Immunofluorescence analysis of Chicken anti-Rat IgG (H+L) Secondary Antibody, Alexa Fluor 594 conjugate 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 594 conjugate (Product # A21471) 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® 594|
|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 *|
|Flow Cytometry (Flow)||1:500|
|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.
|Miscellaneous PubMed (MISC)||See 3 publications below|
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.
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.
|Not Applicable||Not Cited||Targeting the PyMT Oncogene to Diverse Mammary Cell Populations Enhances Tumor Heterogeneity and Generates Rare Breast Cancer Subtypes.||Smith BA,Shelton DN,Kieffer C,Milash B,Usary J,Perou CM,Bernard PS,Welm BE||Genes and cancer (3:550)||2012|
|Not Applicable||Not Cited||Cells derived from young bone marrow alleviate renal aging.||Yang HC,Rossini M,Ma LJ,Zuo Y,Ma J,Fogo AB||Journal of the American Society of Nephrology : JASN (22:2028)||2011|
|Not Applicable||Not Cited||Acid sphingomyelinase involvement in tumor necrosis factor alpha-regulated vascular and steroid disruption during luteolysis in vivo.||Henkes LE,Sullivan BT,Lynch MP,Kolesnick R,Arsenault D,Puder M,Davis JS,Rueda BR||Proceedings of the National Academy of Sciences of the United States of America (105:7670)||2008|