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HK1 / Hexokinase-1 · IHC design guide
Design Immunohistochemistry for HK1
Plan HK1 paraffin-section IHC using neuronal cells as a high-staining reference and adipocytes as an undetected comparator (HPA tissue IHC). This guide covers fixation consistency, chromogenic detection and cell-level scoring (standard IHC practice).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
HK1 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 | Granular cytoplasm in tissue (HPA tissue IHC) | |
| Staining pattern | Widespread granular cytoplasmic staining across cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M01504-2) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Adipose tissue+1 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). Selected-image fixative and duration unreported (datasheet M01504-2); verify before use. | |
| Caveat | N-acetyl-D-glucosamine can shift HK1 to the cytosol (UniProt) | |
| Regulation | High in neurons; undetected in adipocytes (HPA tissue IHC) | |
| Isoform / epitope | 4 isoforms; 2 is erythrocyte-specific, 3/4 testis-specific; epitope unspecified (UniProt) |
Section 1
Recommended HK1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with published HK1 staining conditions from mouse tissue and mink uterus (PMC7658079; PMC4284318).
| Sample | Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet M01504-2) |
| Fixation | Image fixative and duration unreported (datasheet M01504-2); 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 M01504-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M01504-2) |
| Primary antibody | Mouse monoclonal (clone 4B7) anti-HK1, 0.5-1μg/ml (datasheet M01504-2) |
| Primary incubation | Overnight at 4 °C (datasheet M01504-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet M01504-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | HK1-positive staining in endocrine cells of appendix (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression with a granular pattern. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet: M01504-2). For mink uterus, the published alternative is sodium citrate at 90 °C for 20 min (PMC4284318).
Section 2
What Is the Expected HK1 Staining Pattern?
In paraffin-section IHC, expect HK1 mainly as granular cytoplasmic staining, with high signal in selected endocrine, epithelial, and neuronal cells (HPA: tissue IHC, Supported; medium consistency with RNA). Its mitochondrial outer-membrane association explains a punctate cytoplasmic pattern; cytosolic HK1 can also occur (UniProt P19367: localization). HK1 has no transmembrane segment (UniProt P19367: topology).
| Granular cytoplasmic staining in cerebral-cortex neurons or bronchial respiratory epithelium. | This fits the reported HK1 tissue pattern and high staining in these cell types (HPA: tissue IHC). Granules are compatible with mitochondrial association (UniProt P19367: localization). Judge the pattern within identified cells, since HPA describes staining across many tissues rather than a uniformly high signal in every cell (HPA: ubiquitous cytoplasmic profile; low tissue specificity). |
| Predominantly nuclear staining, with little cytoplasmic signal. | A nuclear-only result conflicts with the reported granular cytoplasmic IHC pattern and mitochondrial localization (HPA: tissue IHC; HPA: subcellular ICC-IF). Treat it as a possible staining artefact and review the matched control section, counterstain, and detection conditions (general IHC practice). Nuclear color alone does not establish HK1 localization. |
| Strong staining in adipocytes or chondrocytes while expected positive cells stain weakly. | HPA reports HK1 as not detected in adipocytes and chondrocytes in its sampled tissues (HPA: tissue IHC). Unexpected color in those cells warrants checks for antibody cross-reactivity or endogenous detection activity (general IHC practice). These cell-level observations must not be read as a claim that every cell in adipose or soft tissue is negative. |
| Broad, hazy color across cells and surrounding tissue, obscuring cytoplasmic granules. | This is hard to reconcile with HPA's granular cytoplasmic pattern (HPA: tissue IHC). Excess detection background or nonspecific antibody binding are possibilities (general IHC practice). Compare a no-primary control and examine whether color follows cell boundaries; a diffuse deposit is insufficient evidence for HK1 even in a tissue with reported positive cells. |
| No signal in a section containing recognizable cerebral-cortex neurons or bronchial respiratory epithelium. | Those cell types show high HK1 staining in HPA tissue IHC (HPA: cerebral cortex neuronal cells; bronchus respiratory epithelial cells). First confirm that the expected cells are present and assess the run controls (general IHC practice). A negative section cannot distinguish a technical failure from a sample-specific biological difference without a working positive control. |
💡Expected HK1 appearanceA convincing positive is granular cytoplasmic color in identifiable high-staining cells, such as cerebral-cortex neurons; isolated nuclear color or cell-free haze is suspect (HPA: tissue IHC; UniProt P19367: localization).
