This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
ASRGL1 / Isoaspartyl peptidase/L-asparaginase · IHC design guide
Design Immunohistochemistry for ASRGL1
Plan chromogenic ASRGL1 IHC in paraffin sections using cytoplasmic staining in seminiferous ducts and female genital glands as a reference (HPA tissue IHC). Start the catalog antibody at 1:50 (datasheet A08869-1), and assess staining by cell type and compartment.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ASRGL1 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) | |
| Staining pattern | Cytoplasmic in seminiferous ducts and genital glands (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A08869-1) | |
| Positive control | Cerebellum+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A08869-1) | |
| Caveat | RNA may poorly predict staining: medium consistency (HPA tissue IHC) | |
| Regulation | Overexpressed in uterine and ovarian carcinomas (UniProt) | |
| Isoform / epitope | 2 isoforms; cleavage yields alpha and beta chains (UniProt) |
Section 1
Recommended ASRGL1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with published chromogenic ASRGL1 protocols for human brain and cervical tissue (PMC11099023; PMC6059725).
| Sample | Paraffin-embedded human brain tissue; fixative not specified (datasheet A08869-1) |
| Fixation | Image fixative and duration unreported (datasheet A08869-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A08869-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A08869-1) |
| Primary antibody | Rabbit anti-ASRGL1, 1:50 recommended; image 2 μg/ml (datasheet A08869-1) |
| Primary incubation | Overnight at 4 °C (datasheet A08869-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A08869-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ASRGL1-positive staining in granular cells - cytoplasm/membrane of cerebellum (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression most abundant in seminiferous ducts and glandular cells of female genital tract. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet A08869-1); citrate retrieval is a published alternative for cervical tissue (PMC6059725).
Section 2
What Is the Expected ASRGL1 Staining Pattern?
ASRGL1 is a cytoplasmic protein with no transmembrane segment (UniProt Q7L266). In paraffin section IHC, expect prominent staining in seminiferous ducts and female genital tract glandular cells (HPA: tissue IHC profile). HPA reports high staining in cervix and endometrium glandular cells, epididymis glandular cells, and cerebellar granular cells; its tissue IHC reliability is Enhanced, with medium consistency between staining and RNA data (HPA: tissue IHC).
| Distinct staining in cervix or endometrium glandular cells, or epididymis glandular cells. | This fits the reported High staining in those cell populations (HPA: tissue IHC). Judge the labelled cells and their cytoplasmic pattern, rather than calling an entire section positive from isolated deposits (HPA: tissue IHC profile; general IHC practice). |
| Predominantly nuclear staining in tissue IHC, with little cytoplasmic staining. | That differs from the reported tissue cytoplasmic profile (HPA: tissue IHC profile; UniProt Q7L266: cytoplasm). Check the counterstain and control sections before interpreting it as ASRGL1; nuclear localisation in ICC-IF does not establish a nuclear tissue IHC pattern (HPA: subcellular ICC-IF). |
| Strong signal in adipocytes or alveolar cells while the expected positive cells stain weakly. | HPA reports ASRGL1 as not detected in adipose tissue adipocytes and lung alveolar cells (HPA: tissue IHC). Treat this cell-specific mismatch as possible nonspecific binding or detection background; it does not establish that every cell in those tissues is negative (general IHC practice; HPA: tissue IHC). |
| Uniform colour over cells and surrounding section, without a readable cellular boundary. | Diffuse deposition cannot establish the cell-specific cytoplasmic pattern reported by HPA (HPA: tissue IHC profile). Review background controls and detection conditions before scoring, since nonspecific antibody binding or chromogen background can obscure localisation (general IHC practice). |
| No staining in glandular cells of a known-positive cervix or endometrium control. | Both populations have High reported staining (HPA: tissue IHC). First check whether the control section and detection run worked; absence in one section alone does not establish ASRGL1 absence in the specimen (general IHC practice). |
💡Expected ASRGL1 appearanceA convincing positive shows clear, predominantly cytoplasmic staining in HPA-reported High glandular cells or cerebellar granular cells; diffuse colour outside cell boundaries or dominant nuclear staining warrants investigation (HPA: tissue IHC; UniProt Q7L266: cytoplasm; general IHC practice).
