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IgG3

IgG3 (Immunoglobulin G3) is a subclass of the IgG antibody family, which plays a crucial role in the human immune response.
This antibody isotype is characterized by its distinct structural and functional properties, including enhanced effector functions and the ability to activate the complement system.
IgG3 is involved in the neutralization of pathogens, opsonization, and the facilitation of antibody-dependent cell-mediated cytotoxicity.
Researcherss studying IgG3 can utilize the PubCompare.ai platform to optimize their research protocols, identify the best methods for enhanced reproducibility and accuracy, and experience the future of scientific discovery.

Most cited protocols related to «IgG3»

Immunofluorescence detection of CPAF in chlamydia-infected cells was carried out as described previously 67. In brief, HeLa cell monolayer was infected with Chlamydia trachomatis L2 for 30 h. The monolayer, after it was fixed with paraformaldehyde (Sigma-Aldrich) and permeabilized with Saponin (Sigma-Aldrich), was costained with Hoechst 32258 (blue), anti-MOMP antibody MC22 (probed with an FITC-conjugated, mouse IgG3-specific secondary antibody), and anti-CPAFn antibody 54b (probed with a Cy3-conjugated, mouse IgG1-specific secondary antibody). Images were acquired individually for each stain in gray using a Cooker digital camera connected to an AX70 Olympus microscope, and the single-color images were merged in frame into the triple-color image using the software Image Pro.
Publication 2001
Antibodies, Anti-Idiotypic Cells Chlamydia Chlamydia trachomatis Chlorpropamide-Alcohol Flushing Fingers Fluorescein-5-isothiocyanate Fluorescent Antibody Technique HeLa Cells hoechst 32258 IgG1 IgG3 Immunoglobulins Microscopy Mus paraform Reading Frames Saponin Stains
Microtitre plates (Immulon 4HBX, Thermo) were coated with recombinant MSP-119.GST (to which antibodies are predominantly of the IgG1 subclass [36 (link)]) or MSP-2.GST (to which antibodies are predominantly IgG3 [36 (link)]) and blocked with 1% (w/v) skimmed milk powder. Samples were assayed as described previously[36 (link),37 (link)] except that coating antigens, test samples and secondary antibody conjugate were each added in a total volume of 50 μl per well. Ten microliters of the antibody-containing eluate of each spot were added to individual wells of the coated and blocked microtitre plate together with 40 μl blocking buffer to give a final concentration of 1:1,000 with respect to the corresponding plasma sample. Each plate included a five-fold dilution series (1:50 to 1:156,250 final dilutions) of a standard African hyperimmune plasma pool. Bound antibodies were detected with either rabbit anti-human-IgG -HRP (Dako, Ely, UK), or sheep-anti-human IgG1 or IgG3-HRPconjugates (The Binding Site, Birmigham, UK) secondary antibodies and developed with o-phenylenediamine-H2O2.
A titration curve was fitted to the ODs obtained for the standard plasma dilutions by least squares minimisation using a three variable sigmoid model and the solver add-in in Excel (Microsoft), assuming an arbitrary value of 1000 Units/ml of antibody against each antigen in the standard pool. OD values for the spot extracts were converted to units/ml using this fitted curve.
Recoveries for blood spots were estimated as follows (full details in Additional file 1): serum or plasma ODs were converted to concentrations as above, the concentrations were multiplied by a recovery factor and then converted back to 'corrected' ODs – the ODs which would have been obtained if the serum or plasma had been more dilute. The value of the recovery factor was then optimized by weighted least squares minimisation comparing the actual ODs for the blood spots and the corrected OD values for serum or plasma, using the solver add-in in Excel™.
