HemoVoid™ - Hemoglobin Depletion Reagent Kit
HemoVoid™ , a silica-based protein enrichment matrix, removes hemoglobin from erythrocyte lysate samples while concentrating low abundance, and/or low molecular weight proteins. The HemoVoid™ protocol uses mild buffers; the protocol conditions are so gentle that native enzyme activity is retained in elution fractions.
HemoVoid™ derives from a silica-based library of individual mixed-mode ligand combinations (ionic, hydrophobic, aromatic, polymer). The library was designed to facilitate weak binding of proteins, allowing for rapid elution from the matrix without any foreknowledge of the variety of proteins contained in the starting sample. HemoVoid™ depletes hemoglobin from red cell lysates while improving the resolution of less abundant blood proteins.
| Product | Size | Total Sample Processed | Item No. |
|---|---|---|---|
| HemoVoid™ | 10 preps | 10 x 300 µl | HVK-10 |
| HemoVoid™ | 50 preps | 50 x 300 µl | HVK-50 |
| HemoVoid™ | 100 Preps | 100 x 300 µl | HVK-100 |
| Items Required | 10 Prep | 50 Prep | 100 Prep | Reagent |
|---|---|---|---|---|
| HemoVoid™ | 0.5 gram | 2.5 grams | 5.0 grams | Supplied |
| Binding Buffer HVBB, PH 6.0 | 8 ml | 40 ml | 80 ml | Supplied |
| Wash Buffer HVWB, PH 7.0 | 15 ml | 75 ml | 150 ml | Supplied |
| Elution Buffer HVEB, PH 9.8 | 3 ml | 15 ml | 30 ml | Supplied |
| SpinX Centrifuge tube filters | 10 | 50 | 100 | Supplied |
Selectionof HemoVoid™ References:
Dried BloodCard/Spot Proteomics
HannahGing, Elin Davies, Claire E. Eyers, Louise Oni, Andrew Chetwynd. HAEMOGLOBININTERFERENCE IN DRIED BLOOD SPOT PROTEOMICS FOR CHRONIC KIDNEYDISEASE , Poster 2025 World ProteomeCongress, Toronto Canada. The study The HemoVoid™ workflow achievedthe highest coverage with the least variability compared toalternatives, and helped in the identification of potentialclinically relevant biomarkers in CKD patients receivinghaemodialysis, using dried blood cards.
Red Blood Cells (RBCs) /Parkinson’s Disease / α-Synuclein
ParraM.A., Snyder M.P. Parkinson’s Disease biomarker discovery viaLC-MS/MS characterization and quantification of alpha synuclein PTMsin erythrocytes, Poster 2025 ASMS Conference, Baltimore MD. The studyconcludes that while alpha synuclein proteoforms are analyticallychallenging to characterize by Mass Spectrometry, comparedto alternatives, the HemoVoid™ on-bead digest sample prep method,detected phospoS87 even in healthy patients.
Red BloodCells (RBCs) / Other Applications
Xi-LongWang, et al. (2025 ) ProteomeProfiling of Red Blood Cells from Patients with COPD Links ProteasomeActivation with Abnormal Cell Morphology and Function ,International Journal of Chronic Obstructive Pulmonary Disease, ,3279-3294, DOI: 10.2147/COPD.S531220 Using HemoVoid™ for Hemoglobin depletion, the study identified 160 differentiallyabundant proteins (DAPs) of which 70 were up-regulated, and 90down-regulated in the RBCs of COPD patients.
RedBlood Cells (RBCs) / Parkinson’s Disease / α-Synuclein
Cao,Chan, et al. " Deeplearning-assisted single-molecule detection of proteinpost-translational modifications with a biologicalnanopore ." bioRxiv (2023):2023-09. The authors demonstrate the ability of a biologicalnanopore, to detect and distinguish α-synuclein-derived peptidesbearing single or multiple PTMs, occurring at different positions andin various combinations. To deplete Hemoglobin, the article states“Briefly, RBCs… treated using the HemoVoid kit, … to remove hemoglobin but also to enrich low abundantproteins such as α-synuclein.”
Klatt,Stephan, et al. " Optimizingred blood cell protein extraction for biomarker quantitation withmass spectrometry ." Analyticaland Bioanalytical Chemistry (2020):1-14.
Thearticle describes the advantage of HemoVoid™ in detection of low abundance proteins when comparing their amounts(in percent) between four alternative extraction conditions, stating“… Most peptides, following HemoVoid™ extraction, showedion abundances ranging between 1.00E+5 and 1.00E+6 (31%). Incomparison to this, fewer peptides (10–23%) were within this rangefollowing extraction with all other protocols”. With respect topotential biomarkers for Parkinson’s Disease, the article states“For example, PRDX6 accounts for 0.4% of the total ion abundanceafter DOC (deoxycholate) extraction, whereas following HV ( HemoVoid™)extraction, this increases to 8%, a 20-fold enrichment”. Theauthors conclude that the HemoVoid™ method significantly reducesthe concentration of hemoglobin, resulting in an increased signal-tonoise of the remaining red cell proteins.