| Compartment and distribution | HK1 associates with the mitochondrial outer membrane and can relocate to the cytosol (UniProt P19367: localization). HPA describes granular cytoplasmic tissue staining (HPA: tissue IHC); a strictly nuclear pattern therefore needs investigation, while some less punctate cytoplasmic signal need not be excluded solely by shape. |
| Cell-specific comparison | HPA reports high staining in appendix endocrine cells, esophageal squamous epithelium, and hippocampal neurons, but low staining in liver cholangiocytes and pancreatic exocrine cells (HPA: tissue IHC). Score the named cell population rather than treating the whole tissue as one intensity category. |
| Isoforms and antibody epitope | UniProt lists four HK1 isoforms, including an erythrocyte-specific isoform 2 and testis-specific isoforms 3 and 4 (UniProt P19367: isoforms; tissue specificity). The supplied evidence gives no epitope or isoform coverage for the chosen antibody, so staining cannot be assigned to a particular isoform from location alone. |
| Strength of pattern evidence | HPA rates the tissue pattern Supported, with medium consistency between staining and RNA data; its three listed antibodies have IHC Supported status (HPA: tissue IHC; antibody validation). Use the pattern as a reference, while assessing the actual reagent and controls used in this experiment (general IHC practice). |
| IF/ICC: what localization is expected? | HPA reports enhanced mitochondrial localization by ICC-IF (HPA: subcellular ICC-IF). That observation supports the cytoplasmic-granule interpretation in IHC, but it does not specify an IF/ICC protocol or prove that every chromogenic granule is a mitochondrion (HPA: tissue IHC; subcellular ICC-IF). |
| Situation | Likely cause | Next action |
|---|---|---|
| No color in a known-positive cell population. | The result conflicts with high staining reported for cerebral-cortex neurons or bronchial respiratory epithelium (HPA: tissue IHC); a run failure remains possible (general IHC practice). | Confirm the cell population on the counterstained section, inspect the positive control, and verify the IHC-validated antibody and detection steps used for this run (general IHC practice). |
| Granules are faint but the positive control works. | Intensity varies by cell type: HPA reports low staining in cholangiocytes and pancreatic exocrine cells versus high staining in selected neuronal and epithelial cells (HPA: tissue IHC). | Compare like cell types and score localization alongside intensity; avoid calling a low-staining population negative solely because it is weaker than a high-staining control (HPA: tissue IHC). |
| Color is mainly nuclear. | Nuclear-dominant color conflicts with HPA's granular cytoplasmic IHC pattern and mitochondrial ICC-IF localization (HPA: tissue IHC; subcellular ICC-IF). | Check the no-primary control and whether the counterstain or chromogen is being misread; repeat interpretation using cytoplasmic signal in a known-positive cell population (general IHC practice; HPA: tissue IHC). |
| Adipocytes or chondrocytes stain strongly. | HPA reports these sampled cell types as not detected (HPA: tissue IHC). Cross-reactivity or endogenous detection activity may explain unexpected color (general IHC practice). | Review a no-primary control and detection-system controls, then compare the unexpected cells with a documented positive population on the same run (general IHC practice; HPA: tissue IHC). |
| Diffuse color masks cellular detail. | A widespread deposit does not match the reported granular cytoplasmic pattern (HPA: tissue IHC); nonspecific binding or detection background may contribute (general IHC practice). | Inspect a no-primary control, blocking and wash conditions, and detection exposure; score HK1 only where cellular cytoplasmic staining is distinguishable (general IHC practice; HPA: tissue IHC). |
| A tissue section contains both positive and apparently negative cells. | HPA reports cell-specific staining levels within tissues and an overall ubiquitous cytoplasmic profile, not identical intensity in all cells (HPA: tissue IHC). | Identify and score each cell population separately. Compare observed compartment and intensity with the matching HPA cell-level entry before labeling the entire section discordant (HPA: tissue IHC). |
Experiment evidence
IHC & IF images showing HK1 staining
M01504-2 · IHCIHC analysis of Hexokinase 1/HK1 using anti-Hexokinase 1/HK1 antibody (M01504-2). Hexokinase 1/HK1 was detected in paraffin-embedded section
M01504-2 · IHCIHC analysis of Hexokinase 1/HK1 using anti-Hexokinase 1/HK1 antibody (M01504-2). Hexokinase 1/HK1 was detected in paraffin-embedded section
