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Home > Antibodies > Anti-Human Antibodies for Flow Cytometry > Anti-human CD64 Monoclonal Antibodies for Flow Cytometry

Anti-human CD64 Monoclonal Antibodies for Flow Cytometry

cluster of differentiation 64, Fc gamma RIA and Fc gamma RIB, Human Fc Receptor Blocking Antibody

Catalog No. Product Name Size List Price (US$) Quantity
PA007420.m1 Recombinant Anti-human CD64 Monoclonal Antibody (Clone: 22), Mouse IgG1 Kappa 100 ug 50.00
PA007420.m1 Recombinant Anti-human CD64 Monoclonal Antibody (Clone: 22), Mouse IgG1 Kappa 500 ug 100.00
PA007420.h1Fs Recombinant Anti-human CD64 Monoclonal Antibody (Clone: 22), Human IgG1 Fc Silent Kappa 100 ug 50.00
PA007420.h1Fs Recombinant Anti-human CD64 Monoclonal Antibody (Clone: 22), Human IgG1 Fc Silent Kappa 500 ug 100.00
Description

PA007420.m1: Non-conjugated Recombinant Anti-human CD64 Monoclonal Antibody (Clone: 22), Mouse IgG1 Kappa

Recombinant  Mouse IgG1 Monoclonal Antibody. 
Clone: 22.
Isotype:  Mouse IgG1 kappa.
Source: The anti-human CD64 monoclonal antibody (clone: 22) was produced in mammalian cells.
Conjugation: Non-conjugated.
Specificity/Sensitivity: The recombinant 22 antibody specifically binds to human CD64.
Applications: Flow cytometry (FC), and Immunohistochemistry - Frozen (IHC-F). 
Form of Antibody: 0.2 uM filtered solution, pH 7.4, containing 0.09% sodium azide.
Purity: >95% by SDS-PAGE under reducing conditions.

PA007420.h1Fs: Non-conjugated Recombinant Anti-human CD64 Monoclonal Antibody (Clone: 22), Human IgG1 Fc Silent Kappa

Recombinant  Human IgG1 Monoclonal Antibody with Fc silent mutation. 
Clone: 22.
Isotype: Human IgG1 kappa.
Source: The anti-human CD64 monoclonal antibody (clone: 22) was produced in mammalian cells.
Conjugation: Non-conjugated.
Specificity/Sensitivity: The recombinant 22 antibody specifically binds to human CD64.
Applications: Flow cytometry (FC), and Immunohistochemistry - Frozen (IHC-F). 
Form of Antibody: 0.2 uM filtered solution, pH 7.4, containing 0.09% sodium azide.
Purity: >95% by SDS-PAGE under reducing conditions.

Shipping: The 22 antibody is shipped with ice pack. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 
12 months from date of receipt, 2 to 8°C as supplied. Do not freeze it.

Syd Labs provides the following recombinant anti-human CD64 monoclonal antibodies for flow cytometry:

Non-conjugated Recombinant Anti-human CD64 Monoclonal Antibody (Clone: 22), Mouse IgG1 Kappa
Non-conjugated Recombinant Anti-human CD64 Monoclonal Antibody (Clone: 22), Human IgG1 Fc Silent Kappa

Background

The recombinant 22 antibody binds to human and non-human primate CD64 or FcγRI, a cluster of differentiation molecule found on monocytes, macrophages, and neutrophils. CD64 plays a central role in macrophage antibody-dependent cellular cytotoxicity and clearance of immune complexes, and is a candidate biomarker for bacterial infection and sepsis.