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Publication 2008
1,2-diaminobenzene anti-IgG Antibodies Antigens Binding Sites BLOOD Buffers Exanthema factor A Homo sapiens IgG1 IgG3 Immunoglobulins Milk, Cow's Negroid Races Peroxide, Hydrogen Plasma Powder Rabbits Serum Sheep Sigmoid Colon Technique, Dilution Titrimetry
Genomic DNA was isolated from B cells, either resting or cultured with LPS for 4 d. Cell pellets were incubated with proteinase K (0.5 mg/ml), RNaseA (100 μg/ml), and SDS (0.5%) in STE (0.1 M NaCl, 20 mM Tris, 1 mM EDTA) for 2 h at 37°C, followed by 3–4 extractions with phenol/chloroform (1:1) and precipitation with 0.3 M sodium acetate, pH 7, and ethanol. DNA was wound out on glass rods and resuspended in TE, pH 8. The germline Sγ3 segment was amplified by PCR from resting purified B cells from WT(129 × B6) mice for comparison to Sμ-Sγ3 junctions from cells induced to switch to IgG3. Expand HiFidelity Taq polymerase (Roche Laboratories) was used with the following primers: g3–1 (5′CAGGCTAAGATGGATG- CTACAGGG-3′) (MUSIGHANA 404–427) and g3–2 (5′TAC- CCTGACCCAGGAGCTGCATAAC-3′) (MUSIGHANA 2603–2628) to amplify the 2.22-kb fragment of germline Sγ3. Sμ-Sγ3 junctions were amplified by PCR using Expand Long Template Taq polymerase (Roche Laboratories) and the primers μ3-H3 (5′AACAAGCTTGGCTTAACCGAGATGAGCC-3′) and g3–2 (above). The germline Sμ sequence was deduced by comparing the sequences of a large number of Sμ-Sγ3 junctions from wild-type mice. For the sequence analyses, the wild-type sequences from the corresponding littermates were used.
Publication 2002
B-Lymphocytes Cells Chloroform Edetic Acid Endopeptidase K Ethanol Genome Germ Line IgG3 Mice, 129 Strain Mus Oligonucleotide Primers Pellets, Drug Phenols Rod Photoreceptors Sequence Analysis Sodium Acetate Sodium Chloride Taq Polymerase Tromethamine Wounds
Antibodies used for flow cytometry were from eBioscience (San Diego, CA) unless otherwise indicated. Cell suspensions were incubated with FITC-labeled anti-GL7 (BD Biosciences, San Jose, CA), PercP-Cy5.5-labeled anti-B220, APC-labeled anti-mouse IgM, Alexa Fluor 700-labeled anti-CD38, and APC-eFluor780-labeled anti-CD4, CD8, F480, CD11c, and Gr-1 antibodies. The cells were then fixed in 2% formaldehyde for 20 minutes in ice, permeabilized with 0.5% saponin (Sigma), and incubated with biotin-labeled anti-IgG1 (A85-1), IgG2c (5.7), IgG2b (R12-3), IgG3(R40-82) (all from BD Biosciences), IgA (RMA-1), and IgE (RME-1) antibodies (both from Biolegend, San Diego, CA) followed by Pacific Blue-labeled streptavidin and Pacific Orange labeled anti-mouse Ig H+L antibody (both from Invitrogen). In some cases, FITC-labeled anti-IgD, CD80, CD73, VLA-4, CD16/32 (BD Biosciences), or CD44 antibodies were used instead of anti-GL7 antibody. In other cases, FITC-labeled anti-B220, PercP-Cy5.5-labeled anti-IgM, and APC-labeled anti-TACI or anti-BAFFR antibodies were substituted for FITC-labeled anti-GL7, PercP-Cy5.5-labeled anti-B220, and APC-labeled anti-mouse IgM antibodies. For detection of BrdU, samples were subjected to an additional fixation with Cytofix/Cytoperm (BD Biosciences), followed by incubation with 1 mg/ml DNAse I (Sigma) for 60 minutes at 37°C, culture supernatant containing 24G2 antibody plus 1% rat serum for 10 minutes at 4°C, and FITC-labeled anti-BrdU (BD Biosciences) for 30 minutes at 4°C. For marginal zone and B1 cell detection, cell suspensions were prepared in the absence of collagenase to prevent cleavage of CD23 (5 (link)), and incubated with FITC-labeled anti-CD23 antibody (BD), PercP-Cy5.5-labeled anti-B220, APC-labeled anti-CD93, Alexa Fluor 650-labeled anti-IgM, eFluor-450-labeled anti-CD21, biotin-labeled CD43 (BD), and APC-eFluor780-labeled anti-CD4, CD8, F480, CD11c, and Gr-1 antibodies, followed by an incubation with V500-labled streptavidin (BD). The cells were then fixed in 2% formaldehyde, permeabilized with 0.5% saponin (Sigma), and incubated with Alexa Fluor 350-labeled anti-mouse Ig H+L antibody. Flow cytometry was performed on a 4-laser (355nm, 405nm, 488nm, 633nm) LSR II device (BD Biosciences) and analyzed with FlowJo software (Tree Star, Ashland, OR). Fluorescent AccuCheck counting beads (Invitrogen) were used to calculate total numbers of live lymphocytes in the column bound and flow through suspensions.
