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HemogloBind™ Hemoglobin Depletion From Hemolyzed Serum/Plasma

HemogloBind™ Hemoglobin Depletion From Hemolyzed Serum/Plasma


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Product Code: H0145

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Hemoglobin Removal and Capture

  • Has a high degree of specificity for hemoglobin, without cross-reacting with other proteins or analytes

  • Suitable for

    • Hemolyzed serum/plasma

    • Whole Blood, Dried Blood Cards (DBS) or Erythrocyte lysates

    • Tissue Homogenates, compatible with RIPA buffer

  • Applications in analytical interferences, enzyme monitoring, proteomics

  • Species Agnostic

  • Over 30 citations including analysis in cellular thermal shift assay (CETSA), LC-MS proteomics, Hemoglobin derivatives, Western blot, enzyme activity, ELISA

  • Related product NuGel™HemogloBind™ comes in a dry powder format, compatible with high throughput 96-well automation, see https://www.biotechsupportgroup.com/NuGel-HemogloBind-Hemoglobin-Capture-Reagent-p/np-ho.htm

Poly-electrolytes are polymers with repeating units of stationary charges. HemogloBind™ is derived from insoluble elastomeric poly-electrolytes that bind proteins through an empirically derived chemistry combining elements of polymer composition, and cross-linking architecture. As with bio-polymers like DNA and Heparin, governing their reactivity is the spatial presentation of electrostatic groups along a flexible polymer chain.

HemogloBind™ does not cross react with most common serum components, making it an excellent tool in numerous applications. These include analytical protocols where optical interference is problematic. Hemoglobin variants, as in thalassemia, bind with differential affinity towards HemogloBind™, though this has not been fully evaluated. For purification and/or analysis of hemoglobin, a modest elevation in pH will facilitate desorption from the polymer.

Click Here To View HemogloBind™ Product Sheet
Click Here For The HemogloBind™ Hemoglobin Depletion Performance On Blood Sheet
Click Here For The HemogloBind™ Hemoglobin Depletion Performance On Dried Blood Spots/Cards

Hemolyzed Samples

Neil Adrian P. Ondevilla, Peng-Wen Liu, Wan-Ting Huang, Tzu-Ping Weng, Nan-Yao Lee, Syu-Cing Ma, Jian-Jang Huang, Tak-Wah Wong, Hsien-Chang Chang, A point-of-care electrochemical biosensor for the rapid and sensitive detection of biomarkers in murine models with LPS-induced sepsis, Biosensors and Bioelectronics, Volume 254, 2024, 116202, ISSN 0956-5663, https://doi.org/10.1016/j.bios.2024.116202.

Pooled Blood Samples

Igoh, Akihisa, Masanobu Miura, and Satoru Miyaishi. "Animal Species and Blood Identification with Peptide Mass Fingerprinting." Analytical Chemistry (2024).

RBC lysates

Chen, Yaozhen, et al. "Noncanonical NLRP3 Inflammasome Activation Elicits the Programmed Death of Red Blood Cells ." Available at SSRN 4626203 .

Abdominal Aneurysm Samples

Jara, Zaira Palomino, et al. "Distinct Mechanisms of β-Arrestin–Biased Agonist and Blocker of AT1R in Preventing Aortic Aneurysm and Associated Mortality ." Hypertension 80.2 (2023): 385-402.

Peripheral blood mononuclear cells (PBMC) whole cell lysate samples

Kaneko, Tomonori, et al. " Kinome and phosphoproteome reprogramming underlies the aberrant immune responses in critically ill COVID-19 patients ." (2023).

Red blood cells

Barakat, Amey, et al. " Effects of 2, 3‐DPG knockout on SCD phenotype in Townes SCD model mice. " American Journal of Hematology (2023).

Mouse Red Blood Cells
Zhan, Jing, et al. "
Silica nanoparticles trigger phosphatidylserine exposure in red blood cells and induce thrombosis risk ." Environmental Pollution (2023): 121591.


