ADPGK / ADP-dependent glucokinase · IHC design guide

Design Immunohistochemistry for ADPGK

Plan ADPGK paraffin IHC around the observed cytoplasmic tissue pattern (HPA tissue IHC). Colon glandular cells and lung macrophages provide high-staining references (HPA tissue IHC); start the catalog antibody at 1:100–1:300 (datasheet A10813-1).

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

Immunohistochemistry protocol sheet for ADPGK (IHC for ADPGK): expected localisation Cytoplasmic tissue staining (HPA tissue IHC), antibody A10813-1, validated IHC image, and IHC protocol steps — Printable **ADPGK** IHC protocol sheet — expected localisation Cytoplasmic tissue staining (HPA tissue IHC), antibody A10813-1, controls and protocol steps. Open the full ADPGK IHC guide →

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ADPGK 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 tissue staining (HPA tissue IHC)
	Staining pattern	Widespread cytoplasmic staining across cell types (HPA tissue IHC)
	Antigen retrieval	Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A10813-1)
	Positive control ⓘ	Colon+4 more · see all
	Negative control ⓘ	Bone marrow+4 more · see all

Important caveats

Reasons your staining may differ from the expected pattern.

	Fixation	Keep fixation consistent across samples (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 A10813-1)
	Caveat	Secreted annotation may complicate cytoplasmic scoring (UniProt)
	Regulation	Specific staining regulators are unreported (UniProt)
	Isoform / epitope	6 isoforms; signal peptide 1–22 is removed; epitope unknown (UniProt)

Section 1

Recommended ADPGK IHC & IF Protocols

Compare the catalog antibody’s IHC-P protocol (datasheet A10813-1) with four published ADPGK IHC protocols (PMC11974013; PMC13266032; PMC7532211; PMC10714548).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A10813-1)
Fixation	Image fixative and duration unreported (datasheet A10813-1); verify before use.
Sectioning	4–5 µm sections on charged slides (standard)
Deparaffinisation	Xylene, graded ethanol series to water (standard)
Antigen retrieval	Heat retrieval: Tris-EDTA pH 9.0 (datasheet A10813-1); 20 min, 95–100 °C (standard)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% normal serum of the secondary host, 30 min, room temperature (standard)
Primary antibody	Rabbit anti-ADPGK, 1:100-1:300 (datasheet A10813-1)
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	ADPGK-positive staining in glandular cells of colon (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression. No signal in the no-primary control.

💡Decision noteStart with heat-mediated Tris-EDTA pH 9.0 retrieval (datasheet A10813-1); the published excerpts do not specify retrieval conditions (PMC11974013; PMC13266032; PMC7532211; PMC10714548).

Section 2

What Is the Expected ADPGK Staining Pattern?

For paraffin-section IHC, expect predominantly cytoplasmic ADPGK staining across many cell types, with high staining in colon, rectum and small-intestinal glandular cells and lung macrophages (HPA: ubiquitous cytoplasmic expression; High in these cells). HPA rates its tissue IHC evidence Approved, with medium consistency against RNA expression (HPA: reliability summary). UniProt annotates ADPGK as secreted with no transmembrane segment, so interpret the observed cytoplasmic pattern in that context (UniProt Q9BRR6: subcellular location and topology).

What am I looking at on my slide?
Cytoplasmic chromogen in glandular cells of colon, rectum or small intestine, or in lung macrophages.	These are useful positive-pattern checks: HPA reports High staining in each named cell population and describes the overall tissue profile as ubiquitous cytoplasmic expression (HPA: tissue IHC). Assess the named cells, since an impression of staining across the whole section does not establish which cell population is positive (standard IHC practice).
Predominantly nuclear or membrane-restricted signal, with little cytoplasmic staining in an expected positive cell population.	That compartment pattern diverges from the reported tissue IHC profile (HPA: ubiquitous cytoplasmic expression). UniProt reports no transmembrane segment but annotates the protein as secreted (UniProt Q9BRR6: topology; subcellular location). Treat the discrepancy as a reason to check staining specificity and slide controls, not as proof of a new ADPGK location (standard IHC practice).
Strong stain in bronchial respiratory epithelium, esophageal or oral squamous epithelium, or smooth muscle cells.	HPA lists ADPGK as Not detected in those specific cell populations (HPA: tissue IHC). Check whether the color follows the intended cellular compartment and persists with appropriate detection controls; antibody cross-reactivity or endogenous detection activity can mimic a positive result (standard IHC practice). A negative HPA entry does not make every cell in that tissue negative.
Widespread haze or chromogen over cells and surrounding tissue, obscuring cellular borders.	Diffuse background cannot establish ADPGK localization or a High-versus-Medium cellular pattern (standard IHC practice; HPA: tissue IHC levels). Inspect the no-primary control, blocking and wash performance, and compare a known positive cell population on the same run before assigning biological meaning (standard IHC practice).
No discernible cytoplasmic staining in the named cells of a colon, rectum or small-intestinal section.	That result conflicts with HPA's High glandular-cell staining in those tissues (HPA: tissue IHC). It warrants a run-level check of positive control, retrieval and detection before being interpreted as absence of ADPGK (standard IHC practice). HPA's Approved rating has medium RNA–staining consistency, so one image profile is a reference rather than an absolute cutoff (HPA: reliability summary).