Publication 2011
Alexa 350 Anti-Antibodies anti-IgD anti-IgM Antibodies Antibodies, Anti-Idiotypic Be1 Cells Biotin Bromodeoxyuridine CD44 protein, human Cells Collagenase CY5.5 cyanine dye Cytokinesis Deoxyribonucleases Flow Cytometry Fluorescein-5-isothiocyanate Formaldehyde IgG1 IgG2B IgG3 Integrin alpha4beta1 lumiliximab Lymphocyte Count Medical Devices Mus NT5E protein, human Saponin Serum SPN protein, human Streptavidin TNFRSF13B protein, human Trees
Fig. S1 depicts analyses of lymphocyte development in Vav2ko and Vav3ko mice. Flow cytometry analyses of thymocytes, splenocytes, LN, and BM cells from WT mice (WT), Vav2ko (Vav2), or Vav3ko (Vav3) mice are shown. Fig. S2 shows the strategy for generation of Vav3-deficient mice. Southern blotting analyses of targeted ES cells used to generate chimeric mice and their offspring and Western blotting analyses of Vav3ko splenocyte lysates with Vav3-specific antibodies showing no full-length or truncated Vav3 protein present are also shown. Fig. S3 presents analyses of humoral TD responses in Vav3-deficient mice. Concentrations of TNP-specific IgM, IgG1, IgG2b, and IgG3 antibodies analyzed at various time points post-primary or secondary immunization with TNP-conjugated keyhole hemagglutinin protein (TNP-KLH) are shown. Fig. S4 presents analyses of cytokine production by Vav-deficient T cells showing defective IL-2 and IFNγ production. Figs. S1–4 are available online at http://www.jem.org/cgi/content/full/jem.20030874/DC1.
Publication 2003
Antibodies Cells Chimera Cytokine Figs Flow Cytometry Hemagglutinin IgG1 IgG2B IgG3 Interferon Type II Lymphocyte Mus Proteins Secondary Immunization Southern Blotting T-Lymphocyte Thymocyte trinitrophenyl keyhole limpet hemocyanin Western Blot

Most recents protocols related to «IgG3»

First, the 96-well plates were coated with 100 µL of B. abortus A19 LPS (100 μg/mL) and incubated overnight at 4°C. Then, the plates were washed with PBST 3 times, and 200 μL of blocking buffer (5% skim milk in PBST) was added to each well and incubated at 37°C for 2 h. After blocking, sera from each mouse were added to the corresponding wells and serially diluted with dilution buffer (10% blocking buffer) and incubated at 37°C for 1 h. The plates were washed and 100 μL of HRP-conjugated goat anti-mouse IgG, IgG1, IgG2a, IgG2b, or IgG3 antibody (Abcam, Cambridge, MA, United States) (1:15,000) was added to each well and incubated at 37°C for 1 h. After washing, the Soluble TMB Kit (CWBio, Beijing, China) was used for color development, and the absorption at a wavelength of 450 nm was measured using a microplate spectrophotometer.