Dasauni, Pushpanjali, et al. "Optimization and Identification of Single Mutation in Hemoglobin Variants with 2, 2, 2 Trifluoroethanol Modified Digestion Method and Nano− LC Coupled MALDI MS/MS ." Molecules 27.19 (2022): 6357.

hole Blood Sample

Witchey, Shannah K., et al. "Reproductive and developmental toxicity following exposure to organophosphate ester flame retardants and plasticizers, triphenyl phosphate and isopropylated phenyl phosphate, in Sprague Dawley rats." Toxicological Sciences (2022). https://doi.org/10.1093/toxsci/kfac135

emolysate Preparations Remnant White Ghosts
Sae-Lee, Wisath, et al. " The protein organization of a red blood cell ." Cell Reports 40.3 (2022): 111103.

a Samples

Phillippi, Danielle T., et al. " Inhaled diesel exhaust particles result in microbiome-related systemic inflammation and altered cardiovascular disease biomarkers in C57Bl/6 male mice ." Particle and fibre toxicology 19.1 (2022): 1-29.

Urine Samples

Juriasingani, Smriti, et al. " Evaluating the Effects of Subnormothermic Perfusion with AP39 in a Novel Blood-Free Model of Ex Vivo Kidney Preservation and Reperfusion. " International Journal of Molecular Sciences 22.13 (2021): 7180.

Lysed RBCs

D'Alessandro, Angelo, et al. " Hematologic and systemic metabolic alterations due to Mediterranean type II G6PD deficiency in a novel murine model. " bioRxiv (2021).

Hemolyzed Red Blood Cells Biological Fluids

Heather E. McKiernan, Phillip B. Danielson, Catherine O. Brown, Masha Signaevsky, Christian G. Westring and Kevin M. Legg, Developmental Validation of a Multiplex Proteomic Assay for the Identification of Forensically Relevant Biological Fluids, Forensic Science International, (2021) https://www.sciencedirect.com/science/article/pii/S0379073821002280?via%3Dihub

Peripheral blood derived mononuclear cells whole cell lysate samples

Kaneko, Tomonori, et al. " System-wide hematopoietic and immune signaling aberrations in COVID-19 revealed by deep proteome and phosphoproteome analysis. " Research Square preprint (2021).

Blood cell lysates

Yamagishi, Yoshikazu, Hirotaro Iwase, and Yasumitsu Ogra. "Post-Mortem Changes of Methomyl in Blood with Hemoglobin." Chemical Research in Toxicology.

Reticulocytes (Retics) lysates

Hojo-Souza NS, de Azevedo PO, de Castro JT, Teixeira-Carvalho A, Lieberman J, et al. (2020) Contributions of IFN-γ and granulysin to the clearance of Plasmodium yoelii blood stage. PLOS Pathogens 16(9): e1008840. https://doi.org/10.1371/journal.ppat.1008840


Southwell, Rebecca Marie, Kenneth Sherlock, and Matthew Baylis. "Cross-sectional study of British wild deer for evidence of Schmallenberg virus infection. " Veterinary Record (2020).

Host red blood cells

Dziekan, Jerzy Michal, et al. "Cellular thermal shift assay for the identification of drug–target interactions in the Plasmodium falciparum proteome. " Nature Protocols (2020): 1-41.

Featured HemogloBind ™ Reference Applications –

Nguyen, Anthony T., et al. " UBE2O remodels the proteome during terminal erythroid differentiation. " Science 357.6350 (2017): eaan0218.

During reticulocyte maturation, the proteome is remodeled through the programmed elimination of most generic constituents of the cell, in parallel with abundant synthesis of hemoglobin. The study used multiplexed quantitative proteomics to identify candidate substrates of UBE2O, an E2 (ubiquitin-conjugating) enzyme, in an unbiased and global manner. Because of the overly abundant presence of Hemoglobin, selective depletion of Hemoglobin was necessary. T he article states “Reticulocytes were lysed by vortexing for 5 minutes at room temperature… An additional 10 bed vol of Hemoglobind™ suspension was added to the samples, which were then vortexed for another 10 min at room temperature followed by 4 min of centrifugation at 10000 x g. The supernatants, which contain hemoglobin-depleted sample, were … processed for TMT quantification. ”.

Hemolyzed Serum Exosome Analyses

Nishida-Aoki, Nao, et al. "Disruption of Circulating Extracellular Vesicles as a Novel Therapeutic Strategy against Cancer Metastasis." Molecular Therapy 25.1 (2017): 181-191.


