"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."
Janet Duerr, Ohio UniversityRating: 5.0
Jackson ImmunoResearch AffiniPure™ Anti-Rabbit secondary antibodies are affinity purified polyclonal antibodies with specificity for rabbit immunoglobulins.
We offer Anti-Rabbit secondary antibodies in a range of host species including Alpaca, Donkey, Goat, Rabbit, Rat and Sheep. JIR secondary antibodies are available as whole IgG and F(ab')2 fragment formats. Fab fragment formats are also available and can be an elegant component of multiple blocking and labeling protocols.
They are available with with specificity for whole Ig, Fcγ, and F(ab')2 domains, among other specificities.
Affinity-purified antibodies are isolated from antisera by immunoaffinity chromatography using target antigens coupled to agarose beads. A proprietary elution process is used to dissociate antibodies from the antigen. Unconjugated affinity-purified antibodies are supplied sterile filtered in phosphate buffer without stabilizers or preservatives. Conjugated affinity-purified antibodies are freeze-dried in phosphate buffer with stabilizers and sodium azide, with the exception of horseradish peroxidase conjugates, which do not contain a preservative.
Secondary antibodies against one species are likely to cross-react with other species unless they have been specifically adsorbed against the other species. Antibodies with "(min X ... Sr Prot)" in the description have been tested and/or adsorbed against IgG and/or serum proteins of those species indicated in the parentheses. They are recommended when the presence of immunoglobulins from other species may lead to interfering cross-reactivities.
“When labeling for more than one target with antibodies from closely related species, you really need to certain your secondary antibody is specific!”
Read on to find out how Cardiac researcher Elisabeth Ehler uses Jackson ImmunoResearch cross-adsorbed (min X) secondary antibodies to multiple label with primary antibodies derived from closely related host species – enabling her to differentiate cell types in primary cardiac cell cultures.
Primary cultures of neonatal rat cardiomyocytes stained with Cy3-AffiniPure Goat Anti-Mouse IgG (H+L) (115-165-146) and Cy2-AffiniPure Goat Anti-Rabbit IgG (H+L) (111-225-144) secondary antibodies together with DAPI counterstain. Image courtesy of Elisabeth Ehler, KCL’
Alexa Fluor® fluorescent dyes are widely recognized as superior fluorescent dyes, respected for their brightness and photostability. They are suitable for a variety of applications including confocal microscopy, flow cytometry, fluorescent Western blotting, and fluorescent ELISA. Alexa Fluor conjugated secondary antibodies can be used exclusively or in combination with other fluorescent probes to generate an optimal multiplex dye panel for your experimental variables.
JIR provides Alexa Fluor 488, 594, 647, 680, 790 conjugates.
Antibodies conjugated with far-red- and Infrared-emitting dyes are more sensitive than those with dyes emitting visible light due to low fluorescence quenching of the conjugates, high extinction coefficients of the dyes, and low background autofluorescence. The increased brightness allows for a wider range of immunofluorescence detection and imaging modalities. Far-red and Infrared dye conjugates can be used for higher sensitivity Western blots, quantitative Western blots, in-gel Western blots, microWestern arrays, in-cell Western arrays, on-cell Western arrays, tissue section imaging, small animal whole body imaging, and other techniques that require the brightest dyes.
Western blot using Alexa Fluor® 680 far-red dye and Alexa Fluor® 790 infrared dye. Alexa Fluor® 790-goat anti-mouse IgG, Fcγ Subclass 1 specific (min X Hu, Bov, Rb Sr Prot, 115-655-205)(green) shows heavy chains and Alexa Fluor® 680-goat anti-mouse IgG, light chain specific (min X Bov, Gt, Hrs, Hu, Rb, Rat, Shp Ig, 115-625-174)(red) shows light chains. Imaged using LiCor Odyssey imager.