The most popular human Fc blocking reagents include:
Anti-human CD16 (clone 3G8), and CD32 (Clone IV.3): for flow cytometry, immunohistochemistry (IHC), and immunoprecipitation (IP). It is highly specific for FcγRII (CD32) and FcγRIII (CD16), and reduces background staining; it may not block all Fc receptors. Clones 10.1, and 22 or H22 are the most widely used anti-human FcγRI (CD64) clone for Fc blocking in flow cytometry and functional assays. It prevents non-specific Fc receptor binding on monocytes, macrophages, and dendritic cells, improving antibody specificity.
Purified human IgGs or mixes of human isotype controls: for general Fc blocking in IHC and immunofluorescence (IF). It is cheap and easy to use but it is less specific, and may even introduce unwanted immunoglobulins. Syd Labs supply various human IgG isotype controls.
Normal mouse serum, purified mouse IgG or isotype controls: for flow cytometry. It can serve as a control to block Fc receptors but it is non-specific for Fc receptors. It is cheap and easy to use but it is less specific, and may even introduce unwanted immunoglobulins. Our recombinant mouse IgG antibody mixes are affordable; the ratio of various IgGs is optimized and adjustable without any unwanted immunoglobulins.
Bovine Serum Albumin (BSA) or FBS: for general blocking in Western Blot (WB) and IHC. They are readily available to reduce non-specific binding but less effective at Fc receptor blocking.
Commercial Fc blocking solutions: for flow cytometry, IHC, and functional assays. They are normally pre-optimized and highly effective but more expensive than DIY solutions. 

Best choice of human Fc blocking reagents based on your application are:
For flow cytometry: anti-human CD16 (Clone 3G8), CD32 (Clone IV.3), and CD64 (Clone 10.1, and 22 or H22).
For Immunohistochemistry (IHC) and Immunofluorescence (IF): Normal mouse serum, purified mouse or human IgGs or isotype controls or commercial Fc blocking kits. Our recombinant mouse IgG antibody mixes and recombinant human IgG isotype controls are affordable; the ratio of various IgGs is optimized and adjustable without any unwanted immunoglobulins.
For Western Blot and ELISA: BSA or FBS.
For functional assays (e.g., blocking Fc-mediated effects): Commercial Fc blocking reagents.

References of anti-human CD64 antibody (H22):


Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition)
Cossarizza A, et al. Eur J Immunol. 2019 Oct;49(10):1457-1973. doi: 10.1002/eji.201970107. PMID: 31633216
These guidelines are a consensus work of a considerable number of members of the immunology and flow cytometry community. They provide the theory and key practical aspects of flow cytometry enabling immunologists to avoid the common errors that often undermine immunological data. Notably, there are comprehensive sections of all major immune cell types with helpful Tables detailing phenotypes in murine and human cells. The latest flow cytometry techniques and applications are also described, featuring examples of the data that can be generated and, importantly, how the data can be analysed. By analyzing tissue and blood from the same patients, we have also demonstrated that monoclonal circulating cells detected by flow cytometry reflect peripheral circulating tumor cells.
Tags: H22 antibody; H22 antibody for animal model

Recombinant H22(scFv) blocks CD64 and prevents the capture of anti-TNF monoclonal antibody: A potential strategy to enhance anti-TNF therapy
Hristodorov D, et al. MAbs. 2015;7(1):71-8. doi: 10.4161/mabs.32182. PMID: 25517313
We used a single chain antibody fragment, H22(scFv), which binds specifically to CD64. CD64-selective blocking was facilitated by the heterologous expression of an Fc-free, anti-CD64, single chain antibody fragment, called H22(scFv). The addition of recombinant H22(scFv) to stimulated HL-60 cells resulted in the potent and specific blocking of CD64, thus strongly reducing the ability of the cells to capture anti-TNF mAb. In contrast, CD64 binding of H22(scFv) induced neither receptor activation nor the expression of pro-inflammatory cytokines. In conclusion, the CD64-blocking antibody fragment H22(scFv) prevents the capture of anti-TNF mAb by CD64, whose overexpression on the surface of inflammatory monocytes and macrophages is associated with the poor response of some patients toward anti-TNF therapy.
Tags: H22 antibody for cancer research; H22 antibody for flow cytometry

