after background is controlled (standard IHC practice).
Signal covers tissue boundaries and obscures cells.Excess primary or detection signal, or insufficient blocking, can obscure cellular localization (standard IHC practice).Recheck primary dilution and blocking, reduce detection development as appropriate, and retain a matched control for comparison (standard IHC practice). Score only distinguishable cellular staining (standard IHC practice).
High staining disagrees with the expected RNA pattern.HPA reports very low consistency between antibody staining and RNA expression and flags a splice or transcript discrepancy (HPA tissue IHC: reliability description).Record the exact tissue, cell type and compartment; interpret the IHC result as provisional and seek independent antibody or orthogonal support before making an expression claim (HPA tissue IHC: Uncertain; standard IHC practice).

Sample controls for AGBL2 IHC & IF

🧪Run breast first and look for staining in myoepithelial cells (HPA: High in breast myoepithelial cells); use neighboring cells without signal as internal background references, without treating a named cell type as a proven negative. HPA detects AGBL2 in all 44 scored tissues, so no negative tissue is available; no-primary and isotype controls provide the negative baseline (HPA: no negative tissue rows).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: AGBL2 is detected in all 44 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show AGBL2 in ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), Sperm, with annotated localisation: Nucleoli (approved), Basal body (supported), Cytosol (approved), Flagellar centriole (supported) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) section and an isotype control matched to the primary antibody’s host and clonality (standard IHC practice); assess specificity with a knockout sample or peptide competition (A11275-1 tissue-IHC caption: peptide-blocked section). Quench endogenous peroxidase and check background in the breast section before interpreting weak chromogenic staining (standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the fixative for the A11275-1 paraffin-section caption is unreported (A11275-1 tissue-IHC caption). Antigen-retrieval dependency and whether frozen-section IHC or IF is easier are unreported in the supplied evidence. Endogenous peroxidase can create chromogenic background in breast tissue, so interpret staining against the control sections (standard IHC practice).

HPA tissue IHC evidence for AGBL2

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Very low consistency between antibody staining and RNA expression data. Pending external verification. Caution, Splice and/or transcript discrepancy exists.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.

Positive expression · recommended positive controls

TissueCell typeLevelEvidenceSource
Adrenal gland Glandular cells High Protein (IHC) HPA →
Appendix Glandular cells High Protein (IHC) HPA →
Bone marrow Hematopoietic cells High Protein (IHC) HPA →
Breast Myoepithelial cells High Protein (IHC) HPA →
Caudate Glial cells High Protein (IHC) HPA →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
None in HPA: AGBL2 is detected in all 44 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
Section 3

Advanced AGBL2 IHC Tips

Troubleshoot AGBL2 staining in paraffin sections by checking retrieval, cellular pattern, controls and scoring before interpreting signal as target specific.