Elhadi,Suaad Abd, et al. "α”Synucleinin blood cells differentiates Parkinson’s disease from healthycontrols ." Annalsof Clinical and Translational Neurology. Thegoal of this study was todetermine whether blood cells expressing α-Synuclein candifferentiate Parkinson’s disease (PD) from healthy controls. Two proteoforms- PSer129 a-Syn( phosphorylatedpathological form in Lewy bodies) andOxidized a-Synlevels are observed in blood cells, but both at considerably lowerconcentration than total a-Syn,so the extremely high abundance of hemoglobin interferes with theiranalysis. To compensate, the article states for PSer129 α -Syn &Oxidized α -Syn detection by immunoassay, “ followedfrom hemoglobin clearance with HemoVoid kit”.
VicenteMiranda, Hugo, et al. " Posttranslationalmodifications of blood-derived alpha-synuclein as biochemical markersfor Parkinson’s disease ." Scientificreports 7.1(2017): 13713.
Posttranslational modifications (PTMs) in aSyn have beenidentified and implicated on its pathobiology. Since aSyn is abundantin blood erythrocytes, the study aimed to evaluate whether PTMs ofaSyn in the blood might hold value as a biomarker for PD. Ashaemoglobin is the major protein component of erythrocytes lysates(90%), the article states “we depleted this … using HemoVoid…, enabling the additional concentration of otherproteins of lower abundance. We confirmed the aSyn enrichment by immunoblotting (SDS-PAGE and dot-blot).”
RedBlood Cells (RBCs) / Other Applications
Mitra,Nibedita, et al. "Multi-omicsanalysis of red blood cells reveals thalassemia severity beyondglobin gene mutations." BloodAdvances (2026):bloodadvances-2025016677.
HemoVoid™enrichment was used in a multi-omic study of thalassemia severity.
Mitra,Nibedita, et al. " Multi-OmicsAnalysis of Red Blood Cells Reveals Molecular Pathways UnderlyingThalassemia Severity Beyond Globin Gene Mutations ." medRxiv (2025):2025-02.
Thestudy aims were to identify dysregulated molecular pathways in redblood cells contributing to thalassemia severity. In the methodssection for Sample Preparation for RBC Proteomics Study, the articlestates “hemoglobin was depleted using the HemoVoid kit…”. This investigation findssix pathways which are responsible for thalassemia severityindependent of mutational burden.
Wu,Na, et al. " Proteomiccharacteristics of plasma and blood cells in natural aging rhesusmonkeys ." Proteomics:2200049. This study sought to understand the aging process. For thispurpose, the investigation analyzed and compared the proteinexpression spectrums in the blood of old and young rhesus monkeys. Toextract blood cell proteins and deplete Hemoglobin, the articlestates “Blood cell proteins were lysed with…protein extractionsolution (Bestbio, China)…After centrifugation…the supernatantswere further depletedof Hemoglobin using HemoVoid™ .”Upon depletion, the study found 1183 proteins expresseddifferentially in blood cells.
Das,Sonu, et al. " Ajourney to unravel the pathophysiology of stable and exacerbatedChronic obstructive pulmonary disease through erythrocyte proteomics:A combined mass spectrometry/bioinformatics approach ."(2022). A label free relative quantification of erythrocyte cytosolproteome based on LC-MS/MS was performed on hemoglobin- depletederythrocyte lysate samples of stable and exacerbated COPD, relativeto healthy controls. To deplete Hemoglobin, the article states“ HemoVoid™,… from erythrocyte lysate samples to unmask low abundance…proteins….”
Pawliński,”ukasz, et al. " Proteomicbiomarkers in Gaucher disease ." Journalof clinical pathology 74.1(2021): 25-29. For proteomics, quantitativeanalysis was performed by the isobaric tag for a relative andabsolute quantitation (iTRAQ) method. The article states “Cellswere lysed in lysis buffer (7 mol/Lurea, 2 mol/Lthiourea, 4% CHAPS, 1% DTT …,vortexed, incubated at 25 °Cfor 30 minand then centrifuged at 12 000 ×gfor 15 min…thesamples were purified using HemoVoid resin (Biotech Support Group) to remove haemoglobin contamination.The study found 31 proteins that significantly differed inconcentration between GDt1 patients and a control group.
