Anti-Mouse MHC Class II (I-A) [Clone Y-3P] – Purified in vivo GOLD™ Functional Grade

Anti-Mouse MHC Class II (I-A) [Clone Y-3P] – Purified in vivo GOLD™ Functional Grade

Product No.: H470

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Clone
Y-3P
Target
MHC class II (I-A)
Formats AvailableView All
Product Type
Hybridoma Monoclonal Antibody
Isotype
Mouse IgG2a k
Applications
B
,
FC

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Antibody Details

Product Details

Reactive Species
Mouse
Host Species
Mouse
Recommended Dilution Buffer
Immunogen
BALB/c x C57BL/6 F1 mouse spleen cells
Product Concentration
≥ 5.0 mg/ml
Endotoxin Level
< 1.0 EU/mg as determined by the LAL method
Purity
≥95% monomer by analytical SEC
>95% by SDS Page
Formulation
This monoclonal antibody is aseptically packaged and formulated in 0.01 M phosphate buffered saline (150 mM NaCl) PBS pH 7.2 - 7.4 with no carrier protein, potassium, calcium or preservatives added. Due to inherent biochemical properties of antibodies, certain products may be prone to precipitation over time. Precipitation may be removed by aseptic centrifugation and/or filtration.
State of Matter
Liquid
Product Preparation
Functional grade preclinical antibodies are manufactured in an animal free facility using only in vitro protein free cell culture techniques and are purified by a multi-step process including the use of protein A or G to assure extremely low levels of endotoxins, leachable protein A or aggregates.
Storage and Handling
Functional grade preclinical antibodies may be stored sterile as received at 2-8°C for up to one month. For longer term storage, aseptically aliquot in working volumes without diluting and store at ≤ -70°C. Avoid Repeated Freeze Thaw Cycles.
Regulatory Status
Research Use Only
Country of Origin
USA
Shipping
2 – 8° C Wet Ice
Additional Applications Reported In Literature ?
B,
FC
Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change.

Description

Description

Specificity
Y-3P activity is directed against mouse MHC class II (I-A) glycoprotein antigens, including the haplotypes I-Ab, I-Af, I-Ap, I-Aq, I-Ar, I-As, I-Au, I-Av, and weakly I-Ak. Y-3P also reacts with the equivalent complexes in rats.
Background
H-2, the murine major histocompatibility complex (MHC), is composed of a diverse group of antigens divided into class I and II proteins that function in immune response1. Class II molecules, also known as Ia antigens, regulate recognition of foreign antigens on the surfaces of antigen presenting cells and play a major role in the mixed lymphocyte response2. Mice have two class II isotypes, I-A and I-E, each of which is a glycoprotein composed of an ⍺ and β subunit. The N-terminal α1 and β1 domains of the MHC class II isotype form the antigen- binding groove, which binds 13-25 aa peptides derived from exogenous antigens3.

On APCs, MHC class II molecules play a critical role in the adaptive immune response by presenting phagocytosed antigens to helper CD4 T cells. The T cell receptor (TCR)/CD3 complex of CD4 T cells interacts with peptide-MHC class II, which induces CD4 T cell activation leading to the coordination and regulation of other effector cells. CD4 molecules also bind to MHC class II, which helps augment TCR signaling4. Additionally, MHC class II expressed on activated T cells are capable of antigen presentation5 and can transduce signals into T cells, enhancing T cell proliferation and activity6.

Y-3P was generated by repeatedly immunizing primed mice with activated T cells over the course of a year7. Y-3P reacts with I-A subregion-controlled A ⍺: A β complexes of all mouse strains except the responder strain H-2d. Y-3P is commonly used for in vivo blockade of TCR stimulation8,9 and MHC class II blocking10,11,12,13,14,15.

Antigen Distribution
MHC class II molecules are constitutively expressed on professional antigen-presenting cells (APCs), including macrophages, monocytes, dendritic cells (DCs), and B cells, and are induced on T cells upon activation.
Ligand/Receptor
CD3/TCR, CD4
NCBI Gene Bank ID
UniProt.org
Research Area
Immunology
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Innate Immunity

Leinco Antibody Advisor

Powered by AI: AI is experimental and still learning how to provide the best assistance. It may occasionally generate incorrect or incomplete responses. Please do not rely solely on its recommendations when making purchasing decisions or designing experiments.