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Publication 2023
anti-IgG Buffers Fetuses, Aborted Goat IgG1 IgG2A IgG2B IgG3 Immunoglobulins Mice, Laboratory Milk, Cow's Serum Technique, Dilution
Complement proteins FH (#341274), FB (#341262), C3b (#204860), C3 (#204885), factor I (FI) (#341280), C1q (#204876), goat anti-FB (#341272) and goat anti-FH (#341276) antiserum were obtained from Merck (Darmstadt, Germany; obtained via Merck Life Science Kft., Budapest, Hungary). FD (#A409), properdin (#A412), mouse anti-C5b-9 (#A239), anti-FB (Bb) (#A227), anti-FB (Ba) (#A225) and C3a EIA kit (#A032) were from Quidel (obtained via Biomedica, Budapest, Hungary). Purified human IgG (#I2511), human serum albumin (#A3782), human alpha-1 antitrypsin (#SRP6312), antibodies against C1q (#234390), human IgG (#A6029), IgM (#A0420), IgA (#A0295), IgG1 (clone HP-6001, #I9388), IgG2 (clone HP-6002, #I9513), IgG3 (clone HP-6050, #I7260), IgG4 (clone HP-6025, #I7385), IgGκ (clone KP-53, #K4377), IgGλ (clone HP-6054, #L6522), and avidin-HRP (#A7419) were purchased from Merck. Mouse IgG1 (clone MOPC-21, #BZ-400101) was obtained from BioLegend (San Diego, CA). MBL (#9086-MB) and biotin-conjugated goat anti-MBL (#BAF2307) were from R&D Systems (Minneapolis, MN), HRP-conjugated goat anti-mouse (#P0447), rabbit anti-goat (#P0449) and swine anti-rabbit (#P0217) antibodies were from DAKO (Hamburg, Germany). Goat anti-C3 F(ab’)2 (#55062) and HRP-conjugated goat anti-C3 F(ab’)2 (#55237) were from MP Biomedicals (Solon, OH).
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Publication 2023
Antibodies avidin-horseradish peroxidase complex Biotin Clone Cells Complement Membrane Attack Complex Complement System Proteins Fibrinogen Goat Homo sapiens IgG1 IgG2 IgG3 IgG4 Immune Sera Mus Properdin Rabbits SERPINA1 protein, human Serum Albumin, Human Solon Sus scrofa
Autoantibodies were detected using ELISA method, as described by Józsi and Uzonyi (36 (link)). Briefly, FH, FB, C3b, FI in 5 µg/ml, MBL and C1q in 2 µg/ml and HSA or alpha-1 antitrypsin as negative controls were immobilized on microtiter plates. After blocking with 5% BSA in DPBS-0.1% Tween-20, serum samples were added in DPBS-0.1% Tween-20 at a dilution of 1:50 for 1 hour at RT. To detect autoantibodies against solid phase C3 convertase of the alternative pathway (C3bBbP), the convertase was built up (see below) and incubated with serum samples diluted 1:50. The bound autoantibodies were detected with HRP-conjugated goat anti-human IgG, anti-human IgA or anti-human IgM. Color reaction was developed by adding TMB solution (BioLegend). After stopping with H2SO4, absorbance was measured at 450 nm and at 620 nm as reference wavelength. To detect anti-MBL autoantibodies, serum samples were diluted in DPBS containing 10 mM EDTA. In the case of detecting anti-C1q autoantibodies, serum samples were diluted in DPBS containing 1 M NaCl to avoid interaction of C1q globular heads with IgG Fc parts. Samples were accepted as positive if the mean OD value for the investigated protein was at least two-fold greater than the mean OD value for the negative control protein alpha-1 antitrypsin. The isotypes of the autoantibodies were determined using monoclonal antibodies specific for IgG1, IgG2, IgG3, IgG4, IgGκ and IgGλ.