M01504-2 · IFIF analysis of Hexokinase 1/HK1 using anti-Hexokinase 1/HK1 antibody (M01504-2). Hexokinase 1/HK1 was detected in immunocytochemical section
A01504-1 · IHCIHC analysis of Hexokinase 1/HK1 using anti-Hexokinase 1/HK1 antibody (A01504-1). Hexokinase 1/HK1 was detected in paraffin-embedded section
A01504-1 · IHCIHC analysis of Hexokinase 1/HK1 using anti-Hexokinase 1/HK1 antibody (A01504-1). Hexokinase 1/HK1 was detected in paraffin-embedded section
A01504-1 · IFIF analysis of Hexokinase 1/HK1 using anti-Hexokinase 1/HK1 antibody (A01504-1). Hexokinase 1/HK1 was detected in immunocytochemical section
M01504-1 · IHCIHC analysis of Hexokinase 1/HK1 using anti-Hexokinase 1/HK1 antibody (M01504-1). Hexokinase 1/HK1 was detected in paraffin-embedded section
M01504-1 · IHCIHC analysis of Hexokinase 1/HK1 using anti-Hexokinase 1/HK1 antibody (M01504-1). Hexokinase 1/HK1 was detected in paraffin-embedded section
M01504-1 · IFIF analysis of Hexokinase 1/HK1 using anti-Hexokinase 1/HK1 antibody (M01504-1). Hexokinase 1/HK1 was detected in immunocytochemical section
A01504-2 · IHCImmunohistochemistry of Hexokinase 1 in rat stomach tissue with Hexokinase 1 antibody at 5 μg/mL.
A01504-2 · IFImmunofluorescence of Hexokinase 1 in rat stomach tissue with Hexokinase 1 antibody at 20 μg/mL.
M01504 · IHCImmunohistochemical analysis of paraffin-embedded human lung carcinoma, using Hexokinase 1 Antibody.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000156515-HK1/tissue/cerebral+cortex#imid_3315864
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000156515-HK1/tissue/colon#imid_3315806
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000156515-HK1/tissue/liver#imid_3315799
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000156515-HK1/tissue/kidney#imid_3315792
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB010052; green = HK1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB010052; green = HK1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for HK1 IHC & IF
🧪Run bronchus first: respiratory epithelial cells should stain strongly (HPA: High in bronchus respiratory epithelial cells). Use adipose tissue as the negative comparator (HPA: adipocytes Not detected); on the bronchus slide, cells without specific HK1 staining should show only background chromogen, but the supplied HPA row does not establish a named internal negative cell type (HPA: bronchus respiratory epithelial cells High).
Positive control tissue: Appendix (Endocrine cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show HK1 in A-431, U-251MG, U2OS, hTERT-RPE1 (serum starved), with annotated localisation: Mitochondria (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a matched mouse IgG isotype control, and HK1 knockout tissue as a biological negative (caption: mouse primary antibody; standard IHC practice). Quench endogenous peroxidase and assess endogenous biotin background in bronchus sections before interpreting DAB staining (caption: biotinylated secondary, SABC and DAB; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected M01504-2 paraffin-section caption does not state a fixative (caption: fixative unreported). The caption supports testing heat retrieval in EDTA at pH 8.0 as a starting condition, with retrieval verified in bronchus (caption: lung cancer paraffin-section IHC). Frozen-section performance and any advantage for IF are unreported; bronchial tissue autofluorescence may complicate IF interpretation (HPA: ICC-IF mitochondrial localization; standard IF practice).
HPA tissue IHC evidence for HK1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — 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 |
|---|---|---|---|---|
| Appendix | Endocrine cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Colon | Endocrine cells | High | Protein (IHC) | HPA → |
Section 3
Advanced HK1 IHC Tips
Troubleshoot HK1 staining in paraffin sections by checking retrieval, cell type, cytoplasmic pattern and controls before comparing signal intensity (datasheet M01504-2; HPA tissue IHC).
Which retrieval conditions should I try first for weak HK1 staining?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet M01504-2). The catalog antibody produced chromogenic staining in a paraffin-embedded human lung cancer section after this retrieval, with 1 μg/mL primary antibody incubated overnight at 4°C (datasheet M01504-2). If signal remains weak, compare retrieval duration on adjacent sections while holding antibody concentration and detection conditions constant (standard IHC practice). Include a section with an expected positive cell population, such as bronchial respiratory epithelial cells, to distinguish retrieval failure from a genuinely low-signal region (HPA: High in bronchial respiratory epithelial cells).