Clone Y-3P is widely used in mice for in vivo blockade of MHC class II (I-A) molecules, primarily to inhibit I-A-restricted T cell responses or manipulate antigen presentation, making it a key tool in immunological blockading and functional studies.

Common in vivo applications include:

  • Blocking MHC Class II-mediated antigen presentation: Y-3P is administered to mice to prevent antigen-presenting cells from activating CD4+ T cells via I-A MHC class II molecules, thereby inhibiting immune responses dependent on this pathway.
  • Immunological tolerance and autoimmunity research: Used in models such as experimental autoimmune encephalomyelitis (EAE), Y-3P can help dissect mechanisms of autoimmune disease by blocking MHC II-dependent T cell activation.
  • Functional studies of immune response: By blocking TCR stimulation through MHC II, researchers can isolate and study the roles of CD4+ T cells, antigen-presenting cells, and specific antigenic pathways in various models of infection, transplantation, and immune regulation.
  • MHC II immunopeptidomics and molecular studies: Y-3P facilitates the study of peptide presentation and antigen repertoire by blocking or isolating specific I-A haplotypes in vivo and ex vivo.
  • Studying tolerance induction and pathogen clearance: The antibody is used in tolerance models to understand the requirement for MHC II-TCR interaction during tolerance induction or immune clearance of pathogens.

Summary table:

ApplicationPurpose
In vivo blocking of MHC class II (I-A)Inhibit I-A-restricted T cell responses and antigen presentation
Autoimmunity and tolerance models (e.g., EAE)Block T cell activation to study disease/tolerance mechanisms
Functional immune studiesDissect roles of CD4+ T cells and APCs in vivo
MHC II immunopeptidomicsIsolate/analyze antigen peptides presented by I-A
Manipulating immune responsesInduce or prevent immune reactions in transplantation, infection

Y-3P is reported to react with various I-A haplotypes (I-A^b^, I-A^f^, I-A^p^, etc.), allowing use across multiple mouse strains. Its specificity is to the I-A subtype, not I-E. Typical in vivo applications focus on immune blockade experiments where antigen-specific MHC II:T cell interactions need to be interrupted.

These uses are broadly supported by product documentation, research antibody suppliers, and peer-reviewed studies using Y-3P for in vivo immunological modulation.

Commonly, Y-3P (an anti-mouse MHC class II antibody) is used alongside antibodies and proteins targeting T cell activation and differentiation, especially in immunological studies focused on antigen presentation and T cell responses.

Frequently co-used antibodies and proteins include:

  • Anti-CD3: Marks the T cell receptor complex, essential for analyzing or stimulating T cell activation as it engages with the MHC class II/TCR axis.
  • Anti-CD28: Provides the co-stimulatory signal necessary for full T cell activation, often included in studies assessing T cell function or activation.
  • Anti-CD4: Identifies helper T cells and is involved in co-receptor signaling with MHC class II molecules.
  • Markers for antigen-presenting cells (APCs): This includes markers for macrophages, dendritic cells, monocytes, and B cells, as these cells constitutively express MHC class II and are integral to studies using Y-3P.

In experimental setups, researchers often combine Y-3P with these antibodies in:

  • Assessing T helper cell activation
  • Blocking assays to dissect MHC class II–dependent T cell responses
  • Flow cytometry panels evaluating APC or T cell populations.

Other relevant antibodies can include isotype controls (e.g., Mouse IgG2a), specific peptide:MHC II–recognizing antibodies such as Y-Ae (for Eα:IA^b complexes), or anti-MHC II antibodies that recognize different haplotypes for comparative or blocking studies.

These combinations are essential for characterizing cell interactions within the immune response and for functional studies of antigen presentation, tolerance, and transplantation immunology.

Clone Y-3P is a widely used monoclonal antibody that targets mouse MHC class II (I-A) molecules, and its citations in scientific literature reveal several key applications and findings:

  • Blockade of MHC Class II and TCR Stimulation: Y-3P is frequently employed for in vivo blockade of MHC class II, effectively interrupting antigen presentation and T cell receptor (TCR) stimulation in mice. This blockade is crucial for dissecting the roles of MHC class II molecules in immune responses, tolerance, and autoimmunity.

  • Inhibition of T Cell Responses: The antibody is reported to inhibit I-A-restricted T cell responses in functional assays. This allows researchers to specifically assess the MHC class II dependence of immune phenomena by blocking the interaction between antigen-presenting cells and CD4+ T cells.