Christer,M. A. L. M., et al. " Methodsfor the detection of autologous blood-doping. " U.S.Patent Application No. 16/976,936. Thepatent application relates to the detection of autologous blooddoping. More specifically, the invention relates to methodscomprising tryptic digestion of samples of isolated red blood cell(RBC), specifically isolated RBC cytosol, followed by peptide mappingusing LC-MS/MS. The invention’s description states “ Hemoglobindepletion was performed using HemoVoid resin ….”. Upon the preferable depletion of hemoglobin, the methods enabledetection of increased levels of certain peptides in samples fromsubjects that have been subjected to autologous blood doping,compared to samples from non-doped control subjects.
Bollenbach,Alexander, et al. " GC-MSand LC-MS/MS pilot studies on the guanidine (NG)-dimethylation innative, asymmetrically and symmetrically NG-dimethylatedarginine-vasopressin peptides and proteins in human red bloodcells. " Journalof Chromatography B (2020):122024.
Previousstudies showed that human red blood cells are rich in large (> 50kDa) asymmetric dimethylarginine -containing proteins of unknownidentity. The study aimed to report the identity, biological activityand concentration of NG-methylatedproteins by using GC-MS and LCMS/MS approaches. The article states“ we included in our method the useof HemoVoid™ to remove specifically most erythrocytic hemoglobinand to improve the SDS-PAGE separation of proteins for furtherprocessing. The HemoVoid™, … allowed removal of erythrocytichemoglobin to a large extent from the hemolysate .… removal of hemoglobin by this technique enabled an effectiveseparation by SDS-PAGE and isolation of bands… .”.
Kitao,Akihito, et al. " Band3 ectopic expression in colorectal cancer induces an increase inerythrocyte membrane-bound IgG and may cause immune-relatedanemia ." InternationalJournal of Hematology (2020):1-10. Autoimmunehemolytic anemia (AIHA) is a rare comorbidity in colorectal cancer(CRC) and has an unknown etiology. To better understandcancer-related anemia, the authors’ investigated ectopic band 3expression and erythrocyte membrane-bound IgG in a CRC cohort. Toreduce the interference from Hemoglobin, the article states“Erythrocytes were lysed … and hemoglobinwas depleted using HemoVoid ”.
Rosin-Arbesfeld,Rina, and Ronen SIMAN-TOV. "Articleof manufacture and methods for increasing survival of red bloodcells." U.S.Patent Application No. 15/739,857. The patent application describesan ex - vivo method of increasing survival of red blood cells (RBCs).The invention’s description states “The Haemolysates wereenriched with over 95 % hemoglobin.For hemoglobindepletion ,the hemoglobin depletion kit of HemoVoid … was used ”.Upon depletion of hemoglobin, a reduction in cytoplasmic actin levelswas observed.
Nemkov,Travis, et al. " Hypoxiamodulates the purine salvage pathway and decreases red blood cell andsupernatant levels of hypoxanthine during refrigeratedstorage ." haematologica 103.2(2018): 361-372. Thegoal of this study was to use proteomics in part to understandhypoxanthine catabolism invivo forstored red blood cells. It is still unclear whether accumulation ofhypoxanthine in stored red blood cell units is clinically relevantfor transfused recipients. Thearticle states “ Leukocyte-reducedhuman RBC from healthy donor volunteers were washed five times inphosphate-buffered saline prior to lysis in distilled water withsonication. Proteomic analyses of RBC membranes and cytosols wereperformed… RBCcytosolic proteins were depleted of hemoglobin using Hemovoid™ prior to high-pH reversed phase fractionation”.
Cortese-Krott,Miriam M., et al. "Identificationof a soluble guanylate cyclase in RBCs: preserved activity inpatients with coronary artery disease." RedoxBiology (2017).
http://www.sciencedirect.com/science/article/pii/S2213231717306535
Inbrief, the authors aimed to investigate whether RBCs carry afunctional soluble guanylate cyclase (sGC) signalling pathway and toaddress whether this pathway is compromised in coronary arterydisease. The article states “ Usinga commercial resin (HemoVoid™), which removes hemoglobin… andallows enrichment of soluble cytoplasmic proteins, we established aprocedure that allows fast and reliable preparation ofhemoglobin-free cell lysates from as little as 1-2 ml blood .In those samples, expression and activity of the cGMP-generating sGC,cGMP-hydrolyzing PDE5 and cGMP-transducing PKG was assessed byenzymatic assays and Western blot analysis”.