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Publication 2023
anti-IgA anti-IgG anti-IgM Autoantibodies Complement C3 Convertase, Alternative Pathway Edetic Acid Enzyme-Linked Immunosorbent Assay Goat Head Homo sapiens IgG1 IgG2 IgG3 IgG4 Immunoglobulin Isotypes Monoclonal Antibodies Proteins SERPINA1 protein, human Serum Sodium Chloride Technique, Dilution Tween 20
For isolation of IPF and control MPCs, primary IPF mesenchymal cells were labeled with mouse anti–human SSEA4 antibody conjugated to Alexa Fluor 647 (clone MC-813-70; catalog 560796; BD Biosciences) and mouse anti–human CD44 conjugated to FITC (clone IM7; catalog 103006; BioLegend). Cells were sorted on a FACS Aria Cell Sorter (BD Biosciences). Cells that were SSEA4+ and CD44+ (relative to mouse IgG3 κ isotype control conjugated to Alexa Fluor 647 and mouse IgM κ isotype control conjugated to FITC, respectively) (clone J606, catalog 560803 [BD Biosciences] and catalog 402207 [BioLegend]) were collected. To generate sufficient numbers of MPCs for the in vivo mouse studies, SSEA4+CD44+ cells were expanded by culture in DMEM + 10% FCS for 7 days prior to use. The resulting MPC cultures were reanalyzed for SSEA4 expression by FACS analysis and for colony formation in vitro. In total, 97% of day 7 MPCs were SSEA4+ and formed colonies in methylcellulose indicating retention of progenitor self-renewal properties during in vitro expansion.
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Publication 2023
Alexa Fluor 647 CD44 protein, human Cells Clone Cells Fluorescein-5-isothiocyanate Homo sapiens IgG3 Immunoglobulin Isotypes Immunoglobulins isolation Mesenchymal Stem Cells Methylcellulose Mus NRG1 protein, human Retention (Psychology) stage-specific embryonic antigen-4
Standardized ELISAs were used to quantify serum RH5.1–specific IgG1, IgG2, IgG3, IgG4, IgA, IgA1, IgA2, and IgM responses in vaccinees. Nunc MaxiSorp flat-bottom ELISA plates (44-2404-21, Invitrogen) were coated overnight with 5 μg/mL of RH5.1 protein in PBS. Plates were washed with washing buffer composed of PBS containing 0.05% TWEEN 20 (P1379, Sigma-Aldrich) and blocked with 100 μL of Blocker Casein in PBS (37582, Thermo Fisher Scientific). After removing blocking buffer, standard curve and internal controls were created in casein using a pool of high-titre volunteer plasma, specific for each isotype or subclass being tested, and 50 μL of each dilution was added to the plate in duplicate. Test samples were diluted in casein to a minimum dilution of 1:50, and 50 μL was added in triplicate. Plates were incubated for 2 hours at 37°C and washed in washing buffer. An alkaline phosphatase–conjugated secondary antibody from Southern Biotech was diluted at the manufacturer’s recommended minimum dilution for ELISA in casein. The antibody used was dependent on the isotype or subclass being assayed and were as follows: mouse anti–human IgG1 Fc-AP (catalog 9054-04), IgG2 Fc-AP (catalog 9060-04), IgG3 hinge-AP (catalog 9210-04), IgG4 Fc-AP (catalog 9200-04), IgA1-AP (catalog 9130-04), and IgA2-AP (catalog 9140-04) and goat anti–human IgA-AP (catalog 2050-04) and IgM-AP (catalog 2020-04). In total, 50 μL of the secondary antibody dilution was added to each well of the plate and incubated for 1 hour at 37°C. Plates were developed using PNPP alkaline phosphatase substrate (N2765, Sigma-Aldrich) for 1–4 hours at 37°C. Optical density at 405 nm was measured using an ELx808 absorbance reader (BioTek) until the internal control reached an OD405 of 1. The reciprocal of the internal control dilution giving an OD405 of 1 was used to assign an AU value of the standard. Gen5 ELISA software v3.04 (BioTek) was used to convert the OD405 of test samples into AU values by interpolating from the linear range of the standard curve fitted to a 4-parameter logistics model. Any samples with an OD405 below the linear range of the standard curve at the minimum dilution tested were assigned a minimum AU value according to the lower limit of quantification of the assay.
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Publication 2023
4-aminophenylphosphate Alkaline Phosphatase anti-IgA ATP8A2 protein, human Biological Assay Buffers Caseins Enzyme-Linked Immunosorbent Assay Goat Homo sapiens IgA1 IgA2 IgG1 IgG2 IgG3 IgG4 Immunoglobulin Isotypes Immunoglobulins Mus Plasma Serum Technique, Dilution Tween 20 Vision Voluntary Workers

Top products related to «IgG3»

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IgG2b is a type of immunoglobulin G (IgG) subclass that is commonly used in laboratory research and applications. It functions as an antibody, playing a role in the immune system's response to foreign substances. IgG2b has specific structural and functional characteristics that make it a valuable tool for various experimental and analytical purposes.
