C1QBP / Complement component 1 Q subcomponent-binding protein, mitochondrial · IHC design guide

Design Immunohistochemistry for C1QBP

Plan C1QBP chromogenic IHC in paraffin sections using the granular cytoplasmic tissue pattern as a reference (HPA tissue IHC). A catalog antibody example uses a 1:50 primary dilution (datasheet M01439); germinal center cells stain strongly, while cholangiocytes are not detected (HPA tissue IHC).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
Immunohistochemistry protocol sheet for C1QBP (IHC for C1QBP): expected localisation Granular cytoplasmic tissue staining (HPA tissue IHC), antibody M01439, validated IHC image, and IHC protocol steps
Printable C1QBP IHC protocol sheet — expected localisation Granular cytoplasmic tissue staining (HPA tissue IHC), antibody M01439, controls and protocol steps. Open the full C1QBP IHC guide →

C1QBP 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 cytoplasmic tissue staining (HPA tissue IHC)
Staining pattern Granular cytoplasm; strong in germinal center cells (HPA tissue IHC)
Antigen retrieval EDTA pH 8.0 HIER, heat-mediated (datasheet M01439)
Positive control ⓘ Appendix+4 more · see all
Negative control ⓘ Liver+1 more · see all
Important caveats
Reasons your staining may differ from the expected pattern.
Fixation Keep formalin fixation consistent across paraffin sections (standard IHC practice; not target-specific)
Caveat A secreted variant may complicate tissue staining patterns (HPA tissue IHC)
Regulation Activated lymphocytes secrete C1QBP (UniProt)
Isoform / epitope No annotated isoforms; mature chain spans 74–282 and has no transmembrane segment, so there is no defined extracellular versus cytoplasmic epitope split (UniProt)
Section 1

Recommended C1QBP IHC & IF Protocols

The catalog antibody has a separate IHC-P protocol (datasheet M01439). The published options below cover rat brain free-floating sections (PMC6418184) and mouse paraffin-embedded lung sections (PMC13069790).

Recommended immunohistochemistry (IHC-P) protocol parameters
SampleParaffin-embedded human breast cancer tissue; fixative not specified (datasheet M01439)
FixationImage fixative and duration unreported (datasheet M01439); verify before use.
Sectioning4–5 µm sections on charged slides (standard)
DeparaffinisationXylene, graded ethanol series to water (standard)
Antigen retrievalHeat retrieval: EDTA pH 8.0 (datasheet M01439); 20 min, 95–100 °C (standard)
Peroxidase block3% H2O2, 10 min, room temperature (standard)
Blocking10% goat serum (datasheet M01439)
Primary antibodyRabbit monoclonal (clone 17C69) anti-C1QBP, 1:50-1:200 (datasheet M01439)
Primary incubationOvernight at 4 °C (datasheet M01439)
DetectionHRP-conjugated secondary, DAB chromogen (datasheet M01439)
CounterstainHematoxylin, blue, dehydrate and mount (standard)
Expected resultC1QBP-positive staining in germinal center 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 noteFor paraffin sections, start with heat-mediated EDTA pH 8.0 retrieval (datasheet M01439); the published mouse lung protocol reports retrieval without specifying its buffer (PMC13069790).
Section 2

What Is the Expected C1QBP Staining Pattern?

In paraffin-section IHC, expect widespread granular cytoplasmic C1QBP staining, including strong staining in selected germinal center and glandular cells (HPA: supported tissue IHC; high in appendix germinal center and breast glandular cells). C1QBP is predominantly mitochondrial but also has reported surface, nuclear, cytoplasmic and secreted locations; it has no transmembrane segment (UniProt Q07021 localization and topology).