This study considers that therapeutic strategies targeting cancer-derived extracellular vesicles (EVs) hold great promise because of the possibility they reposition microenvironments to accommodate metastasis. The researchers report on a novel strategy of therapeutic antibody treatment to target cancer-derived EVs and inhibit the metastasis of breast cancer in a mouse model. The article states “Hemoglobin was accumulated with HemogloBind™ beads (Biotech support group, Monmouth Junction NJ, USA) followed by 0.22 μm filtration. Then, the EVs in the sera were concentrated by ultracentrifugation…”.

Macromolecular Complexes
and Functional Integrity

C Wan, B Borgeson, S Phanse, F Tu, K Drew, G Clark, et al. Panorama of ancient metazoan macromolecular complexes. Nature Volume:525, Pages:339–344 Date published:(17 September 2015). doi:10.1038/nature14877

HemogloBind™, contributed to this rigorous examination of protein complexes. When our products were used as a pretreatment step in the overall workflow, about twice the number of observations and annotations became possible. Furthermore, this study demonstrated the importance of a key feature implicit to all of our products; that is the maintenance of functional and structural integrity after separations. Without that particular feature, these additional observations would not have been possible.

Whole Blood

Lahut, Suna, et al. "Blood RNA biomarkers in prodromal PARK4 and REM sleep behavior disorder show role of complexin-1 loss for risk of Parkinson's disease." Disease Models & Mechanisms (2017): dmm-028035. http://dmm.biologists.org/lookup/doi/10.1242/dmm.028035

In this study, Parkinson’s disease progression is investigated through the accumulation and aggregation of the lipid-binding SNARE complex component alpha-synuclein (SNCA) which underlies vulnerability and defines its stages. The authors studied blood samples from a new large pedigree with SNCA gene duplication (PARK4 mutation), to identify effects of SNCA gain-of-function as potential disease biomarkers. The article states “For protein extraction from the EDTA tubes, 300 μl blood were lysed with equal amount of 1% SDS-RIPA buffer [50 mM Tris-HCl (pH 8.0), 150 mM NaCl, 1 mM EDTA, 1 mM EGTA, 1% Igepal CA-630 (Sigma), 0.5% sodium deoxycholate, 0.1% SDS, 1 mM PMSF and one tablet Complete Protease Inhibitor Cocktail (Roche)] and sonicated for 10 sec. The blood lysates were rotated at 4 °C for 30 min and centrifuged at 4 °C for 30 min. The supernatants were depleted in hemoglobin content using a commercial kit (HemogloBind™) following the manufacturer’s instructions”.

Chalásová, Katarína, et al. "Transketolase Activity but not Thiamine Membrane Transport Change in Response to Hyperglycaemia and Kidney Dysfunction." Experimental and Clinical Endocrinology & Diabetes (2017).

Diabetic kidney disease, a common complication of both type 1 and type 2 diabetes, is associated with significant morbidity and mortality, and represents the most common cause of chronic kidney disease. The study hypothesized that protective pentose phosphate pathway action in diabetes might be compromised by limited intracellular availability of an active transketolase cofactor thiamine diphosphate (TDP). To evaluate the levels of thiamine tranporter proteins in whole blood, t he article states “ For protein isolation, whole blood aliquots were lysed with water and haemoglobin was removed using HemogloBind™ (Biotech Support Group) according to manufacturer’s instructions…”.

Species Agnostic – Applications to Different Species

Snider, Thomas H., Christina M. Wilhelm, Michael C. Babin, Gennady E. Platoff Jr, and David T. Yeung. " Assessing the therapeutic efficacy of oxime therapies against percutaneous organophosphorus pesticide and nerve agent challenges in the Hartley guinea pig ."The Journal of Toxicological Sciences 40, no. 6 (2015): 759-775.

Acetylcholine is an essential neurotransmitter, and inhibitors of cholinesterases(ChEs) are potent toxins. A primary component of anti-organophosphorus therapy is an oxime reactivator to rescue inhibited acetylcholinesterases. For this, clinical signs of toxicity can be measured from blood cholinesterase [Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE)] activity utilizing a modified Ellman's method. Biotech Support Group’s unique solid-phase polymer for hemoglobin depletion, was used for pretreatment. The article states “Briefly, whole blood samples were treated with HemogloBind™ which interferes with the ChE activity assay due to spectral overlap.”

