Catalog No.	PA007409
Product Name	Human IgG Antibody Mix with more hIgG1 (Clone: HyHEL-10) | PA007409
Supplier Name	Syd Labs, Inc.
Brand Name	Syd Labs
Isotype	human IgG1, IgG2, IgG3, and IgG4 (S228P) kappa.
Specificity/Sensitivity	neither human nor mouse samples.
Applications	ELISA, flow cytometry, immunoblotting, immunoprecipitation, immunocytochemistry, immunofluorescence, and isotype controls.
Reactivity	None of Human, Mouse, and Rat.
Purification	The recombinant human IgG antibodies were affinity-purified from supernatant of mammalian cells cultured in a chemically defined medium by affinity-chromatography using protein A and G.
Form Of Antibody	1x PBS.
Purity	>95% by SDS-PAGE under reducing conditions.
Shipping	The Recombinant Human IgG Antibody Mix with more hIgG1 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. For maximum recovery of the product, centrifuge the original vial after thawing and before removing the cap. 1 month from date of receipt, 2 to 8°C as supplied. 12 months from date of receipt, -20°C to -70°C as supplied.
Note	Recombinant human IgG antibody mixes: animal free production; lot-to-lot consistency; no binding to human tissues; adjustable formulation of the human IgG antibodies in the antibody mixes. Best normal human serum alternative; normal human serum blocking reagent alternative; serum-derived human IgGs alternative.
Order Offline	Phone: 1-617-401-8149 Fax: 1-617-606-5022 Email: message@sydlabs.com Or leave a message with a formal purchase order (PO) Or credit card.

Description

PA007409: Recombinant Human IgG Antibody Mix with more hIgG1 (Clone: HyHEL-10)

The recombinant human IgG antibody mix is a mixture of recombinant human IgG1, IgG2, IgG3, and IgG4 (S228P) kappa antibodies which bind to neither human nor mouse samples. The percentage of human IgG1 is much higher than the other antibody subclasses in the antibody mix.

Background of Recombinant Human IgG Antibody Mix with more hIgG1

Normal human serum and purified human IgGs from normal human serum are commonly used as non-specific passive blocking solution for antibody-based immuno-assays. Recombinant human IgG antibody mixes, as a direct alternative to normal human serum and even human IgGs purified from human serum or a non-specific passive blocking solution, can be used to eliminate interfering antibodies from immuno-assay systems using capture and detection antibodies. There is higher percentage of human IgG1 in serum. So, recombinant human IgG antibody mixes with different percentages of human IgG1, IgG2, IgG3, and IgG4 (S228P) were made to meet customers’ requirements of different applications. A customized recombinant human IgG antibody mix with different percentages of human IgG1, IgG2, IgG3, and IgG4 can be provided.

Features of recombinant human IgG antibody mixes compared with normal human serum and serum-derived human IgGs:

1) Human free production. CHO cells rather than human are used in antibody production.

2) Lot-to-lot consistency. CHO cells which are used for antibody drug manufacturing are used to manufacture the human IgG antibodies in the antibody mixes. All production is under control and predictable.

3) As a targeted or passive antibody blocker to interfering antibodies and no binding to human tissues. In contrast, some serum-derived human IgGs may bind to unknown targets in some human tissues.

4) Adjustable percentages of the antibodies in the antibody mixes. A customized recombinant human IgG antibody mix with different percentages of human IgG1, IgG2, IgG3, and IgG4 (S228P) can be easily made according to customers’ applications.

Recombinant human IgG antibody mixes are the best normal human serum alternative as blocking reagents, and serum-derived human IgGs alternative.

Questions and Answers:
Question: Normal human serum is often used for blocking or saturating immunoassays, especially those involving tissue samples such as immunohistochemistry (IHC). How much should recombinant antibody mixes be added to replace normal human serum?
Answer: Our recombinant human IgG antibody mixes can be used as normal human serum alternative for blocking reagent, and more specifically, the purified normal IgGs alternative. It was said that there is 65 to 85 mg/ml protein in a normal human serum sample, and 7 to 16 mg/ml IgG in a typical human serum and plasma sample. It means that there is 8.2% to 10.8% IgGs in proteins in normal mouse serum. One may calculate the amount of human IgGs in a human serum sample. One should also consider the percentage of various human IgG isotypes in the human serum sample vs recombinant human IgG antibody mixes. Conditions of optimal human IgG antibody mix performance should be determined experimentally by customers.

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 Recombinant Human IgG Antibody Mixes:

1、Factors Associated With Human IgG Antibody Response to Anopheles albimanus Salivary Gland Extract, Artibonite Department, Haiti, 2017.
Jaramillo-Underwood, A., et al. J Infect Dis. 2022 Oct 17;226(8):1461-1469. doi: 10.1093/infdis/jiac245. PMID: 35711005.
“This study examined the relationship between human IgG responses to a mixture of proteins from the An. …Although the inclusion of the crude salivary gland homogenate in this study ensures numerous proteins are available for IgG binding, it is also not possible to specify from this current study which specific mosquito proteins are capturing the human IgG. …This study’s results suggested that higher elevation and increased distance from water bodies are associated with lower levels of anti-SGE IgG. …Further investigation of these findings is needed using individual participant-level IgG data rather than aggregated mean IgG values, and additionally covariates collected at the individual level that may further explain the relationship between environmental factors and mosquito exposure. …This is in line with this current study’s results as well as those of other groups, which have not observed consistently significant associations between antisalivary IgG and IgG against PfMSP-1.”

