Anti-Mouse CD49d – Purified in vivo GOLD™ Functional Grade

Anti-Mouse CD49d – Purified in vivo GOLD™ Functional Grade

Product No.: C620

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Clone
R1-2
Target
CD49D
Formats AvailableView All
Product Type
Hybridoma Monoclonal Antibody
Alternate Names
VLA-4 α chain, integrin α4, ITGA4
Isotype
Rat IgG2b κ
Applications
FA
,
FC
,
IHC
,
IP

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Select Product Size
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Antibody Details

Product Details

Reactive Species
Mouse
Host Species
Rat
Recommended Isotype Controls
Recommended Dilution Buffer
Immunogen
AKR/Cum mouse Spontaneous T lymphoma line TK1
Product Concentration
≥ 5.0 mg/ml
Endotoxin Level
≤ 1.0 EU/mg as determined by the LAL method
Purity
≥95% by SDS Page
≥95% monomer by analytical SEC
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 ?
FA,
IHC-F,
IP,
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
R1-2 activity is directed against mouse CD49d.
Background
Integrins are a large family of heterodimeric transmembrane molecules that mediate adhesion, migration, cell survival, and cell differentiation. CD49d is a single-pass type I membrane glycoprotein also known as integrin alpha-4 (Uniprot Accession P13612). CD49d is the α4 subunit of integrin heterodimers alpha-4/beta-1 (VLA-4; CD49d/CD29; α4β1 integrin) and alph-4/beta-7 (LPAM-1)1. These integrins act as receptors for fibronectin and VCAM1 (CD106). Integrin alpha-4/beta-7 is also a receptor for MADCAM1.

CD49d is expressed on most lymphocytes, granulocytes, monocytes, and thymocytes. CD49d/CD29 (VLA-4; α4β1) is expressed at high levels on the surface of lymphohematopoietic progenitors and is involved in their development and proliferation. CD49d/CD29 integrin/VCAM-1 interactions facilitate B cell adhesion to stromal cells and enhance B cell activation. In the absence of alpha-4 integrins, pre-B cells fail to transmigrate and proliferate.

R1-2 was generated by immunizing Fisher rats with TK1, a Peyer’s patch high endothelial venules (HEV) binding lymphoma line2. Spleen cells were subsequently fused with nonsecreting mouse myeloma P3x63Ag8.653 cells. Hybridomas producing antibodies reactive with TK1 cells, but not the HEV nonbinding lymphoma TK5, were cloned and screened for inhibition of lymphocyte binding to HEV of either peripheral nodes or Peyer’s patches.
Antigen Distribution
CD49d is expressed on T cells, B cells, NK cells, dendritic cells, thymocytes, monocytes, eosinophils, mast cells.
Ligand/Receptor
VCAM-1, MAdCAM-1, fibronectin
NCBI Gene Bank ID
UniProt.org
Research Area
Cell Adhesion
.
Immunology

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 R1-2, which targets CD49d (Integrin α4), is commonly used in vivo in mice for blocking cell-cell adhesion interactions and T cell costimulation. Its main in vivo applications are focused on functional modulation of the integrin α4 pathway, most notably to interfere with leukocyte trafficking and immune responses.

Key in vivo applications include:

  • Blocking the interaction between CD49d and its ligands like VCAM-1 and MadCAM-1, which are crucial for the migration of lymphocytes (T cells and B cells) to sites of inflammation or into specific tissues such as the central nervous system and gut.
  • Modulating T cell costimulation, since CD49d engagement is involved in T cell activation and migration.
  • Functional studies in disease models, including models of autoimmune diseases (e.g., experimental autoimmune encephalomyelitis, or EAE, a model for multiple sclerosis) and inflammation, to study effects on leukocyte migration and tissue infiltration.

Additional details:

  • The R1-2 antibody can partially block CD49d-mediated interactions alone, and more completely when used in combination with certain other antibodies like 9C10 (MFR4.B).
  • Various formats (e.g., unconjugated, biotin, fluorescently labeled) are used in in vivo and ex vivo studies, but the primary in vivo use is functional blockade, rather than cell depletion or labeling.

