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Immunofluorescence analysis of F(ab')2-Goat anti-Mouse IgG (H+L) Secondary Antibody, Alexa Fluor 546 conjugate (Product # A11018) was performed using HeLa cells stained with alpha Tubulin (236-10501) Mouse Monoclonal Antibody (Product # A11126). 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 mouse primary antibody (1:250 dilution) for 3 hours at room temperature. F(ab')2-Goat anti-Mouse IgG (H+L) Secondary Antibody, Alexa Fluor 546 conjugate was used at concentration of 2 µg/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: green). Nuclei (Panel b: blue) were stained with DAPI in SlowFade® Gold Antifade Mountant (Product # S36938). F-actin was stained with Alexa Fluor® 488 Phalloidin (A12379, 1:300) (Panel c: red). 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||Mouse|
|Published species reactivity||Not Applicable|
|Host / Isotype||Goat / IgG|
|Immunogen||Gamma Immunoglobins Heavy and Light chains|
|Conjugate||Alexa Fluor® 546|
|Storage buffer||PBS, pH 7.5|
|Contains||5mM sodium azide|
|Storage Conditions||4° C, store in dark|
|Cross Adsorption||Against human IgG and serum|
|Antibody Form||F(ab')2 Fragment|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1-10 µg/mL|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||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 5 publications below|
Anti-Mouse secondary antibodies are affinity-purified antibodies with well-characterized specificity for mouse 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||Hepatocyte growth factor inhibits lipopolysaccharide-induced oxidative stress via epithelial growth factor receptor degradation.||Shimizu K,Taniyama Y,Sanada F,Azuma J,Iwabayashi M,Iekushi K,Rakugi H,Morishita R||Arteriosclerosis, thrombosis, and vascular biology (32:2687)||2012|
|Not Applicable||Not Cited||
Chemokine-like receptor 1 regulates skeletal muscle cell myogenesis.
A-11018 was used in immunohistochemistry - paraffin section to study the contribution of chemokine-like receptor-1 in skeletal muscles
|Issa ME,Muruganandan S,Ernst MC,Parlee SD,Zabel BA,Butcher EC,Sinal CJ,Goralski KB||American journal of physiology. Cell physiology (302:C1621)||2012|
|Not Applicable||Not Cited||On-off system for PI3-kinase-Akt signaling through S-nitrosylation of phosphatase with sequence homology to tensin (PTEN).||Numajiri N,Takasawa K,Nishiya T,Tanaka H,Ohno K,Hayakawa W,Asada M,Matsuda H,Azumi K,Kamata H,Nakamura T,Hara H,Minami M,Lipton SA,Uehara T||Proceedings of the National Academy of Sciences of the United States of America (108:10349)||2011|
|Not Applicable||Not Cited||ADP-ribosylation factor-like GTPase ARFRP1 is required for trans-Golgi to plasma membrane trafficking of E-cadherin.||Zahn C,Jaschke A,Weiske J,Hommel A,Hesse D,Augustin R,Lu L,Hong W,Florian S,Scheepers A,Joost HG,Huber O,Schürmann A||The Journal of biological chemistry (283:27179)||2008|
|Not Applicable||Not Cited||Crucial role of two potential cytosolic regions of Nox2, 191TSSTKTIRRS200 and 484DESQANHFAVHHDEEKD500, on NADPH oxidase activation.||Li XJ,Grunwald D,Mathieu J,Morel F,Stasia MJ||The Journal of biological chemistry (280:14962)||2005|