Craig, J. R., et al. "A comparison of the anatomical and gastrointestinal functional development between gilt and sow progeny around birth and weaning ." Journal of animal science (2019).

Gilt progeny (GP) often have restricted growth performance and health status in comparison to sow progeny (SP) from birth. To better understand underlying mechanisms, the study aimed to compare differences in growth and development between GP and SP in the first 24 h after birth and in the peri-weaning period. Because serum samples were quite hemolysed after collection and processing, it became necessary to use HemogloBind™ to allow for better detection of IgG by ELISA. The article states “As per the manufacturer’s instructions, 250 μL of Hemoglobind was added to 250 μL of hemolyzed serum…

Parvathi S. Kumar, Haree K. Pallera, Pamela S. Hair, Magdielis Gregory Rivera, Tushar A. Shah, Alice L. Werner,Frank A. Lattanzio, Kenji M. Cunnion, and Neel K. Krishna. Peptide inhibitor of complement C1 modulates acute intravascular hemolysis of mismatched red blood cells in rats. TRANSFUSION Volume 00, May 2016. doi:10.1111/trf.13674.

In brief, the study evaluated the role of a peptide inhibitor of complement C1 (PIC1) in an animal model of acute intravascular hemolysis in both prevention and rescue scenarios. The authors state "To remove free Hb that may cause optical interference in bilirubin analysis, we treated all the samples with Hb depletion from hemolyzed serum/plasma (HemogloBind, Biotech Support Group). Bilirubin concentration was then measured with a Bilirubin Assay Kit (Sigma-Aldrich, St. Louis, MO)."

Other References

Blood Substitutes

Laing, Richard W., et al. " The use of an acellular oxygen carrier in a human liver model of normothermic machine perfusion ." Transplantation 101.11 (2017): 2746.

Red cell lysates

O’Connell, Grant C., et al. " Monocyte-lymphocyte cross-communication via soluble CD163 directly links innate immune system activation and adaptive immune system suppression following ischemic stroke ." Scientific reports 7.1 (2017): 12940

Kyoungsook Park, Christopher D. Saudek, and Gerald W. Hart Increased Expression of β-N-Acetylglucosamindase (O-GlcNAcase) in Erythrocytes from Prediabetic and Diabetic Individuals. Diabetes.2010;59(7):1845-50.

Delobel J., Rubin O., Prudent M., Crettaz D., Tissot J.-D., Lion N.(2010)
Biomarker Analysis of Stored Blood Products: Emphasis on Pre-Analytical Issues . International Journal of Molecular Sciences. 11(11):4601-4617

Alvarez-Llamas, Gloria, Fernando de la Cuesta, Maria G. Barderas, Irene Zubiri, Maria Posada-Ayala, and Fernando Vivanco. " Characterization of Membrane and Cytosolic Proteins of Erythrocytes. " In Vascular Proteomics, pp. 71-80. Humana Press, 2013.

Alvarez-Llamas, G., de la Cuesta, F., Barderas, M. G., Darde, V. M., Zubiri, I., Caramelo, C., Vivanco, F.
A novel methodology for the analysis of membrane and cytosolic sub-proteomes of erythrocytes by 2-DE. Electrophoresis.2009;30:4095-4108

Zihao Wang, Kyoungsook Park, Frank Comer1, Linda C. Hsieh-Wilson, Christopher D. Saudek, Gerald W. Hart. Site-Specific GlcNAcylation of Human Erythrocyte Proteins: Potential Biomarker(s) for Diabetes Mellitus . Diabetes.2008;58, 309-317.

Yuichi Miki, Tomoki Tazawa, Kazuya Hirano, Hideki Matsushima, Shoko Kumamoto, Naotaka Hamasaki, Tomohiro Yamaguchi, Masatoshi Beppu. Clearance of oxidized erythrocytes by macrophages: Involvement of caspases in the generation of clearance signal at band 3 glycoprotein . Biochemical and Biophysical Research Communications.2007; 363(1):57-62

Sarawathi,et al., Relative quantification of glycated Cu-Zn superoxide dismutase in erythrocytes by electrospray ionization mass spectrometry , Biochimica et Biophysica Acta. 1999.1426(3):483-90