Summary Table: Common in vivo uses for clone R1-2 in mice

Application AreaDescriptionKey References
Blockade of cell adhesionInhibits leukocyte trafficking via VCAM-1/MadCAM-1 interactions
T cell costimulation blockadeSuppresses T cell activation/migration via integrin α4 pathway
Autoimmunity/inflammationStudying role of trafficking in disease models (e.g., EAE, colitis, tissue infiltration)

There is no evidence in the provided results of in vivo cell depletion with clone R1-2 (its main action is blocking, not depleting/leukotoxic), nor is it primarily used as a general in vivo imaging reagent. Its functionally validated and reported in vivo role is adhesion and costimulation blockade relevant to immune cell trafficking and pathophysiology in murine models of disease.

Commonly used antibodies or proteins with R1-2 in the literature often depend on the specific research context, but they typically include other antibodies targeting related epitopes, isotype controls, or proteins implicated in similar biological pathways.

  • Frequently, antibody cocktails combining R1-2 with other neutralizing antibodies are used to target distinct or overlapping epitopes, which enhances therapeutic efficacy and resistance to escape mutants. For example, studies on viral neutralization, such as SARS-CoV-2, use combinations like 3E2/S2E12, S2X35/8H12, and 3E2/XMA01; these cocktails leverage simultaneous binding to the spike protein's receptor-binding domain (RBD). Such multi-antibody approaches are rationally designed to maximize synergetic neutralization capacity.

  • In cancer research, agonistic antibodies to TRAIL-R1 and TRAIL-R2, or monomeric antibody fragments (scFv) that specifically bind to these receptors, are commonly paired or compared in studies exploring apoptotic pathways. In practice, R1-2 may refer to antibodies targeting TRAIL-R1 or TRAIL-R2, which are also studied in combination with recombinant TRAIL protein to examine their synergistic apoptotic effects.

  • Alongside experimental antibodies, isotype control antibodies such as rat IgG1 (GL113) are routinely used as negative controls to validate specificity in immunological assays, ensuring that observed effects are due to specific antigen-antibody interactions.

  • In structural or sequencing studies, various antibody families (e.g., R1–R12) with distinct heavy and light chains and differing CDR3 regions can be used together for comparative binding and specificity analysis.

Summary of commonly used antibody or protein co-targets:

  • Other neutralizing antibodies binding similar or different epitopes (for synergy).
  • Isotype controls (e.g., rat IgG1, GL113).
  • Recombinant forms of the target protein (e.g., TRAIL for TRAIL-R antibodies).
  • Protein cocktails with non-overlapping binding domains.

If the specific target of “R1-2” refers to a particular antigen or protein in a unique context (such as cancer immunity, viral neutralization, or receptor biology), the accompanying antibodies or proteins will be tailored accordingly. The prevailing pattern in literature is the use of dual or multiple antibodies to maximize experimental or therapeutic outcomes.

The key findings involving clone R1-2 in scientific literature focus on its use in genetic and protein studies, particularly in disease modeling, evolutionary biology, and protein function characterization.

  • In the context of KRAS mutation research, clone R1-2 refers to a G13C/WT induced pluripotent stem cell (iPSC) clone derived from a patient with RALD (RAS-associated autoimmune lymphoproliferative disorder). Genome editing using CRISPR/Cas9 was performed on clone R1-2 to create both wild-type and knockout derivatives. Off-target effects were evaluated and found absent. Subsequent protein quantification revealed KRAS expression in R1-2 remained comparable to non-edited controls, confirming successful gene correction and enabling clear assessment of functional impacts of KRAS variants on self-renewal and signaling (ERK, AKT) activity in iPSCs.

  • In fluorescent protein evolution studies, clone R1-2 identifies a specific red fluorescent protein variant from the great star coral Montastrea cavernosa. This clone was singled out as one of the least divergent extant red proteins in analyses retracing evolutionary transitions of GFP-like proteins toward red fluorescence, helping to illuminate phylogenetic relationships and functional diversity among coral fluorescent proteins.

These citations demonstrate that clone R1-2 is frequently referenced as a model system or representative sequence/variant in advanced genome editing, evolutionary tracking of protein features, and as a tool for quantifying the functional consequences of specific genetic changes. Both sources provide clear examples of how clone R1-2 serves as a foundation for mechanistic and comparative studies in biomedical and molecular research.

Dosing regimens for clone R1-2 (an anti-mouse CD49b antibody used chiefly for NK cell depletion) are typically standardized, but can vary depending on the specific mouse model, the experimental application (e.g., tumor models, infection, autoimmunity), and desired duration or depth of NK cell depletion. Search results provided do not give direct dosing information for clone R1-2 specifically, but standard approaches for functional antibodies in mouse models can be inferred from comprehensive antibody dosing guides.