2、IgG antibody responses to Anopheles gambiae gSG6-P1 salivary peptide are induced in human populations exposed to secondary malaria vectors in forest areas in Cameroon.
Ndo, C., et al. PLoS One. 2022 Nov 10;17(11):e0276991. doi: 10.1371/journal.pone.0276991. PMID: 36355922.
“Recently, human IgG antibodies specific response to one salivary peptide from the An. …This study was carried out during a pre-intervention survey as part of a pilot project aiming to assess the impact of house improvement in reducing the exposure of human populations to the bites of malaria vectors in the locality of Nyabessang (2°24’ North and 10°24’ East), one of the sentinel sites for malaria entomological surveillance in Cameroon. …The populations of the study site were sensitized a week earlier and invited to participate in the study. …The entomological survey (Anopheles mosquito density and species composition) was assessed in October 2019 at the pick of the rainy season and two weeks before the parasitological survey. …Human IgG antibody response to Anopheles gambiae gSG6-P1 salivary peptide was reported to be a pertinent indicator for assessing human exposure to mosquito bites and evaluating the risk of malaria transmission as well as the effectiveness of vector control strategies.”

3、Human papillomavirus type 16 (HPV-16) IgG antibody among women of reproductive age presenting at a healthcare facility in Central Nigeria: a pilot study.
Adoga MP, et al. Pan Afr Med J. 2021 Dec 6;40:203. doi: 10.11604/pamj.2021.40.203.28804. PMID: 35136466.
“This was followed by adding horseradish peroxidase (HRP)-conjugated anti-human IgG and a 30-minute incubation at 37°C. It was then washed thoroughly. …A microplate reader (Tecan Infinite m200 Pro, Tecan Group Ltd., Mannedorf) was used at 450 nm to spectrometrically measure the intensity of colour changes. …In addition, forty-six (11.5%) reported having the history of genital warts with 268 (67.0%) and 132 (33.0%) subjects being married and single respectively. …In this study, we found high serological evidence of exposure to HPV-16 to be 32.0% by determining the proportion of individuals who had HPV-16 specific IgG antibody among women of reproductive age who presented at a healthcare facility in Central Nigeria. …Additionally, exposure to the virus was not significantly associated with marital status even though evidence of exposure to the virus was slightly higher among individuals who were single with 36.4% seroprevalence than the married subjects who had a seroprevalence of 29.9%.”

4、Human IgG Antibody Response to Aedes Nterm-34kDa Salivary Peptide, an Epidemiological Tool to Assess Vector Control in Chikungunya and Dengue Transmission Area.
Elanga Ndille, E., et al. PLoS Negl Trop Dis. 2016 Dec 1;10(12):e0005109. doi: 10.1371/journal.pntd.0005109. PMID: 27906987.
“The present study aimed to investigate whether human IgG antibody specific response to only one Aedes salivary peptide could be useful for assessing the efficacy of vector control against arboviral diseases. …For this purpose, IgG response to Nterm-34kDa peptide was assessed from 102 individuals living in urban area at La Reunion Island, Indian Ocean, before and after the implementation of vector control against Aedes albopictus mosquito species. …The results of the present study indicated that human IgG response to the Aedes Nterm-34kDa salivary peptide could be a useful tool to evaluate the efficacy of vector control strategies against arboviruses. …For this purpose, human IgG responses to the Nterm-34kDa peptide were measured in individuals, before and after vector control implementation (VCI). …In the present study, a significant decrease of human IgG response to Nterm-34kDa peptide was observed just one week after VCI.”

5、Establishment of a sandwich light-initiated chemiluminescence assay with double antigen for detecting human cytomegalovirus IgG antibody.
Liu, Z., et al. J Med Virol. 2021 Aug;93(8):5025-5032. doi: 10.1002/jmv.26905. PMID: 33634873.
“Determination of human cytomegalovirus IgG (HCMV IgG) level is of great importance in the diagnosis of HCMV infections. …In this study, a novel, double antigen sandwich homogeneous immunoassay-based light-initiated chemiluminescent assay (LICA) for measuring HCMV IgG serum levels was developed. …This sandwich LICA for HCMV IgG was performed by incubating serum samples with HCMV pp150 protein coated with chemibeads, streptavidin-coated sensibeads, and biotinylated HCMV pp150 protein. …The working conditions of this assay were optimized and the correlation between the results of the LICA and enzyme-linked immunosorbent assay was evaluated. …As a homogeneous immunoassay, this sandwich LICA could accurately and rapidly determine the serum levels of HCMV IgG with a high-throughput. Thus, this newly developed assay could be a useful analytical tool in the clinical diagnosis of HCMV infections.”

For more references about Recombinant Human IgG Antibody Mixes please contact our scientific support team with message@sydlabs.com.

Syd Labs provides the following anti-human Fc blocking reagents:

Anti-human CD16 monoclonal antibody (Clone: 3G8)
Anti-human CD32 monoclonal antibody (Clone: IV.3)
Anti-human CD64 monoclonal antibody (Clone: 22)
Purified human IgG
Recombinant human IgG antibody mixes
Recombinant human IgG isotype control antibodies
Recombinant mouse IgG antibody mixes
Recombinant mouse IgG isotype control antibodies
Recombinant rat IgG antibody mixes
Recombinant rat IgG isotype control antibodies

Syd Labs provides the following anti-mouse Fc blocking reagents:

Anti-mouse CD16/CD32 monoclonal antibody (Clone: 2.4G2)
Recombinant mouse IgG antibody mixes
Recombinant mouse IgG isotype control antibodies
Recombinant rat IgG antibody mixes
Recombinant rat IgG isotype control antibodies

Human IgG Antibody Mix (HyHEL-10) from : Recombinant Human IgG Antibody Mix with more hIgG1: PA007409 Syd Labs

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