What am I looking at on my slide?
Granular cytoplasmic signal is strong in appendix or lymph-node germinal center cells; nearby cells vary in intensity.This fits the reported tissue pattern: C1QBP expression is widespread, with high staining in these germinal center cells (HPA: tissue IHC). Score the named cells rather than treating every cell in the section as an equally strong positive control.
Strong, uniform nuclear-only staining replaces the expected granular cytoplasmic pattern across the section.Treat this as a possible staining artefact and check controls and morphology (HPA: granular cytoplasmic tissue pattern; standard IHC practice). Nuclear and nucleolar localization is reported, so an isolated nuclear signal cannot be declared false solely from its compartment (UniProt Q07021 localization).
The strongest chromogen appears in HPA-negative cholangiocytes or skeletal myocytes, while expected positive cells are weak.Question antibody specificity or endogenous detection activity, especially if staining ignores cell boundaries (HPA: C1QBP not detected in those named cell types; standard IHC practice). These observations concern specific cells, not the entire liver or skeletal-muscle section.
Color spreads diffusely through stroma, extracellular space or most cells without a discernible granular pattern.Interpret this as background until a no-primary control and detection controls are clean (standard IHC practice). Secreted C1QBP is reported, but secretion alone does not validate diffuse chromogen in a paraffin section (UniProt Q07021 localization).
No staining is visible in appendix germinal center cells or breast glandular cells.A failed positive control is more likely than a meaningful negative result in the test section; review staining and tissue preservation before scoring it (HPA: high staining in those cells; standard IHC practice). HPA support does not guarantee signal in every specimen.
💡Expected C1QBP appearanceCall a result positive when identifiable cells show granular cytoplasmic chromogen, preferably strong in HPA-high germinal center or breast glandular cells; widespread featureless color or dominant staining of HPA-undetected cell types calls for control checks (HPA: tissue IHC; standard IHC practice).
How each factor affects the staining
Tissue and cell selectionAppendix and lymph-node germinal center cells, breast glandular cells, intestinal endocrine cells and ovarian follicle cells are reported high; cholangiocytes and skeletal myocytes are not detected (HPA: tissue IHC). Compare the annotated cell population, not whole-organ color.
Compartment and assayParaffin IHC is described as granular cytoplasmic (HPA: tissue IHC), while ICC-IF shows mainly plasma membrane and additional mitochondrial localization (HPA: supported subcellular ICC-IF). UniProt describes predominantly mitochondrial localization plus other compartments (UniProt Q07021 localization). Interpret each assay in its own context.
Processing and secreted proteinThe annotated mature chain spans residues 74–282, and a secreted form is reported (UniProt Q07021 processing and localization). The payload gives no antibody epitope or tissue-specific shedding result, so neither processing nor secretion predicts a particular chromogen pattern.
Antibody evidenceThe listed antibody HPA026483 has Supported IHC and ICC status (HPA: antibody validation). The tissue profile is also Supported through consistency with RNA data, with a secretion caveat (HPA: tissue-IHC reliability). These labels support interpretation but do not establish performance for every staining condition.
IF/ICC Q: Should membrane signal be accepted?Yes, when the IF/ICC image and controls support it: HPA reports a supported plasma-membrane main location and additional mitochondrial location (HPA: subcellular ICC-IF). Use the separate IF/ICC guide for that assay; this paraffin-IHC section makes no IF protocol recommendation.
Fixation and retrieval evidenceTarget-specific fixation sensitivity and a retrieval condition are unreported in the supplied UniProt and HPA records. Retrieval optimization is a general paraffin-IHC workflow decision (standard IHC practice); no C1QBP-specific gain or loss can be predicted from these sources.
Why is my staining missing, weak or wrong?
SituationLikely causeNext action
Known HPA-high cells are unstained.The staining run, antigen retrieval or detection may have failed; the observation alone does not identify which step (HPA: high in the named positive cells; standard IHC practice).Confirm tissue morphology and a working positive control, then review retrieval, antibody incubation and detection in sequence (standard IHC practice). Do not score other sections negative until the control works.
Chromogen is widespread and featureless.Nonspecific binding or detection background may obscure the reported granular cytoplasmic pattern (HPA: tissue IHC; standard IHC practice).Compare a no-primary control; review blocking, washes, primary-antibody concentration and detection exposure (standard IHC practice). Retain a result only if cell boundaries and the expected granular pattern remain interpretable.
Color appears in the no-primary control.Endogenous detection activity or reagent background can produce signal without the primary antibody (standard chromogenic IHC practice).Address the detection system's endogenous activity and repeat the control alongside the test slide (standard chromogenic IHC practice). Do not assign control-positive color to C1QBP.
Cholangiocytes or skeletal myocytes stain strongly.Those named populations are not detected in the HPA tissue profile; cross-reactivity or background is plausible, although specimen variation cannot be resolved from this observation alone (HPA: tissue IHC; standard IHC practice).Check cell identity, no-primary signal and staining in an HPA-high cell population before interpreting the unexpected signal (HPA: tissue IHC; standard IHC practice).
Nuclear-only staining dominates a paraffin section.It differs from the HPA granular cytoplasmic tissue pattern, though nuclear and nucleolar C1QBP are reported in other contexts (HPA: tissue IHC; UniProt Q07021 localization).Review morphology and controls, then compare a known positive tissue stained in the same run (standard IHC practice). Document a reproducible nuclear signal separately from the expected tissue-IHC pattern.
Membrane signal in IF/ICC seems inconsistent with cytoplasmic IHC.The reported assay patterns differ: HPA tissue IHC is granular cytoplasmic, while HPA ICC-IF mainly localizes C1QBP to the plasma membrane (HPA: tissue IHC and supported subcellular ICC-IF).Judge each image against its assay-specific HPA pattern and its controls; do not transfer the IF/ICC membrane expectation into a paraffin-IHC scoring rule (HPA: tissue IHC and subcellular ICC-IF; standard interpretation practice).

