The RTK2118 antibody (IgM, kappa) is suitable for use as an isotype-matched control antibody in several applications to estimate the degree of non-specific binding by an antigen-specific antibody. Ideally, the isotype control should have the same subclass of heavy chain (IgA, IgD, IgE, IgG, or IgM) and light chain (kappa or lambda) as the specific antibody being used. If a conjugated antibody is used, an isotype control conjugated to the same molecule (e.g. fluorochrome) should be chosen. The use of an appropriate isotype control helps confirm the specificity of the antigen-specific antibody and indicates non-specific binding that may result from binding to Fc receptors or other cell components.
This antibody clone has been verified for use as an isotype control antibody for assessing non-specific binding to cells in applications such as flow cytometry and immunofluorescence microscopy applications (surface and intracellular staining).
This antibody clone has been verified for use as an isotype control antibody for assessing non-specific binding to cells in applications such as flow cytometry and immunofluorescence microscopy applications (surface and intracellular staining).
Data Figures
Figure 1. Data for Unconjugated
(A) Flow cytometry analysis of C57BL/6 mouse splenocytes labeled with Rat IgM, kappa Isotype Control Antibody, Clone RTK2118, followed by a mouse anti-rat IgM antibody, FITC and Anti-Mouse CD45 Antibody, Clone 30-F11, PE (Catalog #60030PE). (B) Flow cytometry analysis of C57BL/6 mouse splenocytes labeled with Anti-Mouse CD49b Antibody, Clone DX5 (Catalog #60020), followed by a mouse anti-rat IgM antibody, FITC and Anti-Mouse CD45 Antibody, Clone 30-F11, PE. (C) Human ES cells were cultured in mTeSR™1 on Corning® Matrigel®-coated glass slides, then fixed and stained with Anti-Mouse SSEA-3 Antibody, Clone MC-631 (Catalog #60061), followed by goat anti-mouse IgG, FITC. Inset shows cells labeled with Rat IgM, kappa Isotype Control Antibody, Clone RTK2118 followed by goat anti-mouse IgG, FITC.
Figure 2. Data for Alexa Fluor® 488-Conjugated
(A) Flow cytometry analysis of C57BL/6 mouse splenocytes labeled with Rat IgM, kappa Isotype Control Antibody, Clone RTK2118, Alexa Fluor® 488 and Anti-Mouse CD45 Antibody, Clone 30-F11, APC (Catalog #60030AZ). (B) Flow cytometry analysis of C57BL/6 mouse splenocytes labeled with Anti-Mouse CD49b Antibody, Clone DX5, Alexa Fluor® 488 (Catalog #60020AD) and Anti-Mouse CD45 Antibody, Clone 30-F11, APC.
Figure 3. Data for APC-Conjugated
(A) Flow cytometry analysis of C57BL/6 mouse splenocytes labeled with Rat IgM, kappa Isotype Control Antibody, Clone RTK2118, APC and Anti-Mouse CD45 Antibody, Clone 30-F11, FITC (Catalog #60030FI). (B) Flow cytometry analysis of C57BL/6 mouse splenocytes labeled with Anti-Mouse CD49b Antibody, Clone DX5, APC (Catalog #60020AZ) and Anti-Mouse CD45 Antibody, Clone 30-F11, FITC.
Figure 4. Data for Biotin-Conjugated
(A) Flow cytometry analysis of C57BL/6 mouse splenocytes labeled with Rat IgM, kappa Isotype Control Antibody, Clone RTK2118, Biotin followed by streptavidin (SAV) APC and Anti-Mouse CD45 Antibody, Clone 30-F11, FITC (Catalog #60030FI). (B) Flow cytometry analysis of C57BL/6 mouse splenocytes labeled with Anti-Mouse CD49b Antibody, Clone DX5, Biotin (Catalog #60020BT) followed by SAV APC and Anti-Mouse CD45 Antibody, Clone 30-F11, FITC.
Figure 5. Data for FITC-Conjugated
(A) Flow cytometry analysis of C57BL/6 mouse splenocytes labeled with Rat IgM, kappa Isotype Control Antibody, Clone RTK2118, FITC and Anti-Mouse CD45 Antibody, Clone 30-F11, PE. (B) Flow cytometry analysis of C57BL/6 mouse splenocytes labeled with Anti-Mouse CD49b Antibody, Clone DX5, FITC (Catalog #60020FI) and Anti-Mouse CD45 Antibody, Clone 30-F11, PE.
Figure 6. Data for PE-Conjugated
(A) Flow cytometry analysis of C57BL/6 mouse splenocytes labeled with Rat IgM, kappa Isotype Control Antibody, Clone RTK2118, PE and Anti-Mouse CD45 Antibody, Clone 30-F11, APC (Catalog #60030AZ). (B) Flow cytometry analysis of C57BL/6 mouse splenocytes labeled with Anti-Mouse CD49b Antibody, Clone DX5, PE (Catalog #60020PE) and Anti-Mouse CD45 Antibody, Clone 30-F11, APC. (C) Human ES cells were cultured in mTeSR™1 on Corning® Matrigel®-coated glass slides, then fixed and stained with Anti-Mouse SSEA-3 Antibody, Clone MC-631, PE (Catalog #60061PE). Inset shows cells labeled with Rat IgM, kappa Isotype Control Antibody, Clone RTK2118.
Protocols and Documentation
Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.
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Legal Statement: Alexa Fluor is a registered trademark of Life Technologies Corporation. Antibodies conjugated to Alexa Fluor® are licensed for internal research use only and sale is expressly conditioned on the buyer not using the antibody for manufacturing, performing a service or medical test, or otherwise generating revenue. For use other than research, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.com. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.
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