💡Expected ADPGK appearanceCall a positive IHC result when cytoplasmic staining is evident in glandular cells of colon, rectum or small intestine, or lung macrophages, at a level consistent with HPA's High examples; isolated nuclear, membrane-only or diffuse noncellular color is a specificity concern rather than the reported tissue pattern (HPA: tissue IHC; standard IHC practice).

How each factor affects the staining
Which observations anchor the tissue comparison?	HPA reports High staining in the named intestinal glandular cells and lung macrophages, Medium staining in several other glandular populations, and Low or Not detected staining in specified cells elsewhere (HPA: tissue IHC). Compare cell type as well as tissue name.
How strong is the IHC pattern evidence?	The tissue profile is Approved but has medium consistency between antibody staining and RNA expression; the listed IHC antibody HPA045194 is Approved, while HPA058525 is listed for ICC rather than IHC (HPA: reliability summary; antibody validation).
Does protein processing predict a different slide compartment?	UniProt annotates a signal peptide at residues 1–22, a chain at 23–497, secretion and no transmembrane segment (UniProt Q9BRR6: processing; topology; location). These annotations do not replace the cytoplasmic tissue IHC observation (HPA: tissue IHC).
Do isoforms explain a discrepant stain?	UniProt lists 6 isoforms (UniProt Q9BRR6: isoforms). The supplied record gives no antibody epitope or isoform-specific IHC pattern, so an unexpected positive or negative result cannot be assigned to a particular isoform from these sources.
What should an IF/ICC result look like?	On its separate guide page, compare IF/ICC images with the approved centrosome localization shown in SiHa, U-251MG and U2OS cells (HPA: subcellular ICC-IF). That ICC-IF observation answers the localization question; it does not redefine the cytoplasmic paraffin-section IHC reference (HPA: tissue IHC).

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Expected positive glandular cells are blank.	The result disagrees with HPA's High colon, rectum and small-intestinal glandular-cell staining; the supplied sources do not identify a target-specific fixation sensitivity (HPA: tissue IHC).	Verify the section contains the named cells and that a positive control stains; then check the established retrieval, primary-antibody dilution and detection steps for this IHC run (standard IHC practice).
Nuclei dominate while cytoplasm is weak.	Nuclear-dominant staining differs from the reported ubiquitous cytoplasmic IHC pattern (HPA: tissue IHC). The pattern alone cannot identify whether the cause is nonspecific binding or a preparation issue (standard IHC practice).	Compare the no-primary control and a known positive tissue, inspect localization at cellular resolution, and confirm that the IHC-validated antibody was used (standard IHC practice; HPA: HPA045194 IHC Approved).
Bronchial respiratory epithelium or smooth muscle appears strongly positive.	Those named cell populations are listed as Not detected, so strong staining requires a specificity check (HPA: tissue IHC). Endogenous detection activity and antibody cross-reactivity are possible explanations (standard IHC practice).	Review the no-primary and detection controls, then compare with a High positive population processed in the same run; score each cell type separately (standard IHC practice; HPA: tissue IHC).
Brown color is diffuse across the section.	Broad noncellular deposit prevents reliable assignment of the cytoplasmic pattern reported by HPA (HPA: tissue IHC; standard IHC practice). Inadequate blocking, washing or detection control can contribute to background (standard IHC practice).	Inspect the no-primary control and reagent background; repeat the established blocking and wash steps and assess the signal only where cell boundaries are clear (standard IHC practice).
A purported negative tissue contains scattered positive cells.	HPA's Not detected entries refer to specified populations, such as bone-marrow hematopoietic cells or bronchial respiratory epithelial cells, rather than every cell in a section (HPA: tissue IHC).	Identify the stained cell population before scoring, compare its compartment with HPA's cytoplasmic IHC profile, and use controls to evaluate unexpected staining (HPA: tissue IHC; standard IHC practice).
The IHC result is used to assess an IF/ICC image.	HPA reports cytoplasmic tissue IHC and approved centrosome localization by ICC-IF; these are distinct assay observations (HPA: tissue IHC; subcellular ICC-IF).	Evaluate paraffin-section IHC against the named tissue cell populations, and evaluate IF/ICC localization against the separate HPA centrosome observation (HPA: tissue IHC; subcellular ICC-IF).