How should I retrieve AGBL2 in paraffin sections when staining is weak?
Start with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval rule). Run a peptide-blocked section alongside the test section, since peptide blocking was shown for the catalog antibody in paraffin-embedded human heart tissue (A11275-1 tissue-IHC caption). If staining remains weak, compare a longer citrate retrieval with an alternative buffer in a small side-by-side trial, keeping detection exposure identical (standard IHC practice). Examine whether signal appears in cytoplasm or discrete centrosomal structures, which are compatible with reported AGBL2 locations (UniProt Q5U5Z8). Record section integrity and background as well as signal intensity, because excessive retrieval can damage morphology or increase nonspecific staining (standard IHC practice).
Could fixation explain weak or uneven AGBL2 staining?
Target-specific fixation sensitivity is unknown: the selected antibody image identifies paraffin-embedded human heart tissue but does not state its fixative (A11275-1 tissue-IHC caption). Document fixative, fixation duration, section thickness and processing history before comparing specimens (standard IHC practice). For a controlled comparison, process matched tissue pieces using a consistent routine fixation schedule and stain them in the same run (standard IHC practice). Check whether weak staining tracks with thick regions, folds or poorly preserved morphology before changing antibody conditions (standard IHC practice). Neither reported cytosolic localization nor the absence of annotated glycosylation establishes how fixation affects this epitope (UniProt Q5U5Z8).
Where should convincing AGBL2 staining appear in tissue sections?
Assess cytoplasmic staining first, because the tissue-IHC profile describes general cytoplasmic expression (HPA tissue IHC). Small structures near the nucleus or ciliary base may also be plausible: AGBL2 is reported at centrioles, centrosomes and basal bodies (UniProt Q5U5Z8). Use morphology and a matched control section to distinguish a discrete intracellular signal from diffuse chromogen deposited over tissue edges or folds (standard IHC practice). Strong nuclear-only staining deserves separate validation, although nucleolar localization has been reported in cell imaging (HPA subcellular). Avoid treating any one compartment as conclusive, since the tissue-IHC antibody evidence has uncertain reliability and low agreement with RNA data (HPA tissue IHC).
How can isoforms or epitope location affect AGBL2 IHC interpretation?
AGBL2 has 3 annotated isoforms, so an antibody's epitope determines which forms it can detect (UniProt Q5U5Z8; standard antibody practice). Confirm the catalog antibody's stated immunogen or epitope against the isoforms before interpreting absent staining as absent protein (standard antibody practice). The peptidase M14 domain spans residues 396–666, but that domain assignment alone does not locate this antibody's epitope (UniProt Q5U5Z8). Peptide blocking in the selected tissue image supports competition with the supplied peptide, without establishing isoform coverage or excluding every off-target reaction (A11275-1 tissue-IHC caption; standard antibody practice). Document the epitope and any splice-dependent uncertainty with the staining result, especially given the reported transcript discrepancy (HPA tissue IHC).
How can IF help assess an ambiguous chromogenic AGBL2 pattern?
Use IF as a separate corroboration experiment, while judging the paraffin-section IHC result on its own controls (standard IHC/IF practice). Multiplex AGBL2 with a marker for the cell type being evaluated, such as a validated glandular-cell marker when assessing reported glandular staining (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and put the weaker signal in a channel with low measured tissue autofluorescence (standard IF practice). Because AGBL2 lacks a transmembrane segment and is reported in the cytosol and basal body, permeabilize adequately for intracellular epitope access, then check morphology (UniProt Q5U5Z8; standard IF practice). HPA cell imaging reports cytosolic and basal-body localization, but those images do not validate this catalog antibody for IF (HPA subcellular; standard antibody practice).
What should I check when AGBL2 staining has high background?
Compare the test section with a no-primary control and the catalog antibody's peptide-blocked condition, recognizing that peptide competition alone cannot prove specificity (standard IHC practice; A11275-1 tissue-IHC caption). For peroxidase-based chromogenic detection, include an endogenous peroxidase block before applying the detection reagent and DAB (standard IHC practice). Optimize blocking and antibody concentration in a small titration while holding retrieval and development time constant; no target-specific dilution is supplied here (standard IHC practice). Check whether color accumulates at folds, cut edges, debris or damaged areas rather than within cells (standard IHC practice). Interpret faint diffuse cytoplasm cautiously because broad cytoplasmic staining is reported and tissue-IHC reliability is uncertain (HPA tissue IHC).
How should I score AGBL2 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then use the same thresholds across all sections (standard IHC practice). For cytoplasmic chromogen, report an H-score from intensity categories 0–3 weighted by the percentage of cells in each category, or report the percentage of positive cells alone (standard IHC practice). If focal structures are the outcome, count positive cells or structures per mm² of evaluable tissue rather than relying on whole-slide color density (standard image-analysis practice). Normalize comparisons to the number of eligible cells or evaluable tissue area, and exclude folds, necrosis and blank spaces consistently (standard IHC practice). State the compartment scored, because AGBL2 is reported in both cytosol and centrosome-related structures (UniProt Q5U5Z8).
How do I distinguish true AGBL2 staining from artefact?
Favor reproducible intracellular cytoplasmic or discrete centrosome-related staining over color confined to edges, folds or necrotic areas (UniProt Q5U5Z8; standard IHC practice). Compare the stained cell type with tissue morphology: reported examples include glandular cells in adrenal gland and appendix, while HPA rates the overall tissue-IHC reliability as uncertain (HPA tissue IHC). Investigate staining confined to an unexpected compartment or cell population with independent controls before assigning it to AGBL2 (standard IHC practice). A no-primary control helps reveal detection background, including endogenous enzyme activity in a peroxidase workflow (standard IHC practice). The selected peptide-blocked heart image provides a useful comparison, but peptide competition does not by itself establish target identity in every tissue (A11275-1 tissue-IHC caption; standard antibody practice).
Boster reagents