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IgG2c is a type of immunoglobulin G (IgG) antibody. It is a laboratory product that can be used for various research and analytical applications involving the detection and quantification of IgG2c in biological samples.
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IgG2a is an immunoglobulin subclass that functions as an antibody. It is produced by plasma B cells and plays a role in the adaptive immune response.
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Bovine serum albumin (BSA) is a common laboratory reagent derived from bovine blood plasma. It is a protein that serves as a stabilizer and blocking agent in various biochemical and immunological applications. BSA is widely used to maintain the activity and solubility of enzymes, proteins, and other biomolecules in experimental settings.
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IgG2a is a type of immunoglobulin G (IgG) antibody. It is a component of the adaptive immune system and plays a role in the humoral immune response. IgG2a is involved in the neutralization of toxins and viruses, as well as the activation of the complement system.
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The FACSCalibur is a flow cytometry system designed for multi-parameter analysis of cells and other particles. It features a blue (488 nm) and a red (635 nm) laser for excitation of fluorescent dyes. The instrument is capable of detecting forward scatter, side scatter, and up to four fluorescent parameters simultaneously.
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IgG2b is an immunoglobulin subclass that plays a role in the immune system. It is a component of the antibody response. IgG2b has core functions related to immune system processes, but a detailed description cannot be provided while maintaining an unbiased and factual approach without extrapolation.
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The LPS laboratory equipment is a high-precision device used for various applications in scientific research and laboratory settings. It is designed to accurately measure and monitor specific parameters essential for various experimental procedures. The core function of the LPS is to provide reliable and consistent data collection, ensuring the integrity of research results. No further details or interpretations can be provided while maintaining an unbiased and factual approach.
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The FACSCanto II is a flow cytometer instrument designed for multi-parameter analysis of single cells. It features a solid-state diode laser and up to four fluorescence detectors for simultaneous measurement of multiple cellular parameters.
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ELISA (Enzyme-Linked Immunosorbent Assay) plates are a type of laboratory equipment used for performing enzyme-linked immunosorbent assays. These plates typically consist of a flat surface with multiple wells, allowing for the simultaneous analysis of multiple samples or conditions. The wells are designed to facilitate the binding of specific proteins or other molecules to the plate surface, enabling the detection and quantification of target analytes in the samples.

More about "IgG3"

Immunoglobulin G3 (IgG3) is a crucial subtype of the IgG antibody family, playing a vital role in the human immune response.
This antibody isotype is characterized by its distinct structural and functional properties, including enhanced effector functions and the ability to activate the complement system.
IgG3 is involved in the neutralization of pathogens, opsonization, and the facilitation of antibody-dependent cell-mediated cytotoxicity (ADCC).
Researchers studying IgG3 can leverage the power of the PubCompare.ai platform to optimize their research protocols and identify the best methods for enhanced reproducibility and accuracy.
This AI-driven platform compares protocols from literature, pre-prints, and patents, helping researchers locate the ideal IgG3 research procedures with ease and confidence.
In addition to IgG3, researchers may also be interested in exploring other IgG subtypes, such as IgG2b, IgG2c, and IgG2a.
These antibody isotypes exhibit distinct functional properties and can play important roles in various immune responses.
Furthermore, researchers may utilize bovine serum albumin (BSA) as a blocking agent in IgG3 assays, such as flow cytometry using a FACSCalibur or FACSCanto II instrument, or in enzyme-linked immunosorbent assays (ELISAs) on ELISA plates.
Understanding the role of IgG3 and its interactions with other immune components, like lipopolysaccharide (LPS), can provide valuable insights into the complexities of the human immune system.
By optimizing their research protocols and leveraging the insights gained from the PubCompare.ai platform, researchers can advance their understanding of IgG3 and its implications in various disease states and therapeutic interventions.