Immunofluorescence analysis of Goat anti-Rat IgG (H+L) Secondary Antibody, Alexa Fluor® 633 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. Goat anti-Rat IgG (H+L) Secondary Antibody, Alexa Fluor® 633 (Product # A21094) 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||Goat / IgG|
|Immunogen||Gamma Immunoglobins Heavy and Light chains|
|Conjugate||Alexa Fluor® 633|
|Storage buffer||PBS, pH 7.5|
|Contains||5mM sodium azide|
|Storage Conditions||4° C, store in dark|
|Cross Adsorption||Against mouse IgG, mouse serum and human serum 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.
|Miscellaneous PubMed (MISC)||See 7 publications below|
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||Glycoprotein B cleavage is important for murid herpesvirus 4 to infect myeloid cells.||Glauser DL,Milho R,Frederico B,May JS,Kratz AS,Gillet L,Stevenson PG||Journal of virology (87:10828)||2013|
|Not Applicable||Not Cited||Distinct roles of cadherin-6 and E-cadherin in tubulogenesis and lumen formation.||Jia L,Liu F,Hansen SH,Ter Beest MB,Zegers MM||Molecular biology of the cell (22:2031)||2011|
|Not Applicable||Not Cited||Alterations in marginal zone macrophages and marginal zone B cells in old mice.||Birjandi SZ,Ippolito JA,Ramadorai AK,Witte PL||Journal of immunology (Baltimore, Md. : 1950) (186:3441)||2011|
|Not Applicable||Not Cited||Secretagogue stimulation enhances NBCe1 (electrogenic Na(+)/HCO(3)(-) cotransporter) surface expression in murine colonic crypts.||Yu H,Riederer B,Stieger N,Boron WF,Shull GE,Manns MP,Seidler UE,Bachmann O||American journal of physiology. Gastrointestinal and liver physiology (297:G1223)||2009|
|Not Applicable||Not Cited||N-glycosylation of the I-like domain of beta1 integrin is essential for beta1 integrin expression and biological function: identification of the minimal N-glycosylation requirement for alpha5beta1.||Isaji T,Sato Y,Fukuda T,Gu J||The Journal of biological chemistry (284:12207)||2009|
|Not Applicable||Not Cited||The hyaluronan receptors CD44 and Rhamm (CD168) form complexes with ERK1,2 that sustain high basal motility in breast cancer cells.||Hamilton SR,Fard SF,Paiwand FF,Tolg C,Veiseh M,Wang C,McCarthy JB,Bissell MJ,Koropatnick J,Turley EA||The Journal of biological chemistry (282:16667)||2007|
|Not Applicable||Not Cited||AP1B sorts basolateral proteins in recycling and biosynthetic routes of MDCK cells.||Gravotta D,Deora A,Perret E,Oyanadel C,Soza A,Schreiner R,Gonzalez A,Rodriguez-Boulan E||Proceedings of the National Academy of Sciences of the United States of America (104:1564)||2007|