Source: UniProt Q9BRR6 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing ADPGK staining

Immunohistochemical analysis of paraffin-embedded human tonsil. 1, Antibody was diluted at 1:200 (4° overnight). 2, Tris-EDTA, pH9.0 was used for antigen retrieval. 3, Secondary antibody was diluted at 1:200 (room temperature, 30min).

A10813-1 · IHCImmunohistochemical analysis of paraffin-embedded human tonsil. 1, Antibody was diluted at 1:200 (4° overnight). 2, Tris-EDTA, pH9.0 was use

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000159322-ADPGK/tissue/cerebral+cortex#imid_12867575

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000159322-ADPGK/tissue/colon#imid_12866117

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000159322-ADPGK/tissue/liver#imid_12866126

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000159322-ADPGK/tissue/kidney#imid_12866119

HPA · ICC-IF SiHaHuman Protein Atlas immunofluorescence, SiHa, antibody HPA058525; green = ADPGK, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

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

Sample controls for ADPGK IHC & IF

🧪Run colon first and look for staining in glandular cells (HPA: High in colon glandular cells). Use bone marrow hematopoietic cells as the negative tissue (HPA: Not detected in bone marrow hematopoietic cells); on the colon slide, neighboring cells that remain unstained should show counterstain without specific chromogen, serving as an internal background comparison.

Positive control tissue: Colon (Glandular cells, HPA High)

Negative control tissue: Bone marrow (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ADPGK in SiHa, U-251MG, U2OS, with annotated localisation: Centrosome (approved) (HPA subcellular).

Technical controls: Include a no-primary, secondary-only control and a nonimmune antibody control matched to the primary antibody’s host species and immunoglobulin class; match the isotype for a monoclonal primary. Confirm specificity with knockout tissue or cells, or a peptide block if the immunizing peptide is available; for chromogenic colon IHC, quench endogenous peroxidase and inspect glandular and luminal background (HPA: High in colon glandular cells).

⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A10813-1 tonsil caption does not state a fixative. For paraffin IHC, its Tris-EDTA retrieval at pH 9.0 and 1:200 primary dilution at 4°C overnight provide starting conditions (A10813-1 tissue-IHC caption); review colon glandular staining against luminal background (HPA: High in colon glandular cells). The supplied evidence does not establish that frozen sections or IF are easier; IF localisation should be assessed independently of tissue IHC (HPA: centrosome localisation in ICC-IF).

HPA tissue IHC evidence for ADPGK

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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
Colon	Glandular cells	High	Protein (IHC)	HPA →
Lung	Macrophages	High	Protein (IHC)	HPA →
Rectum	Glandular cells	High	Protein (IHC)	HPA →
Small intestine	Glandular cells	High	Protein (IHC)	HPA →
Adrenal gland	Glandular cells	Medium	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Bone marrow	Hematopoietic cells	Not detected	Protein (IHC)	HPA →
Bronchus	Respiratory epithelial cells	Not detected	Protein (IHC)	HPA →
Caudate	Glial cells	Not detected	Protein (IHC)	HPA →
Cerebellum	Cells in granular layer	Not detected	Protein (IHC)	HPA →
Esophagus	Squamous epithelial cells	Not detected	Protein (IHC)	HPA →

Section 3

Advanced ADPGK IHC Tips

Troubleshoot ADPGK staining in paraffin sections by checking retrieval, cell identity and compartment before comparing chromogenic signal across samples.

