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Whole IgG Affinity-Purified Secondary Antibodies

"I have used a wide variety of secondaries and Jackson ImmunoResearch has consistently been the best. The fluorophores are bright and stable and their selective (x reactivity removed) secondaries have always shown species specificity in multiple labeling."

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Mouse Subclass Specific Secondary Antibodies

Anti-Mouse IgG Subclass Specific Secondary Antibodies

Mouse IgG subclasses

Mice express four of the five available IgG subclasses making up their IgG isotype. The four IgG subclasses work in synergy as part of the mouse immune response. Mice typically encode for IgG1, IgG2b, and IgG3 and, depending on their strain, will express either IgG2a or IgG2c (Collins 2016). IgG2a, IgG2b, and IgG2c subclasses have similar functions (Collins 2016).

Mouse standing on a gloved hand

Inbred mouse strains, such as BALB/c and Swiss Webster mice, possess the Igh1-a allele, which results in the expression of IgG2a, the gene for IgG2c being absent. Whereas in mouse strains such as C57Bl/6, C57Bl/10, SJL, and NOD mice, the IgG2a gene is deleted. The possession of an Igh1-b allele results in the expression of IgG2c instead of the IgG2a subclass (Martin et al. 1998).

Several monoclonal antibodies originate from inbred stains, and some IgG2c clones have been incorrectly isotyped as IgG2a by reagents that cannot distinguish between these two subclasses. Subclass-specific antibodies from Jackson ImmunoResearch can be used to discriminate between the subclasses accurately.

Mouse strain Allele IgG Subclass
C57BI/6 Igh1-b IgG2c
C57Bl/10
SJL
NOD
BALB/c Igh1-a IgG2a
Swiss Webster

Distinguish between different mouse IgG subclasses

Jackson ImmunoResearch Anti-Mouse IgG, subclass specific antibodies offer specificity to the five individual mouse IgG subclasses. These highly specific antibodies are designed to distinguish between two or more different subclass of mouse IgG in multiple labeling experiments or for mouse IgG subclass determination.

They are minimally cross-reactive against human, bovine, and rabbit serum proteins to minimize interference from cross-reactivity with tissue immunoglobulins, adherent bovine IgG on cultured cells, and rabbit-derived primary antibodies.

Anti-Mouse IgG, subclass specific antibodies are available conjugated to Alexa Fluor® and Cyanine™ Fluorescent dyes, Biotin-SP™, and horseradish peroxidase and alkaline phosphatase reporter molecules.


The example below illustrates the performance of the Goat Anti-Mouse subclass specific antibodies in a dot blot experiment. In each case, the subclass specific antibody only recognizes its specific target in the presence of additionally blotted subclasses.

Dot blot showing the specificity of goat anti-mouse IgG, Fcγ subclass specific antibodies.
Figure 1: Dot blot showing the specificity of goat anti-mouse IgG, Fcγ subclass specific antibodies.
Separate nitrocellulose strips (rows) received 100 ng “dots” of mouse IgG and each subclass, and then were blocked with 5% (w/v) BSA in PBST. After probing with Peroxidase-conjugated goat anti-mouse subclass specific antibodies, the strips were developed with TMBM substrate from Moss, Inc. The grid of positive signals shows the specificity of each subclass directed antibody. Some subclasses are poorly represented in a total IgG pool (IgGtotal, α-IgGs 2c and 3) and thus give weak signal. Peroxidase conjugates used for probing were 115-035-205 (anti-mouse IgG1), 115-035-206 (anti-mouse IgG2a), 115-035-207 (anti-mouse IgG2b), 115-035-208 (anti-mouse IgG2c), and 115-035-209 (anti-mouse IgG3).

*Subclass specific antibodies are not necessary for general detection of mouse monoclonal antibodies in single-labeling experiments or in multiple-labeling experiments involving one mouse monoclonal and primary antibodies from other species.


Mouse Subclass Specific Secondary Antibodies


References

  • Collins, A., 2016. IgG subclass co‐expression brings harmony to the quartet model of murine IgG function. Immunology & Cell Biology, 94(10), pp.949-954.
  • Martin, R., Brady, J. and Lew, A., 1998. The need for IgG2c specific antiserum when isotyping antibodies from C57BL/6 and NOD mice. Journal of Immunological Methods, 212(2), pp.187-192.
  • Morgado, M., Cam, P., Gris-Liebe, C., Cazenave, P. and Jouvin-Marche, E., 1989. Further evidence that BALB/c and C57BL/6 gamma 2a genes originate from two distinct isotypes. The EMBO Journal, 8(11), pp.3245-3251.
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