Could fixation explain weak or uneven HK1 staining?
The selected paraffin-section caption does not state a fixative, so HK1-specific sensitivity to fixation is unknown for this antibody (datasheet M01504-2). Record the actual fixative, fixation duration and processing history for each block before comparing staining across specimens (standard IHC practice). If weak staining tracks with processing history, compare adjacent sections using the documented EDTA pH 8.0 retrieval and identical detection conditions (datasheet M01504-2; standard IHC practice). Keep tissue thickness, primary incubation and development consistent, then assess whether the change improves cellular pattern as well as intensity (standard IHC practice).
What cellular pattern should convincing HK1 chromogenic staining show?
Look for cytoplasmic staining with a granular pattern in appropriately preserved cells (HPA tissue IHC: ubiquitous granular cytoplasmic expression). HK1 is associated with the mitochondrial outer membrane and can also occur in the cytosol; its mitochondrial-binding peptide promotes membrane association (UniProt P19367 localisation). A granular cytoplasmic pattern is therefore plausible, but a DAB image alone cannot establish that each granule is mitochondrial (HPA tissue IHC; standard IHC interpretation). Compare cell types within the same section and use an orthogonal localisation assay if a mitochondrial-versus-cytosolic conclusion matters (standard IHC practice).
Can this stain distinguish HK1 isoforms or reveal its mitochondrial-binding region?
HK1 has 4 annotated isoforms, including an erythrocyte-specific isoform 2 and testis-specific isoforms 3 and 4 (UniProt P19367 isoforms). The supplied catalog caption identifies HK1 staining but does not specify the antibody epitope or establish isoform discrimination (datasheet M01504-2). Interpret chromogenic signal as antibody-detected HK1 unless epitope mapping or isoform-specific validation supports a narrower claim (standard IHC interpretation). The mitochondrial-binding peptide promotes outer-membrane association, while the protein has no transmembrane segment; staining pattern alone cannot identify which region the antibody recognizes (UniProt P19367 topology and localisation; standard IHC interpretation).
How should I assess HK1 by IF alongside cell-type and mitochondrial markers?
Use this as a separate IF validation experiment: the supplied antibody evidence describes chromogenic staining in a paraffin section, not an IF protocol (datasheet M01504-2). Multiplex HK1 with a validated marker for the expected cell population and, when localisation matters, a mitochondrial marker (HPA: High in bronchial respiratory epithelial cells; UniProt P19367 localisation; standard IF practice). Choose fluorophores after checking unstained tissue autofluorescence, and include single-label controls for channel bleed-through (standard IF practice). Test gentle permeabilisation for access to the cytosol-associated, outer-membrane HK1 pool; the antibody epitope and its accessibility are unspecified, so verify the resulting pattern experimentally (UniProt P19367 topology and localisation; standard IF practice).
How can I separate widespread HK1 staining from chromogenic background?
HK1 can show widespread granular cytoplasmic staining, so signal in many cell types is not by itself evidence of background (HPA tissue IHC: ubiquitous granular cytoplasmic expression). Compare a no-primary control and an expected low-signal population, such as adipocytes, with the stained section (HPA: Not detected in adipocytes; standard IHC practice). Include a peroxidase-blocking step and check the no-primary control before attributing diffuse brown deposits to HK1 (standard chromogenic IHC practice). The selected caption used biotinylated secondary antibody, streptavidin-biotin detection and DAB, making detection-system controls relevant when staining is unexpectedly widespread (datasheet M01504-2; standard IHC practice).
How should I score HK1 staining across heterogeneous sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define cell populations and analysis regions before scoring, because HK1 staining varies by cell type even within its broadly expressed tissue profile (HPA tissue IHC). For cellular staining, report the percentage of positive cells and an intensity-based H-score using the same scoring thresholds across sections (standard IHC practice). If cell abundance is the endpoint, report positive-cell density per mm² of viable tissue and state how regions were selected (standard IHC practice). Normalise positive counts to the relevant total cell population, and keep retrieval, antibody incubation and DAB development consistent across compared specimens (standard IHC practice).
Which findings would make an apparent HK1-positive region questionable?