What should I change when ADPGK staining is weak after antigen retrieval?

Use heat-mediated Tris-EDTA at pH 9.0 for antigen retrieval of paraffin sections (datasheet A10813-1). The selected ADPGK tissue image used that buffer with antibody at 1:200 overnight at 4°C (caption A10813-1). If staining remains weak, compare retrieval heating and cooling conditions on adjacent sections while holding antibody incubation and detection constant (standard IHC practice). Include a positive control such as colon glandular cells, where staining is reported as high (HPA: High in colon glandular cells). Evaluate tissue preservation and background alongside signal, since excessive heating can damage morphology and make apparent gains difficult to interpret (standard IHC practice).

Can I optimize fixation specifically for ADPGK using the available tissue image?

Target-specific ADPGK sensitivity to fixation is unknown from the supplied evidence; the selected paraffin-section caption does not state a fixative (caption A10813-1). Record the fixative, fixation interval and processing conditions for each specimen before comparing chromogenic staining (standard IHC practice). When weak staining tracks with a processing batch, compare similarly sampled sections processed under documented conditions while keeping Tris-EDTA retrieval at pH 9.0 constant (datasheet A10813-1; standard IHC practice). Review nuclear detail, tissue integrity and staining of a positive control in the same run (standard IHC practice). Neither reported tissue staining nor ADPGK sequence features establish a fixation effect (HPA: tissue IHC; UniProt Q9BRR6).

How should I judge diffuse cytoplasmic versus punctate ADPGK staining?

Assess chromogenic tissue staining first against the reported ubiquitous cytoplasmic profile (HPA: tissue IHC). The separate cell-imaging record lists the centrosome as an approved location, so a small punctum can be plausible without making every tissue punctum specific (HPA: subcellular). UniProt annotates ADPGK as secreted, with a 1–22 signal peptide and no transmembrane segment (UniProt Q9BRR6). Score cytoplasmic staining by cell type and compare puncta with tissue architecture, counterstain and a negative control (standard IHC practice). Do not assign extracellular deposits or isolated nuclear staining to ADPGK solely from color; check whether the pattern survives changes in detection background (standard IHC practice; HPA: tissue IHC).

Could ADPGK isoforms or epitope placement explain inconsistent tissue staining?

ADPGK has 6 annotated isoforms, but the supplied evidence does not map the catalog antibody epitope to them (UniProt Q9BRR6; caption A10813-1). Its ADPK domain spans residues 52–497, while the annotated signal peptide spans 1–22 (UniProt Q9BRR6). Ask for the antibody's immunogen or epitope coordinates before claiming that a staining difference reflects a particular splice form (standard antibody-validation practice). Compare serial sections with the same retrieval, dilution and detection settings to determine whether the discrepancy follows specimen processing (standard IHC practice). If an independently validated antibody recognizing another epitope is available, concordant cell patterns can strengthen the interpretation, while disagreement needs further validation (standard IHC practice).

How can IF help resolve an ambiguous ADPGK IHC pattern?

Use IF/ICC as a separate localisation check, with multiplex staining for a marker of the cell population under investigation (standard IF practice). For example, lung macrophages are reported as ADPGK-high, so a validated macrophage marker can test whether their signal overlaps (HPA: High in lung macrophages; standard IF practice). Choose fluorophores whose emission can be distinguished from the specimen's autofluorescence, and inspect single-channel and secondary-only controls (standard IF practice). Match permeabilisation to the mapped epitope's membrane accessibility: intracellular epitopes require access, whereas extracellular-accessible epitopes can be assessed without it (standard IF practice). Interpret cytoplasmic signal and possible centrosomal puncta against the distinct tissue and cell-imaging records (HPA: tissue IHC; HPA: subcellular).

How do I reduce widespread brown staining without losing ADPGK signal?

Run a no-primary control beside a positive section to locate staining from the detection system (standard IHC practice). Block endogenous peroxidase before peroxidase-based chromogenic detection and compare the DAB development interval across matched sections (standard IHC practice). Titrate the primary around the documented 1:200 overnight condition, changing one variable at a time (caption A10813-1; standard IHC practice). Examine whether color follows edges, folds or damaged areas rather than intact cell cytoplasm, and improve washing or blocking if that pattern persists (standard IHC practice; HPA: tissue IHC). Retain a cell-type control such as colon glandular cells to check that a background reduction has preserved detectable ADPGK staining (HPA: High in colon glandular cells).