| Cell-specific comparison tissue | Cervix and endometrium glandular cells, epididymis glandular cells, and cerebellar granular cells are reported High; bronchus basal cells and selected glial cells are Medium (HPA: tissue IHC). Compare like cell types when judging intensity. |
| Antibody validation | HPA lists IHC as Enhanced for HPA029725, HPA055572 and CAB070192 (HPA: antibodies). Its tissue profile still notes medium consistency with RNA data (HPA: tissue IHC); validation supports comparison with the reported pattern, not an assumption that every specimen stains. |
| Topology and compartment | ASRGL1 is cytoplasmic and has no transmembrane segment (UniProt Q7L266). Interpret tissue staining against the HPA cytoplasmic profile; cerebellar granular cells are specifically annotated as cytoplasm/membrane, so a membrane-associated edge there is not automatically discordant (HPA: tissue IHC). |
| Processing and isoforms | ASRGL1 autocleaves into alpha residues 1–167 and beta residues 168–308, and has two annotated isoforms (UniProt Q7L266). The supplied sources do not identify the antibodies’ epitopes, so they cannot establish which chain or isoform a stain detects. |
| IF/ICC Q&A: should its pattern match tissue IHC? | HPA reports mainly microtubules, with additional nucleoplasm and cytokinetic bridge localisation in ICC-IF (HPA: subcellular ICC-IF). Tissue IHC instead has a cytoplasmic profile (HPA: tissue IHC); interpret each application against its own reported pattern. |
| Situation | Likely cause | Next action |
|---|---|---|
| The positive control has no visible signal. | A failed staining or detection step is possible; high staining is reported in cervix and endometrium glandular cells (HPA: tissue IHC; general IHC practice). | Verify control tissue identity, antibody addition, retrieval, and detection reagents on the same run before scoring test sections (general IHC practice). |
| Only nuclei appear strongly labelled. | This conflicts with the reported tissue cytoplasmic profile, although HPA separately notes nucleoplasm in ICC-IF (HPA: tissue IHC; HPA: subcellular ICC-IF). | Compare with counterstain and negative controls, then review primary antibody concentration and detection background (general IHC practice). |
| Unexpected cells stain more strongly than the expected population. | Cell-specific cross-reactivity or detection background is possible when staining departs from the HPA cell map (HPA: tissue IHC; general IHC practice). | Identify the stained cell type, compare it with HPA’s cell-level entries, and inspect an appropriate negative control (HPA: tissue IHC; general IHC practice). |
| Colour is diffuse across tissue or outside cells. | Nonspecific binding, inadequate blocking, or excess chromogen development can obscure cellular staining (general IHC practice). | Review blocking, washes, antibody concentration, and development time; score only a resolved cellular pattern (general IHC practice). |
| Colour appears in a control without primary antibody. | Endogenous detection activity or detection reagent background may produce chromogenic signal independently of primary antibody (general IHC practice). | Check the detection controls and apply the appropriate endogenous activity block for the chosen detection system (general IHC practice). |
| A low-signal population appears negative. | HPA reports Low staining in several populations, including colon glandular cells and urinary bladder urothelial cells (HPA: tissue IHC). | Check a reported high-staining control in the same run before interpreting weak or absent staining in that population (HPA: tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing ASRGL1 staining
A08869-1 · IHCIHC analysis of ASRGL1 using anti-ASRGL1 antibody (A08869-1). ASRGL1 was detected in a paraffin-embedded section of human brain tissue. Heat
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000162174-ASRGL1/tissue/cerebral+cortex#imid_7501448
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000162174-ASRGL1/tissue/colon#imid_7497638
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000162174-ASRGL1/tissue/liver#imid_7497631
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000162174-ASRGL1/tissue/kidney#imid_7497640
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA029725; green = ASRGL1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA029725; green = ASRGL1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ASRGL1 IHC & IF
🧪Run cerebellum first and look for staining in granular cells (High; HPA: cerebellum, granular cells). Use adipose tissue as the negative tissue, scoring adipocytes as unstained (HPA: adipose tissue, adipocytes, Not detected). On the positive slide, use non-granular areas to assess background, but do not assume every other cell type is ASRGL1-negative (HPA: cerebellum, granular cells).