Validation of a Flow Cytometry Based Binding Assay for Evaluation of Monoclonal Antibody Recognizing EGF Receptor
Cedeño-Arias M, et al. Sci Pharm. 2011 Oct-Dec;79(4):793-809. doi: 10.3797/scipharm.1104-18. PMID: 21886904
The assay was validated by examining, assay robustness, specificity, repeatability and intermediate precision. The assay was highly specific, robust for all studied factors except for cell fixation with 1% paraformaldehyde and met criteria for precision with RSD < 2%. In addition the assay has stability-indicating properties evidenced by the ability to detect changes in mAb degraded samples. Most importantly, the assay demonstrated to be useful for its intended use. Development of a convenient, robust, specific and precise procedure is important in quality control laboratories.
Tags: H22 antibody for human tumor model; H22 antibody human dose range

Flow cytometry with anti HLA-antibodies: a simple but highly sensitive method for monitoring chimerism and minimal residual disease after HLA-mismatched stem cell transplantation
Schumm M, et al. Bone Marrow Transplant. 2007 Jun;39(12):767-73. doi: 10.1038/sj.bmt.1705676. PMID: 17438586
Transplantation of HLA-mismatched stem cells may allow determination of chimerism status of single cells by differential expression of HLA molecules. Monoclonal antibodies against HLA antigens can be used to determine the HLA type of sub-populations by standard flow cytometry. Information about the chimerism status is given on a single-cell level and allows fast and convenient analysis of sub-populations. In addition to chimerism evaluation, HLA antibodies improved the detection of leukemic cells after transplantation with aberrant phenotype. In conclusion, flow cytometry using antibodies against HLA antigens is an interesting tool for determination of chimerism and minimal residual disease after HLA-mismatched transplantation.
Tags: activity of H22 antibody; function of H22 antibody

The Use of Monoclonal Antibodies and Flow Cytometry in the Diagnosis of Paroxysmal Nocturnal Hemoglobinuria
Hall SE, et al. Blood. 1993 Oct 15;82(8):2360-8. PMID: 8365783
Flow cytometry provides a rapid qualitative method for analyzing the binding of a fluorescent probe to a cell. To quantitate the binding of a probe using flow cytometry, one must be able to first calibrate the fluorescent signal with some known standard. We have compared a new one-step method with a previous two-step method for determining the number of binding sites (receptors) on the surface of cells using immunofluorescent staining of the cells and analysis by flow cytometry. Experimentally, recombinant chinese hamster ovary cells, expressing cell surface glycoprotein receptors, IIb/IIIa or Mac-1, were assayed using specific mouse monoclonal antibodies against these receptors. The assay was highly specific, robust for all studied factors except for cell fixation with 1% paraformaldehyde and met criteria for precision with RSD < 2%.
Tags: H22 mAb for cancer research; H22 monoclonal antibody

For more references about anti-human CD64 antibody (H22), please contact our scientific support team with message@sydlabs.com.

Syd Labs provides the following in vivo grade recombinant anti-human CD16, CD32, and CD64 monoclonal antibodies:
Recombinant Anti-human CD16 monoclonal antibody (Clone: 3G8)
Recombinant Anti-human CD32 monoclonal antibody (Clone: IV.3)
Recombinant Anti-human CD64 monoclonal antibody (Clone: H22)

Syd Labs provides the following in vivo grade recombinant anti-mouse CD16, CD32, and CD64 monoclonal antibodies:
Recombinant Anti-mouse CD16/CD32 monoclonal antibody (Clone: 2.4G2)

Syd Labs provides the following recombinant anti-human CD16, CD32, and CD64 monoclonal antibodies for flow cytometry:
Recombinant Anti-human CD16 monoclonal antibody (Clone: 3G8) for flow cytometry
Recombinant Anti-human CD32 monoclonal antibody (Clone: IV.3) for flow cytometry
Recombinant Anti-human CD64 monoclonal antibody (Clone: H22) for flow cytometry

If you need to learn more about Anti-human CD64 Monoclonal Antibodies for Flow Cytometry, click: Anti-human CD64 Antibody (H22) | PA007393.m1

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