Sample controls for C1QBP IHC & IF

🧪Run breast first: glandular cells should stain (HPA: breast glandular cells High). Run liver as a negative tissue, focusing on cholangiocytes (HPA: liver cholangiocytes Not detected); within the breast slide, use cells lacking signal as a background comparator if present, since no specific negative cell population is identified (HPA: breast glandular cells High).
Positive control tissue: Appendix (Germinal center cells, HPA High)
Negative control tissue: Liver (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show C1QBP in A-431, U-251MG, U2OS, KOLF2.1J, with annotated localisation: Plasma membrane (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host-matched rabbit IgG isotype controls (selected-SKU caption: rabbit anti-C1QBP), plus C1QBP knockout tissue or a validated peptide-block control where available (standard IHC practice). Block endogenous peroxidase and check for background staining in the breast section before interpreting DAB signal (selected-SKU caption: breast tissue, HRP/DAB detection; standard IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected-SKU paraffin-section caption does not state the fixative (selected-SKU caption: fixative not stated). The example uses heat-mediated EDTA retrieval at pH 8.0, but does not establish that retrieval is required (selected-SKU caption: EDTA pH 8.0); the supplied evidence does not establish whether frozen sections or IF are easier. Assess breast-section background alongside glandular-cell staining, particularly with HRP/DAB detection (selected-SKU caption: breast tissue, HRP/DAB detection; standard IHC practice).

HPA tissue IHC evidence for C1QBP

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

Positive expression · recommended positive controls

TissueCell typeLevelEvidenceSource
Appendix Germinal center cells High Protein (IHC) HPA →
Breast Glandular cells High Protein (IHC) HPA →
Colon Endocrine cells High Protein (IHC) HPA →
Lymph node Germinal center cells High Protein (IHC) HPA →
Ovary Follicle cells High Protein (IHC) HPA →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
Liver Cholangiocytes Not detected Protein (IHC) HPA →
Skeletal muscle Myocytes Not detected Protein (IHC) HPA →
Section 3

Advanced C1QBP IHC Tips

Use the catalog antibody’s paraffin-section workflow as the starting point, then assess staining against C1QBP’s documented distribution and compartment patterns.