Positive control tissue: Cerebellum (Granular cells - cytoplasm/membrane, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ASRGL1 in A-431, U-251MG, U2OS, with annotated localisation: Microtubules (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit IgG isotype control (selected-SKU caption: rabbit primary antibody); use ASRGL1 knockout tissue or cells, if available, as a biological negative (standard IHC practice). Quench endogenous peroxidase before HRP/DAB detection in cerebellum (selected-SKU caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected-SKU brain caption does not state the fixative (selected-SKU caption: fixative not stated). The documented paraffin-section workflow used heat retrieval in EDTA, pH 8.0; that supports trying this retrieval condition but does not establish that retrieval is required for every specimen (selected-SKU caption: brain IHC). Frozen sections and IF are not established as easier by the supplied evidence; brain autofluorescence may complicate IF interpretation (standard IF practice).
HPA tissue IHC evidence for ASRGL1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data.). 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 |
|---|---|---|---|---|
| Cerebellum | Granular cells - cytoplasm/membrane | High | Protein (IHC) | HPA → |
| Cervix | Glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | High | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Basal cells | 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 → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ASRGL1 IHC Tips
Troubleshoot ASRGL1 staining in paraffin sections by checking retrieval, cellular pattern, controls, and scoring before interpreting signal intensity.
Which retrieval conditions should I try first for ASRGL1 staining?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A08869-1). The selected brain-section image used that buffer, followed by 10% goat serum blocking and 2 μg/ml primary antibody overnight at 4°C (datasheet A08869-1). If staining is weak, adjust heating time in small steps while keeping buffer and detection conditions constant, and compare with an untreated section (standard IHC practice). Review tissue preservation and staining of a known positive region before changing retrieval chemistry (standard IHC practice; HPA: high staining in cerebellar granular cells). Excessive retrieval can damage morphology and make compartment-level interpretation unreliable (standard IHC practice).
Could fixation explain weak or patchy ASRGL1 staining?
The selected paraffin-section caption does not state a fixative, so ASRGL1 sensitivity to any particular fixation condition is unknown for this antibody (datasheet A08869-1). Record the fixative, fixation duration, section age, and retrieval history for each specimen, then compare matched sections processed together (standard IHC practice). Use EDTA at pH 8.0 as the retrieval starting point and keep the 2 μg/ml primary incubation constant while assessing processing differences (datasheet A08869-1). Include a well-preserved positive-control section so widespread signal loss can be distinguished from regional biology (standard IHC practice; HPA: high staining in cerebellar granular cells). Do not infer a fixation effect from tissue-expression patterns or protein topology alone (HPA: tissue IHC profile; UniProt Q7L266 topology).
How should I assess cytoplasmic, membranous, or nuclear ASRGL1 staining?
Assess staining in identified cells and record cytoplasmic, membrane-associated, and nuclear signal separately (standard IHC practice). UniProt places ASRGL1 in the cytoplasm and reports no transmembrane segment, while HPA describes cytoplasmic or membrane staining in cerebellar granular cells (UniProt Q7L266 subcellular location and topology; HPA: cerebellum). HPA cell imaging also places ASRGL1 mainly on microtubules, with additional nucleoplasmic and cytokinetic-bridge signal; that imaging pattern should not be treated as a required appearance in chromogenic sections (HPA: subcellular). If staining appears exclusively as a crisp cell-surface rim, compare it with the expected cytoplasmic signal in a positive section and check controls (UniProt Q7L266 subcellular location; standard IHC practice). Avoid scoring a compartment without first confirming that the stained structures are intact cells (standard IHC practice).