  • Assessment of MHC II Expression: Y-3P is commonly used as a detection reagent in flow cytometry and immunostaining to assess and quantify the expression of MHC class II molecules (I-A) on the surface of various cell types, including B cells and dendritic cells. This is critical for studies examining immune cell phenotypes and dynamics.

  • Immunological Blockade Experiments: The antibody is a standard tool for in vivo immunological blockade experiments, particularly in mouse models of disease where the function of MHC class II needs to be transiently eliminated to understand its contribution to pathogenesis or tolerance.

  • High Citation in Foundational Immunology Studies: Y-3P is cited in numerous foundational studies investigating T cell selection, antigen processing, autoimmunity, and self-peptide presentation by MHC class II molecules (for example: positive selection in the thymus, CNS immune privilege, and formation of peptide-MHC complexes).

  • Detection of Empty vs. Loaded MHC Molecules: The antibody has also been instrumental in recognizing both loaded and "empty" MHC class II molecules, assisting in research on peptide loading and editing within antigen-presenting cells.

In summary, Y-3P is a foundational tool antibody in mouse immunology, providing specific blockade, detection, and mechanistic interrogation of MHC class II (I-A)-mediated processes throughout the literature. Its versatility and specificity are reflected in many landmark immunological publications.

Dosing regimens for the Y-3P clone, which targets mouse MHC class II (I-A), can vary significantly across different mouse models. The dosing strategy often depends on several factors, including:

  • Experimental Objective: Whether the goal is to deplete certain cells, functionally inactivate MHC class II, or interfere with antigen presentation.
  • Route of Administration: While intraperitoneal (i.p.) injections are commonly used, other routes like intravenous (i.v.) or subcutaneous (s.c.) may also be employed depending on the study design.
  • Mouse Strain: Different strains may have varying responses to the antibody due to genetic differences affecting immune responses.

A typical dosing range for functionally similar MHC class II blocking antibodies, such as Y-3P, is generally between 100–250 μg per mouse. However, specific dosing regimens in the literature may vary based on the experimental design and desired outcomes. Investigators often need to determine their own optimal working dilution and dosing schedule based on the specific requirements of their study.

For precise dosing, it is crucial to consult the specific literature related to the mouse model being used, as dosing can significantly impact the efficacy and relevance of experimental results.

References & Citations

1. Yoshida R. Adv Immunol. 124:207-247. 2014.
2. Spencer JS, Kubo RT. J Exp Med. 169(3):625-640. 1989.
3. Wieczorek M, Abualrous ET, Sticht J, et al. Front Immunol. 8:292. 2017.
4. Artyomov MN, Lis M, Devadas S, et al. Proc Natl Acad Sci USA. 107(39):16916-16921. 2010.
5. Barnaba V, Watts C, de Boer M, et al. Eur J Immunol. 24(1):71-75. 1994.
6. Di Rosa F, D'Oro U, Ruggiero G, et al. Hum Immunol. 38(4):251-260. 1993.
7. Janeway CA Jr, Conrad PJ, Lerner EA, et al. J Immunol. 132(2):662-667. 1984.
8. Feng Y, van der Veeken J, Shugay M, et al. Nature. 528(7580):132-136. 2015.
9. Campisi L, Barbet G, Ding Y, et al. Nat Immunol. 17(9):1084-1092. 2016.
10. Stefanová I, Dorfman JR, Germain RN. Nature. 420(6914):429-434. 2002.
11. Andersson J, Stefanova I, Stephens GL, et al. Int Immunol. 19(4):557-566. 2007.
12. Younes SA, Punkosdy G, Caucheteux S, et al. PLoS Biol. 9(10):e1001171. 2011.
13. Guo L, Huang Y, Chen X, et al. Nat Immunol. 16(10):1051-1059. 2015.
14. Kawabe T, Yi J, Kawajiri A, et al. Nat Commun. 11(1):3366. 2020.
15. Kruse B, Buzzai AC, Shridhar N, et al. Nature. 618(7967):1033-1040. 2023.
16. Wei J, Loke P, Zang X, et al. J Exp Med. 208(8):1683-1694. 2011.
17. Alspach E, Lussier DM, Miceli AP, et al. Nature. 574(7780):696-701. 2019.
18. Hos BJ, Tondini E, Camps MGM, et al. Cell Rep. 41(2):111485. 2022.
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