Which retrieval conditions should I start with for C1QBP in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet M01439). The selected paraffin-section image used that retrieval before 10% goat serum blocking and a 1:50 primary-antibody incubation overnight at 4°C (datasheet M01439). If staining is weak, first check heating consistency and whether sections remained covered by buffer, then compare a shorter or longer retrieval within the same EDTA workflow (standard IHC practice). Score any change against both expected granular cytoplasmic staining and unwanted diffuse background, using the same tissue and detection conditions for the comparison (HPA tissue IHC; standard IHC practice).
How should I troubleshoot fixation-related loss of C1QBP staining?
The selected tissue image identifies a paraffin-embedded section but does not report its fixative, so target-specific fixation sensitivity is unknown (datasheet M01439). Record the fixative, fixation duration, section age, and retrieval run for each specimen before comparing staining (standard IHC practice). When signal varies between batches, process adjacent sections with the same EDTA pH 8.0 retrieval and 1:50 primary-antibody conditions used in the selected image (datasheet M01439). Compare staining in matched cell populations and include a known staining control in each run; neither a tissue-expression pattern nor a topology annotation establishes a C1QBP-specific fixation effect (standard IHC practice; HPA tissue IHC; UniProt Q07021 topology).
Should C1QBP staining be granular cytoplasmic, membranous, or nuclear?
In tissue IHC, the reported pattern is ubiquitous granular cytoplasmic staining, which is a useful starting expectation for paraffin sections (HPA tissue IHC). C1QBP is predominantly mitochondrial, while nuclear, nucleolar, cell-surface, secreted, and cytoplasmic locations are also reported (UniProt Q07021 subcellular). Review cell boundaries and counterstained nuclei at higher magnification before assigning a membranous or nuclear pattern, particularly where dense granular staining overlaps those structures (standard IHC practice). Nucleolar localisation has a specific reported association with POLGARF coexpression, so an isolated nuclear signal should prompt orthogonal localisation evidence rather than an automatic biological assignment (UniProt Q07021 subcellular; PubMed:32958672; standard IHC practice).
Could processing or epitope position explain discordant C1QBP staining?
C1QBP is recorded as a 282-residue precursor with a processed chain spanning residues 74–282, and no isoforms are annotated (UniProt Q07021 processing; UniProt Q07021 isoforms). Its listed modifications include phosphoserine at 87, phosphotyrosine at 188, and phosphoserine at 201 and 205 (UniProt Q07021 modified residues). The selected image does not identify the antibody epitope, so do not attribute a staining difference to cleavage or a modification without epitope information and direct testing (datasheet M01439; standard IHC practice). If discordance persists, compare the same regions with an independently validated antibody targeting a documented different epitope, while keeping retrieval and scoring consistent (standard IHC practice).
How can IF help check an ambiguous C1QBP IHC pattern?
Use IF as a separate localisation check and multiplex C1QBP with a marker for the cell population being assessed; keep the chromogenic paraffin result as the IHC reference (standard IF practice; datasheet M01439). Choose spectrally separated fluorophores and inspect unstained tissue to place the weaker signal in a channel with less autofluorescence (standard IF practice). For an intracellular mitochondrial or nuclear epitope, optimise permeabilisation after fixation; for a proposed surface-facing epitope, assess staining without permeabilisation and confirm accessibility with an appropriate control (UniProt Q07021 subcellular; standard IF practice). Because the antibody epitope and IF fixation conditions are not supplied, neither the paraffin retrieval nor the IHC incubation establishes an IF protocol for this antibody (datasheet M01439).
What should I check when C1QBP DAB staining looks diffuse?
The selected paraffin-section workflow used 10% goat serum blocking, a 1:50 primary-antibody dilution, peroxidase-based detection, and DAB (datasheet M01439). Check a no-primary section for secondary-reagent or detection-system staining, and include a peroxidase-block step when using HRP/DAB (standard IHC practice). Compare shorter primary incubation or a more dilute primary antibody with the documented overnight incubation at 4°C, keeping retrieval and DAB development matched (datasheet M01439; standard IHC practice). Genuine tissue staining is reported as granular cytoplasmic, so diffuse haze across cells, stroma, and section edges needs investigation before it is scored as C1QBP (HPA tissue IHC; standard IHC practice).
How should I score heterogeneous C1QBP staining across a section? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and anatomical region before scoring, because C1QBP tissue IHC shows a granular cytoplasmic pattern across many tissues (HPA tissue IHC; standard IHC practice). Report an H-score as the sum of each intensity category multiplied by its percentage of cells, or report percentage-positive cells using a prespecified threshold (standard IHC practice). For spatial counts, report positive-cell density per mm² of viable tissue and specify whether stromal and epithelial cells were analysed separately (standard IHC practice). Normalise comparisons to the same eligible cell population or viable tissue area, and hold retrieval, antibody incubation, detection, and image-acquisition settings constant across samples (standard IHC practice; datasheet M01439).
How can I distinguish true C1QBP positivity from artefact?
First look for cellular granular cytoplasmic staining, the reported tissue-IHC pattern, and assess whether the stained cells match the population being scored (HPA tissue IHC; standard IHC practice). A breast glandular-cell sample can provide a reported high-expression reference, while liver cholangiocytes are reported as not detected; use these observations as contextual controls, not absolute cutoffs (HPA tissue IHC). Investigate staining confined to cut edges, folds, necrotic regions, or no-primary sections before calling it positive (standard IHC practice). Membrane or nucleolar staining can be biologically plausible, but needs stronger compartment evidence because mitochondrial localisation predominates and nucleolar localisation is linked to POLGARF coexpression (UniProt Q07021 subcellular; PubMed:32958672).
Boster reagents