Can processing or isoforms change what this antibody detects in sections?
ASRGL1 has 2 annotated isoforms and undergoes autocleavage into an alpha chain spanning residues 1–167 and a beta chain spanning 168–308 (UniProt Q7L266 isoforms and processing). The supplied caption does not identify this antibody’s epitope, so its recognition of each isoform or processed chain cannot be assigned from the staining image (datasheet A08869-1). Request an epitope map or compare independently validated reagents before interpreting a missing compartment as isoform-specific expression (standard IHC practice). ASRGL1 has no annotated transmembrane segment or glycosylation sites; neither feature establishes antibody accessibility after paraffin processing (UniProt Q7L266 topology and glycosylation; standard IHC practice). Keep retrieval and detection matched when comparing putative isoform-associated patterns across sections (standard IHC practice).
How can I follow up an IHC pattern with multiplex IF?
Use IF as a separate validation experiment: the selected antibody evidence documents chromogenic staining of a paraffin brain section, without specifying its fixative or an IF protocol (datasheet A08869-1). Multiplex ASRGL1 with a separately validated marker for the cell type being assessed, such as the granular-cell population identified in cerebellum, and include single-label controls (HPA: high staining in cerebellar granular cells; standard IF practice). Choose spectrally separated fluorophores and inspect unstained tissue for autofluorescence before assigning a dim channel (standard IF practice). Because ASRGL1 lacks a transmembrane segment and is reported in the cytoplasm, test permeabilisation for intracellular access, adjusting it to preserve cell structure (UniProt Q7L266 topology and subcellular location; standard IF practice). Do not transfer the IHC retrieval conditions to IF without testing them (datasheet A08869-1; standard IF practice).
What should I check when DAB background obscures ASRGL1 staining?
Run a no-primary control and inspect blank areas, section edges, and damaged tissue before changing the primary concentration (standard IHC practice). The selected section used 10% goat serum, 2 μg/ml rabbit primary overnight at 4°C, and a peroxidase-conjugated secondary for 30 minutes at 37°C (datasheet A08869-1). A peroxidase block and careful DAB development are general chromogenic IHC controls, rather than ASRGL1-specific validation steps (standard IHC practice). If background persists, titrate primary antibody and detection time against a positive section while preserving EDTA retrieval at pH 8.0 for the initial comparison (datasheet A08869-1; standard IHC practice). Score only cellular signal that exceeds the matched control (standard IHC practice).
How should I quantify heterogeneous ASRGL1 staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then use an H-score, percentage of positive cells, or positive-cell density per mm² consistently across specimens (standard IHC practice). An H-score combines the percentage of cells at each intensity with an ordinal intensity grade; record both inputs so regional differences remain auditable (standard IHC practice). Normalize counts to the number of evaluable cells, or density to the measured viable tissue area, and exclude folds and necrosis by a prespecified rule (standard IHC practice). Analyze comparable anatomical regions because HPA reports high staining in cerebellar granular cells but low staining in cerebral-cortex glial cells (HPA: cerebellum and cerebral cortex). Keep retrieval, antibody incubation, DAB development, and image settings matched across the comparison (datasheet A08869-1; standard IHC practice).
What distinguishes credible ASRGL1 staining from an artefact?
Look for reproducible signal in intact cells with a predominantly cytoplasmic pattern, interpreted against local anatomy and a matched control (UniProt Q7L266 subcellular location; standard IHC practice). High staining in cerebellar granular cells and glandular cells of cervix or endometrium provides tissue-level context; these patterns do not prove specificity in an individual section (HPA: cerebellum, cervix, and endometrium; standard IHC practice). Treat isolated edge staining, necrotic deposits, or signal reproduced in a no-primary control as suspect (standard IHC practice). Evaluate strong surface-only staining cautiously because ASRGL1 has no transmembrane segment, while allowing for HPA’s reported cytoplasm/membrane pattern in cerebellar granular cells (UniProt Q7L266 topology; HPA: cerebellum). Exclude endogenous peroxidase activity with an appropriate control before attributing DAB deposits to ASRGL1 (standard IHC practice).