Best AGBL2 / Cytosolic carboxypeptidase 2 IHC Antibodies

Both anti-AGBL2 antibodies list IHC and IF for human and mouse (catalog: applications, reactivity). Each has a paraffin-section IHC image; neither has an IF image in the payload (catalog: image captions).

Real IHC data Immunohistochemistry analysis of paraffin-embedded human heart tissue, using CBCP2 Antibody. The picture on the right is blocked with the synthesized peptide.
Anti-CCP2 AGBL2 Antibody
Cat # A11275-1
Real IHC data Immunohistochemistry analysis of paraffin-embedded human heart tissue, using CBCP2 Antibody. The picture on the right is blocked with the synthesized peptide.
Anti-CBCP2 AGBL2 Antibody
Cat # A30633

A11275-1 lists IHC and IF for human and mouse, with an IHC image of paraffin-embedded human heart and a peptide-blocked comparison (catalog: A11275-1 applications, reactivity, image caption). A30633 lists IHC, IF and ICC for human and mouse, with its own IHC image of paraffin-embedded human heart and a peptide-blocked comparison (catalog: A30633 applications, reactivity, image caption).

Which to pick: For tissue IHC, either SKU has a paraffin-embedded human heart example (A11275-1 and A30633 image captions); the fixative is unreported in both captions. For IF/ICC, choose A30633 because it lists both applications and is described as polyclonal; A11275-1 lists IF but not ICC (catalog: applications, A30633 dilution data). For human or mouse samples, both list reactivity, but their IHC images document human heart only (catalog: reactivity; A11275-1 and A30633 image captions).

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

References

  1. UniProt Consortium. UniProt entry Q5U5Z8 (CBPC2_HUMAN, Cytosolic carboxypeptidase 2).
  2. Human Protein Atlas. AGBL2 tissue IHC expression (reliability: Uncertain).
  3. Human Protein Atlas. AGBL2 subcellular location (ICC-IF): Mainly localized to the nucleoli, cytosol, basal body and flagellar centriole. In addition localized to the mid piece, principal piece and end piece..
  4. Human Protein Atlas. AGBL2 antibody validation summary (1 antibodies).
  5. High expression of AGBL2 is a novel prognostic factor of adverse outcome in patients with ovarian carcinoma. Oncology letters 2019 — PMC6781648.
  6. Clinical implications of AGBL2 expression and its inhibitor latexin in breast cancer. World journal of surgical oncology 2014 — PMC4069086.
  7. Expression of RARRES1 and AGBL2 and progression of conventional renal cell carcinoma. British journal of cancer 2020 — PMC7283229.
  8. AGBL2 promotes renal cell carcinoma cells proliferation and migration via α-tubulin detyrosination. Heliyon 2024 — PMC11417249.
  9. PubMed PMID:14702039 — UniProt-cited evidence.
  10. PubMed PMID:16554811 — UniProt-cited evidence.
  11. PubMed PMID:15489334 — UniProt-cited evidence.