Immunofluorescence analysis of Chicken anti-Rat IgG (H+L) Secondary Antibody, Alexa Fluor® 647 was performed using A549 cells stained with alpha Tubulin (YL1/2) Rat Monoclonal Antibody (Product # MA1-80017). The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, blocked with 1% BSA for 1 hour and labeled with 2µg/ml Rat primary antibody for 3 hours at room temperature. Chicken anti-Rat IgG (H+L) Secondary Antibody, Alexa Fluor® 647 (Product # A21472) was used at a concentration of 2µg/ml in phosphate buffered saline containing 0.2 % BSA for 45 minutes at room temperature, for detection of alpha Tubulin in the cytoplasm (Panel a: red). Nuclei (Panel b: blue) were stained with DAPI in SlowFade® Gold Antifade Mountant (Product # S36938). F-actin was stained with Alexa Fluor® 488 Phalloidin (Product # A12379), 1:300) (Panel c: green). Panel d represents the composite image. No nonspecific staining was observed with the secondary antibody alone (panel f), or with an isotype control (panel e). The images were captured at 60X magnification.
|Tested species reactivity||Rat|
|Published species reactivity||Not Applicable|
|Host / Isotype||Chicken / IgY|
|Immunogen||Gamma Immunoglobins Heavy and Light chains|
|Conjugate||Alexa Fluor® 647|
|Storage buffer||PBS, pH 7.5|
|Contains||5mM sodium azide|
|Storage Conditions||4° C, store in dark|
|Cross Adsorption||Against human and rabbit IgG prior to conjugation|
|Antibody Form||Whole Antibody|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (IHC)||1-10 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Chicken anti-rat antibodies react with IgG heavy chains and all classes of immunoglobulin light chains from rat. Chicken secondary antibodies have gained popularity because they demonstrate a lower level of nonspecific binding. Chicken antibodies lack a classic “Fc” domain and will not bind to protein A or protein G, nor will they bind to mammalian IgG Fc receptors. Flourescence of this long-wavelength Alexa Fluor dye is not visible by looking through a conventional fluorescence microscope.
Anti-Rat secondary antibodies are affinity-purified antibodies with well-characterized specificity for rat immunoglobulins and are useful in the detection, sorting or purification of its specified target. Secondary antibodies offer increased versatility enabling users to use many detection systems (e.g. HRP, AP, fluorescence). They can also provide greater sensitivity through signal amplification as multiple secondary antibodies can bind to a single primary antibody. Most commonly, secondary antibodies are generated by immunizing the host animal with a pooled population of immunoglobulins from the target species and can be further purified and modified (i.e. immunoaffinity chromatography, antibody fragmentation, label conjugation, etc.) to generate highly specific reagents.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Hepatocellular carcinoma originates from hepatocytes and not from the progenitor/biliary compartment.
A-21472 was used in immunohistochemistry - paraffin section to elucidate the origin of hepatocellular carcinoma.
|Mu X,Español-Suñer R,Mederacke I,Affò S,Manco R,Sempoux C,Lemaigre FP,Adili A,Yuan D,Weber A,Unger K,Heikenwälder M,Leclercq IA,Schwabe RF||The Journal of clinical investigation (125:3891)||2015|
|Not Applicable||Not Cited||Mutagenesis of Ly49B reveals key structural elements required for promiscuous binding to MHC class I molecules and new insights into the molecular evolution of Ly49s.||Mickiewicz KM,Gays F,Lewis RJ,Brooks CG||Journal of immunology (Baltimore, Md. : 1950) (192:1558)||2014|
|Not Applicable||Not Cited||Visualization and identification of IL-7 producing cells in reporter mice.||Mazzucchelli RI,Warming S,Lawrence SM,Ishii M,Abshari M,Washington AV,Feigenbaum L,Warner AC,Sims DJ,Li WQ,Hixon JA,Gray DH,Rich BE,Morrow M,Anver MR,Cherry J,Naf D,Sternberg LR,McVicar DW,Farr AG,Germain RN,Rogers K,Jenkins NA,Copeland NG,Durum SK||PloS one (4:null)||2009|
|Not Applicable||Not Cited||Interleukin-6 is crucial for recall of influenza-specific memory CD4 T cells.||Longhi MP,Wright K,Lauder SN,Nowell MA,Jones GW,Godkin AJ,Jones SA,Gallimore AM||PLoS pathogens (4:null)||2008|
|Not Applicable||Not Cited||Cbfa1-independent decrease in osteoblast proliferation, osteopenia, and persistent embryonic eye vascularization in mice deficient in Lrp5, a Wnt coreceptor.||Kato M,Patel MS,Levasseur R,Lobov I,Chang BH,Glass DA,Hartmann C,Li L,Hwang TH,Brayton CF,Lang RA,Karsenty G,Chan L||The Journal of cell biology (157:303)||2002|