Boster reagents
Best ASRGL1 / Isoaspartyl peptidase/L-asparaginase IHC Antibodies
A08869-1 has an IHC image from a paraffin-embedded human brain section (A08869-1 IHC caption); human, mouse and rat reactivity is listed (catalog: reactivity).
Real IHC data
Anti-ASRGL1 Antibody ®
Cat # A08869-1
A08869-1 is listed for IHC and for human, mouse and rat reactivity (catalog: applications/reactivity). Its own image documents chromogenic IHC on paraffin-embedded human brain with EDTA retrieval at pH 8.0 and DAB detection; the fixative is unreported (A08869-1 IHC caption).
Which to pick: For tissue IHC, choose A08869-1: this rabbit polyclonal antibody has an IHC application listing and an image from a paraffin-embedded human brain section (catalog: host/clonality/applications; A08869-1 IHC caption). For IF/ICC, A08869-1 has a listed 1:50 dilution, but no IF/ICC application listing or image; validate its use in your sample (catalog: if_dilution/applications/if_image_alts). For mouse or rat tissue, A08869-1 lists reactivity with both species, while its IHC image documents human tissue only (catalog: reactivity; A08869-1 IHC caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q7L266 (ASGL1_HUMAN, Isoaspartyl peptidase/L-asparaginase).
- Human Protein Atlas. ASRGL1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. ASRGL1 subcellular location (ICC-IF): Mainly localized to the microtubules. In addition localized to the nucleoplasm and cytokinetic bridge..
- Human Protein Atlas. ASRGL1 antibody validation summary (3 antibodies).
- TDP-43 proteinopathy in ALS is triggered by loss of ASRGL1 and associated with HML-2 expression. Nature communications 2024 — PMC11099023.
- Deletion of Asrgl1 Leads to Photoreceptor Degeneration in Mice. Frontiers in cell and developmental biology 2021 — PMC8805730.
- RNAi‑mediated downregulation of asparaginase‑like protein 1 inhibits growth and promotes apoptosis of human cervical cancer line SiHa. Molecular medicine reports 2018 — PMC6059725.
- A Mouse Model with Ablated Asparaginase and Isoaspartyl Peptidase 1 (Asrgl1) Develops Early Onset Retinal Degeneration (RD) Recapitulating the Human Phenotype. Genes 2022 — PMC9408336.
- PubMed PMID:11984834 — UniProt-cited evidence.
- PubMed PMID:14654938 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ASRGL1 (Isoaspartyl peptidase/L-asparaginase). Expected localisation Cytoplasmic staining in tissue (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A08869-1). Validated IHC antibody: A08869-1 at 1:50 recommended; image 2 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (Granular cells - cytoplasm/membrane); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human brain tissue; fixative not specified (datasheet A08869-1); 2. Fixation: Image fixative and duration unreported (datasheet A08869-1); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A08869-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A08869-1); 8. Primary antibody: Rabbit anti-ASRGL1, 1:50 recommended; image 2 μg/ml (datasheet A08869-1); 9. Primary incubation: Overnight at 4 °C (datasheet A08869-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A08869-1); Caveat: RNA may poorly predict staining: medium consistency (HPA tissue IHC); Isoform / epitope: 2 isoforms; cleavage yields alpha and beta chains (UniProt); Regulation: Overexpressed in uterine and ovarian carcinomas (UniProt). Evidence: UniProt Q7L266, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the microtubules. In addition localized to the…, Datasheet A08869-1, PMC11099023, PMC6059725. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ASRGL1/immunohistochemistry-ihc-guide.