Immunofluorescence analysis of Goat anti-Rabbit IgG (H+L) Secondary Antibody, Rhodamine Red-X was performed using HepG2 cells stained with alpha-1 antitrypsin Rabbit Polyclonal Primary Antibody (PA516661). 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 Rabbit primary antibody (1:250 dilution) for 3 hours at room temperature. Goat anti-Rabbit IgG (H+L) Secondary Antibody, Rhodamine Red-X (R6394) was used at a concentration of 4ug/ml in phosphate buffered saline containing 0.2 % BSA for 45 minutes at room temperature, for detection of alpha-1 antitrypsin in the cytoplasm (Panel a: red). Nuclei (Panel b: blue) were stained with DAPI in SlowFade® Gold Antifade Mountant (S36938). F-actin was stained with Alexa Fluor® 488 Phalloidin (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||Rabbit|
|Published species reactivity||Not Applicable|
|Host / Isotype||Goat / IgG|
|Immunogen||Gamma Immunoglobins Heavy and Light chains|
|Storage buffer||PBS, pH 7.5|
|Contains||5mM sodium azide|
|Storage Conditions||4° C, store in dark|
|Cross Adsorption||Against human IgG, human serum, mouse IgG, mouse serum and bovine serum|
|Antibody Form||Whole Antibody|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||4 µg/ml|
|Immunofluorescence (IF)||4 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.
Anti-Rabbit secondary antibodies are affinity-purified antibodies with well-characterized specificity for rabbit 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.
Sialic acid expression in the mosquito Aedes aegypti and its possible role in dengue virus-vector interactions.
R-6394 was used in immunohistochemistry to analyze sialic acid expression and its possible role in dengue virus-vector interactions in the mosquito Aedes aegypti
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|Not Applicable||Not Cited||Transgenic mice expressing a cameleon fluorescent Ca2+ indicator in astrocytes and Schwann cells allow study of glial cell Ca2+ signals in situ and in vivo.||Atkin SD,Patel S,Kocharyan A,Holtzclaw LA,Weerth SH,Schram V,Pickel J,Russell JT||Journal of neuroscience methods (181:212)||2009|
|Not Applicable||Not Cited||Mechanism of cell death caused by complex I defects in a rat dopaminergic cell line.||Marella M,Seo BB,Matsuno-Yagi A,Yagi T||The Journal of biological chemistry (282:24146)||2007|
|Not Applicable||Not Cited||Gangliosides inhibit flagellin signaling in the absence of an effect on flagellin binding to toll-like receptor 5.||West AP,Dancho BA,Mizel SB||The Journal of biological chemistry (280:9482)||2005|
|Not Applicable||Not Cited||Multi-wavelength immunoassays using surface plasmon-coupled emission.||Matveeva E,Malicka J,Gryczynski I,Gryczynski Z,Lakowicz JR||Biochemical and biophysical research communications (313:721)||2004|
|Not Applicable||Not Cited||Ggamma subunit-selective G protein beta 5 mutant defines regulators of G protein signaling protein binding requirement for nuclear localization.||Rojkova AM,Woodard GE,Huang TC,Combs CA,Zhang JH,Simonds WF||The Journal of biological chemistry (278:12507)||2003|
|Not Applicable||Not Cited||Physical and functional interaction between myeloid cell leukemia 1 protein (MCL1) and Fortilin. The potential role of MCL1 as a fortilin chaperone.||Zhang D,Li F,Weidner D,Mnjoyan ZH,Fujise K||The Journal of biological chemistry (277:37430)||2002|
|Not Applicable||Not Cited||The CRE/CREB pathway is transiently expressed in thalamic circuit development and contributes to refinement of retinogeniculate axons.||Pham TA,Rubenstein JL,Silva AJ,Storm DR,Stryker MP||Neuron (31:409)||2001|
|Not Applicable||Not Cited||The Alzheimer amyloid precursor protein (APP) and FE65, an APP-binding protein, regulate cell movement.||Sabo SL,Ikin AF,Buxbaum JD,Greengard P||The Journal of cell biology (153:1403)||2001|
|Not Applicable||Not Cited||A new focal adhesion protein that interacts with integrin-linked kinase and regulates cell adhesion and spreading.||Tu Y,Huang Y,Zhang Y,Hua Y,Wu C||The Journal of cell biology (153:585)||2001|
|Not Applicable||Not Cited||A role for actin, Cdc1p, and Myo2p in the inheritance of late Golgi elements in Saccharomyces cerevisiae.||Rossanese OW,Reinke CA,Bevis BJ,Hammond AT,Sears IB,O'Connor J,Glick BS||The Journal of cell biology (153:47)||2001|
|Not Applicable||Not Cited||Temporal and spatial distribution of activated Pak1 in fibroblasts.||Sells MA,Pfaff A,Chernoff J||The Journal of cell biology (151:1449)||2000|
|Not Applicable||Not Cited||Golgi apparatus immunolocalization of endomannosidase suggests post-endoplasmic reticulum glucose trimming: implications for quality control.||Zuber C,Spiro MJ,Guhl B,Spiro RG,Roth J||Molecular biology of the cell (11:4227)||2000|
|Not Applicable||Not Cited||Herp, a new ubiquitin-like membrane protein induced by endoplasmic reticulum stress.||Kokame K,Agarwala KL,Kato H,Miyata T||The Journal of biological chemistry (275:32846)||2000|
|Not Applicable||Not Cited||Overexpression of PAX6(5a) in lens fiber cells results in cataract and upregulation of (alpha)5(beta)1 integrin expression.||Duncan MK,Kozmik Z,Cveklova K,Piatigorsky J,Cvekl A||Journal of cell science (113 ( Pt 18):3173)||2000|
|Not Applicable||Not Cited||Identification of 12-lipoxygenase interaction with cellular proteins by yeast two-hybrid screening.||Tang K,Finley RL,Nie D,Honn KV||Biochemistry (39:3185)||2000|
|Not Applicable||Not Cited||Identification and characterization of a novel Golgi protein, golgin-67.||Jakymiw A,Raharjo E,Rattner JB,Eystathioy T,Chan EK,Fujita DJ||The Journal of biological chemistry (275:4137)||2000|
|Not Applicable||Not Cited||p21-activated kinase 1 (Pak1) regulates cell motility in mammalian fibroblasts.||Sells MA,Boyd JT,Chernoff J||The Journal of cell biology (145:837)||1999|
|Not Applicable||Not Cited||Fab1p is essential for PtdIns(3)P 5-kinase activity and the maintenance of vacuolar size and membrane homeostasis.||Gary JD,Wurmser AE,Bonangelino CJ,Weisman LS,Emr SD||The Journal of cell biology (143:65)||1998|