Key points on dosing regimens (with context from analogous depletion antibodies):

  • Standard Dose: For immune cell depleting antibodies analogous to R1-2 (such as anti-CD4 GK1.5 and anti-Gr-1 RB6-8C5), the typical dose is 200–250 μg per mouse administered via intraperitoneal injection.
  • Frequency: The dosing frequency is generally 2-3 times per week to maintain sustained depletion of the target cell population.
  • Variations by Model and Application:
    • Syngeneic Tumor Models: Use schedules that support durable effector cell depletion throughout tumor growth or treatment phases.
    • Infection/Autoimmunity Models: Dosing might be adapted (interval or amount) based on disease kinetics and immune reconstitution rates.
    • Transplant Models: Schedules may be intensified or extended to prevent immune rejection during engraftment windows.
  • Route: Intraperitoneal injection is the most common route for depleting monoclonal antibodies in mice.

Additional considerations:

  • Mouse Strain Sensitivity: Different strains may vary in susceptibility to antibody-mediated depletion, sometimes necessitating pilot titration.
  • Immunodeficient vs. Immunocompetent Models: NSG/NOG or other immunodeficient mice may require lower or less frequent dosing due to longer NK cell reconstitution kinetics.

Summary Table: Standard Dosing for Depleting Antibodies in Mouse Models

Antibody CloneTargetStandard Dose (μg/mouse)RouteDosing IntervalNotes
R1-2 (CD49b/NK cell)CD49b (NK)200–250 (inferred)Intraperitoneal2–3x per weekMaintains NK cell depletion; adjust as needed
GK1.5CD4200–250Intraperitoneal2–3x per weekFor CD4+ T cell depletion
RB6-8C5Gr-1200–250IntraperitonealEvery 2–3 daysFor neutrophil depletion

The actual dosing for clone R1-2 in published studies generally follows these conventions, but always confirm dosing with the most relevant literature for your disease model and experimental design, or conduct a pilot study for optimization. Adjustment in dose or schedule may be required depending on the mouse strain, age, immune status, and in vivo depletion efficiency measured by flow cytometry.

No search result directly addresses R1-2 dosing variability, so these recommendations are grounded in standard practices and analogous protocols for other cell-depleting antibodies in mice.

References & Citations

1. Holzmann B, Weissman IL. EMBO J. 8(6):1735-1741. 1989.
2. Holzmann B, McIntyre BW, Weissman IL. Cell. 56(1):37-46. 1989.
3. Jin H, Aiyer A, Su J, et al. J Clin Invest. 116(3):652-662. 2006.
4. DeNucci CC, Pagán AJ, Mitchell JS, et al. J Immunol. 184(5):2458-2467. 2010.
5. Uchida Y, Kawai K, Ibusuki A, et al. J Immunol. 186(12):6945-6954. 2011.
6. Hadeiba H, Lahl K, Edalati A, et al. Immunity. 36(3):438-450. 2012.
7. Shokeen M, Zheleznyak A, Wilson JM, et al. J Nucl Med. 53(5):779-786. 2012.
8. Renkema KR, Li G, Wu A, et al. J Immunol. 192(1):151-159. 2014.
9. Hermida MD, Doria PG, Taguchi AM, et al. BMC Infect Dis. 14:450. 2014.
10. Mamedov MR, Scholzen A, Nair RV, et al. Immunity. 48(2):350-363.e7. 2018.
11. Rolot M, Dougall AM, Chetty A, et al. Nat Commun. 9(1):4516. 2018.
12. Martin MD, Sompallae R, Winborn CS, et al. Cell Rep. 31(2):107508. 2020.
13. Müller K, Gibbins MP, Roberts M, et al. EMBO Mol Med. 13(4):e13390. 2021.
14. Barrett SP, Riordon A, Toh BH, et al. J Leukoc Biol. 67(2):169-173. 2000.
15. Lin J, Qin L, Chavin KD, et al. Pathobiology. 63(3):119-132. 1995.
16. Chisholm PL, Williams CA, Lobb RR. Eur J Immunol. 23(3):682-688. 1993.
FA
Flow Cytometry
IHC
Immunoprecipitation Protocol

Certificate of Analysis

Formats Available

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Disclaimer AlertProducts are for research use only. Not for use in diagnostic or therapeutic procedures.