Best C1QBP / Complement component 1 Q subcomponent-binding protein, mitochondrial IHC Antibodies

Three anti-C1QBP antibodies have paraffin-section IHC images from human, mouse and rat tissues; PB9921 also has an IF/ICC cell image (catalog image captions).

Real IHC data IHC analysis of C1QBP using anti-C1QBP antibody (M01439). C1QBP was detected in a paraffin-embedded section of human breast cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-C1QBP Antibody (M01439) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Anti-GC1q R Rabbit Monoclonal Antibody
Cat # M01439
Real IHC data IHC analysis of C1QBP using anti-C1QBP antibody (PB9921). C1QBP was detected in paraffin-embedded section of human tonsil tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-C1QBP Antibody (PB9921) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Anti-GC1q R/C1QBP Antibody ®
Cat # PB9921
Real IHC data IHC analysis of C1QBP using anti-C1QBP antibody (M01439-1). C1QBP was detected in a paraffin-embedded section of human breast cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-C1QBP Antibody (M01439-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Anti-C1QBP Rabbit Monoclonal Antibody
Cat # M01439-1

M01439 has IHC images from human breast and pancreas cancer sections, mouse midbrain and rat midbrain (M01439 image captions); M01439-1 has images from human breast and pancreas cancer sections and rat cerebellum (M01439-1 image captions). PB9921 has IHC images from human tonsil, mouse spleen and rat spleen, plus an IF/ICC image from U20S cells (PB9921 image captions).

Which to pick: For paraffin-section IHC, choose M01439 or M01439-1 if you want a rabbit monoclonal, with each shown using EDTA retrieval at pH 8.0 and 1:50 primary dilution in its own IHC image; the fixative is unreported (M01439 and M01439-1 catalog entries and image captions). Choose PB9921 for IF/ICC because that application and a cell image are documented, with 2 μg/mL primary dilution in the IF caption (PB9921 catalog entry and IF image caption). For cross-species paraffin-section IHC, M01439 and PB9921 each have human, mouse and rat tissue images; M01439-1 lists reactivity with all three species but has no mouse IHC image in this payload (catalog reactivity lists and image captions).

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

References

  1. UniProt Consortium. UniProt entry Q07021 (C1QBP_HUMAN, Complement component 1 Q subcomponent-binding protein, mitochondrial).
  2. Human Protein Atlas. C1QBP tissue IHC expression (reliability: Supported).
  3. Human Protein Atlas. C1QBP subcellular location (ICC-IF): Mainly localized to the plasma membrane. In addition localized to the mitochondria..
  4. Human Protein Atlas. C1QBP antibody validation summary (1 antibodies).
  5. Complement C1q binding protein regulates T cells' mitochondrial fitness to affect their survival, proliferation, and anti-tumor immune function. Cancer science 2022 — PMC8898709.
  6. Complement component 1q subcomponent binding protein in the brain of the rat. Scientific reports 2019 — PMC6418184.
  7. C1QBP Associated With Immune Infiltration Predicts Poor Prognosis in Lung Adenocarcinoma. Cancer informatics 2026 — PMC12901909.
  8. Complement protein C1q induces endothelial apoptosis through C1qBP regulation of mitochondrial function in lung endothelial cells in systemic sclerosis. Molecular medicine (Cambridge, Mass.) 2026 — PMC13069790.
  9. PubMed PMID:8262387 — UniProt-cited evidence.
  10. PubMed PMID:8195709 — UniProt-cited evidence.
  11. PubMed PMID:11278463 — UniProt-cited evidence.