Feliciano,Amélia, et al. "Evening and morning alterations in ObstructiveSleep Apnea red blood cell proteome." Datain Brief (2017). http://dx.doi.org/10.1016/j.dib.2017.01.005
Usingproteomics-based evaluation of red blood cells (RBC), the authorsidentified differentially abundant proteins associated withObstructive Sleep Apnea Syndrome (OSA). Proteome variations betweenvarious time points were assessed. The article states “ RBCcytoplasmic fraction depleted of hemoglobin, using HemoVoid™ system, were analyzed by two-dimensional fluorescence difference gelelectrophoresis (2D-DIGE), the 2D image software-based analyzed andrelevant differentially abundant proteins identified by massspectrometry (MS)”.
PhilippF Lange, Pitter F Huesgen, Karen Nguyen, and Christopher M Overall.” AnnotatingN termini for the Human Proteome Project: N termini andNα-acetylation status differentiate stable cleaved protein speciesfrom degradation remnants in the human erythrocyte proteome”, J.Proteome Research., Just Accepted Manuscript • DOI:10.1021/pr401191w • 21 Feb 2014. Thearticle describes a goal of the Chromosome-centric Human ProteomeProject to identify all human protein speciesUsing a N-terminomicsprocedure called TAILS, the authors identified from the HemoVoid™treated, soluble fraction, 778 proteins were identified, 171 of whichwere not represented in either the soluble non-depleted fraction orthe membrane fraction.
KatjaWalpurgis, Maxie Kohler, Andreas Thomas et al. Validatedhemoglobin-depletion approach for red blood cell lysate proteomeanalysis by means of 2D-PAGE and Orbitrap MS .Electrophoresis.2012;
Mizukawa,B., George, A., Pushkaran, S. et al. CooperatingG6PD mutations associated with severe neonatal hyperbilirubinemia andcholestasis .Pediatric Blood Cancer.2011;56: 840-842.
SudhaNeelam, David G Kakhniashvili, Stephan Wilkens et al. Functional20S proteasomes in mature human red blood cells ExperimentalBiology and Medicine.2011;236:580-591
RedBlood Cells, Plasmodium extracts / Malaria
Machado,Patrícia Isabel Pires. Pyruvatekinase and glucose-6-phosphate dehydrogenase deficiencies and theirassociation with malaria–population genetics and proteomic studies . Diss.Universidade do Porto, 2013.
Walpurgis,Katja, et al. " Effectsof gamma irradiation and 15 days of subsequent ex vivo storage on thecytosolic red blood cell proteome analyzed by 2D DIGE and OrbitrapMS ." PROTEOMICS-ClinicalApplications (2013).
LasonderE, Green JL, Camarda G, Talabani H, Holder AA, Langsley G, AlanoP. ThePlasmodium falciparum schizont phospho-proteome reveals extensivephosphatidylinositol and cAMP-Protein Kinase A signalling .J Proteome Research. 2012;
SpeciesAgnostic Applications
Lan,Qin, and Zhao-bing Gu. " Data-independentacquisition-based proteome profiling of red blood cells from dairybuffaloes under different types of heat stress ." Veterinaryand Animal Science (2025):100437. Heatstress (HS) induces hypoxia and oxidative stress, reducing animalhealth and livestock production. Red Blood Cell (RBC) lysates wereisolated for data-independent acquisition-based proteomics toidentify differentially expressed proteins involved in the HSresponse. The article states “ HemoVoid™LC-MS On-Bead kit (Biotech Support Group, HVB-MS10) was used to remove Hb”.Digested peptides were analyzed by nanometric high performance liquidchromatography performed with a Q-Exactive HFX (ThermoScientific) mass spectrometer, operated in DIA mode. Results showedthat blood clotting factors, complement proteins, immunoglobulins,and vasoconstriction proteins were consistently decreased under thethree types of HS conditions.
Puente-Marin,Sara, et al. "InSilico Functional Networks Identified in Fish Nucleated Red BloodCells by Means of Transcriptomic and Proteomic Profiling ." Genes 9.4(2018): 202. Label-freeshotgun proteomic analyses were carried out for in silico functionalpathway profiling of rainbow trout RBCs. Thearticle states “ Thecytosolic fraction, approximately 300 μL, was depleted of hemoglobinusing HemoVoidTM kit(Biotech Support Group, Monmouth Junction, NJ, USA), in accordancewith the manufacturer’s instructions” .
NombelaI, Puente-Marin S, Chico V etal. Identificationof diverse defense mechanisms in trout red blood cells in response toVHSV halted viral replication F1000Research 2017, 6:1958(doi: 10.12688/f1000research.12985.1)
Thearticle states “ …LC ESI-MS/MS analysis of each of the fractions… . Briefly, thehaemoglobin of the cytosolic fraction was removed using a column of HemoVoid™ kit…,following the manufacturer instructions ”.
Fora full list of Hemoglobin Removal references, visit:
https://www.biotechsupportgroup.com/References-s/138.htm#hemoglobin-depletion