Convincing staining should align with a granular cytoplasmic pattern in intact cells, with cell-type differences assessed against the expected tissue profile (HPA tissue IHC). Predominantly nuclear signal is discordant with the supplied mitochondrial outer-membrane and cytosolic localisation evidence (UniProt P19367 localisation). Treat staining confined to section edges or necrotic areas cautiously, and compare it with matched controls and well-preserved regions (standard IHC practice). Check the no-primary and peroxidase-block controls before calling diffuse DAB deposits positive, since endogenous enzyme activity can contribute chromogenic signal (standard IHC practice).
Boster reagents
Best HK1 / Hexokinase-1 IHC Antibodies
Five anti-HK1 antibodies have IHC images from human lung cancer or carcinoma, mouse brain, and rat stomach; catalog reactivity includes human, mouse, and rat, and IF images are also available (catalog image captions; catalog reactivity).
Real IHC data
Anti-Hexokinase 1/HK1 Antibody ® (monoclonal, 4B7)
Cat # M01504-2
Real IHC data
Anti-Hexokinase 1/HK1 Antibody ®
Cat # A01504-1
Real IHC data
Anti-Hexokinase 1/HK1 Antibody ® (monoclonal, 2I4)
Cat # M01504-1
Real IHC data
Anti-Hexokinase 1 HK1 Antibody
Cat # A01504-2
Real IHC data
Anti-Hexokinase 1 HK1 Rabbit Monoclonal Antibody
Cat # M01504
M01504-2 and M01504-1 show paraffin-section IHC in human lung cancer, while A01504-1 shows paraffin-section IHC in mouse brain (respective IHC captions). A01504-2 shows IHC in rat stomach, and M01504 shows paraffin-section IHC in human lung carcinoma (respective IHC captions).
Which to pick: For tissue IHC, choose M01504-2 for human lung cancer or A01504-1 for mouse brain: their paraffin-section captions each specify EDTA pH 8.0 retrieval and 1 μg/mL primary antibody overnight at 4°C (respective IHC captions). For IF/ICC, M01504-2 lists both applications and has IF images from cells and a paraffin section of human lung cancer (catalog applications; M01504-2 IF captions). For cross-species IHC, A01504-1 lists human, mouse, and rat reactivity and has IHC images from all three species; its paraffin-section captions do not report the fixative (catalog reactivity; A01504-1 IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P19367 (HXK1_HUMAN, Hexokinase-1).
- Human Protein Atlas. HK1 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. HK1 subcellular location (ICC-IF): Localized to the mitochondria..
- Human Protein Atlas. HK1 antibody validation summary (3 antibodies).
- Direct neuronal glucose uptake heralds activity-dependent increases in cerebral metabolism. Nature communications 2015 — PMC4410436.
- Role of human tissue kallikrein in gastrointestinal stromal tumour invasion. British journal of cancer 2010 — PMC2990601.
- TAB1 regulates glycolysis and activation of macrophages in diabetic nephropathy. Inflammation research : official journal of the European Histamine Research Society ... [et al.] 2020 — PMC7658079.
- Uterine glycogen metabolism in mink during estrus, embryonic diapause and pregnancy. The Journal of reproduction and development 2014 — PMC4284318.
- PubMed PMID:3207429 — UniProt-cited evidence.
- PubMed PMID:9531504 — UniProt-cited evidence.
- PubMed PMID:15164054 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for HK1 (Hexokinase-1). Expected localisation Granular cytoplasm in tissue (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M01504-2). Validated IHC antibody: M01504-2 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Endocrine cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet M01504-2); 2. Fixation: Image fixative and duration unreported (datasheet M01504-2); 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 M01504-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M01504-2); 8. Primary antibody: Mouse monoclonal (clone 4B7) anti-HK1, 0.5-1μg/ml (datasheet M01504-2); 9. Primary incubation: Overnight at 4 °C (datasheet M01504-2). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific). Selected-image fixative and duration unreported (datasheet M01504-2); verify before use.; Caveat: N-acetyl-D-glucosamine can shift HK1 to the cytosol (UniProt); Isoform / epitope: 4 isoforms; 2 is erythrocyte-specific, 3/4 testis-specific; epitope unspecified (UniProt); Regulation: High in neurons; undetected in adipocytes (HPA tissue IHC). Evidence: UniProt P19367, HPA tissue IHC · Supported, HPA ICC-IF · mitochondria., Datasheet M01504-2, PMC7658079, PMC4284318. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/HK1/immunohistochemistry-ihc-guide.