What should I measure when comparing ADPGK staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION

Define the tissue compartment and eligible cells before scoring, then keep those rules fixed across slides (standard IHC practice). For cytoplasmic staining, report the percentage of positive cells and an H-score from 0–300 using intensity categories 0–3 (standard IHC practice; HPA: tissue IHC). If positive cells are sparse, report their density per mm² of viable tissue and state the sampled area (standard IHC practice). Normalise counts to the relevant cell population or viable tissue area, rather than total section area when tissue content differs (standard IHC practice). Score cell types separately because reported ADPGK levels vary between glandular cells, macrophages and other populations (HPA: tissue IHC).

When is an apparent ADPGK-positive IHC result likely to be artefactual?

Give greatest weight to staining in intact cytoplasm and in identified cells, consistent with the reported tissue profile (HPA: tissue IHC; standard IHC practice). High staining is reported in lung macrophages and colon glandular cells, whereas bronchial respiratory epithelial cells are listed as not detected (HPA: tissue IHC). Treat isolated nuclear color, section-edge enhancement and staining restricted to necrotic material as patterns requiring additional controls (standard IHC practice; HPA: tissue IHC). A no-primary section can reveal endogenous enzyme or detection-related color, while a matched positive section checks assay performance (standard IHC practice). Reconcile any discrete puncta with the approved centrosome localisation from cell imaging, without assuming that every tissue punctum represents ADPGK (HPA: subcellular; standard IHC practice).

Boster reagents

Best ADPGK / ADP-dependent glucokinase IHC Antibodies

A10813-1 has a human tonsil paraffin-section IHC image (catalog image caption); IF is listed without a figure, and human, mouse, and rat reactivity is listed (catalog applications/reactivity).

Real IHC data Immunohistochemical analysis of paraffin-embedded human tonsil. 1, Antibody was diluted at 1:200 (4° overnight). 2, Tris-EDTA, pH9.0 was used for antigen retrieval. 3, Secondary antibody was diluted at 1:200 (room temperature, 30min).

Anti-ADP-GK Antibody

Cat # A10813-1

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A10813-1 is listed for IHC and IF in human, mouse, and rat (catalog applications/reactivity). Its IHC image shows paraffin-embedded human tonsil at 1:200 primary dilution with Tris-EDTA pH 9.0 retrieval; no IF image is supplied (A10813-1 image captions).

Which to pick: For tissue IHC, choose A10813-1: its own caption documents paraffin-embedded human tonsil, but does not report the fixative (A10813-1 IHC image caption). For IF, A10813-1 lists a 1:200–1:1000 range; ICC is unlisted and no IF figure is supplied (catalog applications/dilutions/image captions). For cross-species work, A10813-1 lists human, mouse, and rat reactivity; it has a rabbit host and no specified clone, while its tissue image documents human tonsil only (catalog reactivity/host/clone; A10813-1 IHC image caption).

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

References

UniProt Consortium. UniProt entry Q9BRR6 (ADPGK_HUMAN, ADP-dependent glucokinase).
Human Protein Atlas. ADPGK tissue IHC expression (reliability: Approved).
Human Protein Atlas. ADPGK subcellular location (ICC-IF): Localized to the centrosome..
Human Protein Atlas. ADPGK antibody validation summary (2 antibodies).
Evaluation of tumorous LCP1 and ADPGK as predictive biomarker for immune-related adverse events in bone and soft tissue sarcomas treated with anti-PD-1 and anti-PD-L1 antibodies. BMC cancer 2025 — PMC11974013.
Prediction of immune-related adverse events in urological cancer during checkpoint inhibitor immunotherapy through immunohistochemical analysis of tumorous LCP1/ADPGK. Translational oncology 2026 — PMC13266032.
Multi-omics prediction of immune-related adverse events during checkpoint immunotherapy. Nature communications 2020 — PMC7532211.
ADP-dependent glucokinase controls metabolic fitness in prostate cancer progression. Military Medical Research 2023 — PMC10714548.
PubMed PMID:11230166 — UniProt-cited evidence.
PubMed PMID:14702039 — UniProt-cited evidence.
PubMed PMID:16303743 — UniProt-cited evidence.