Western blot analysis was performed on nuclear extracts (30 µg lysate) of HeLa (Lane 1) and U-87 MG (Lane 2). The blots were probed with ABfinity™ Anti-JUNB Recombinant Rabbit Monoclonal Antibody (Product # 701702, 2 µg/ml) and detected using Donkey anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor 680 (Product # A10043) at concentrations 0.04 µg/ml (Fig. 1), 0.1 µg/ml (Fig. 2) and 0.2 µg/ml (Fig. 3). A 42 kDa band corresponding to JUNB was observed. Known quantity of protein samples were electrophoresed using Novex® NuPAGE®12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary antibody after blocking with 5 % skimmed milk. Fluorescent detection was performed using the Odyssey® Fc imaging system (Li-cor Biosciences).
|Tested species reactivity||Rabbit|
|Published species reactivity||Not Applicable|
|Host / Isotype||Donkey / IgG|
|Conjugate||Alexa Fluor® 680|
|Storage buffer||PBS, pH 7.5|
|Contains||5mM sodium azide|
|Storage Conditions||4° C, store in dark|
|Antibody Form||Whole Antibody|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:1,000|
|Immunocytochemistry (ICC)||4 µg/ml|
|Immunofluorescence (IF)||4 µg/ml|
|Western Blot (WB)||0.04-0.2 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 8 publications below|
This secondary antibody is designed for fluorescent Western blot detection on various near-infrared fluorescence instruments. This antibody can be used for multi-color and multiplexing detection when using other antibodies conjugated to compatible Alexa Fluor™ dyes and wavelengths. Other applications of this antibody include immunofluorescent and fluorescent imaging applications when using instrumentation with appropriate excitation and detection capabilities. This antibody shows minimum cross-reactivity to bovine, chicken, goat, guinea pig, hamster, horse, human, rabbit, rat, and sheep serum proteins.
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.
|Not Applicable||Not Cited||Canonical and noncanonical Wnt proteins program dendritic cell responses for tolerance.||Oderup C,LaJevic M,Butcher EC||Journal of immunology (Baltimore, Md. : 1950) (190:6126)||2013|
|Not Applicable||Not Cited||Infection with Mycobacterium avium subsp. paratuberculosis results in rapid interleukin-1β release and macrophage transepithelial migration.||Lamont EA,O'Grady SM,Davis WC,Eckstein T,Sreevatsan S||Infection and immunity (80:3225)||2012|
|Not Applicable||Not Cited||Rel A/p65 is required for cytokine-induced myotube atrophy.||Yamaki T,Wu CL,Gustin M,Lim J,Jackman RW,Kandarian SC||American journal of physiology. Cell physiology (303:C135)||2012|
|Not Applicable||Not Cited||The expression level of HJURP has an independent prognostic impact and predicts the sensitivity to radiotherapy in breast cancer.||Hu Z,Huang G,Sadanandam A,Gu S,Lenburg ME,Pai M,Bayani N,Blakely EA,Gray JW,Mao JH||Breast cancer research : BCR (12:null)||2010|
|Not Applicable||Not Cited||Independent transport and sorting of functionally distinct protein families in Tetrahymena thermophila dense core secretory granules.||Rahaman A,Miao W,Turkewitz AP||Eukaryotic cell (8:1575)||2009|
|Not Applicable||Not Cited||An azido-biotin reagent for use in the isolation of protein adducts of lipid-derived electrophiles by streptavidin catch and photorelease.||Kim HY,Tallman KA,Liebler DC,Porter NA||Molecular and cellular proteomics : MCP (8:2080)||2009|
|Not Applicable||Not Cited||Extrasynaptic release of GABA by retinal dopaminergic neurons.||Hirasawa H,Puopolo M,Raviola E||Journal of neurophysiology (102:146)||2009|
|Not Applicable||Not Cited||Identification of WNT/beta-CATENIN signaling pathway components in human cumulus cells.||Wang HX,Tekpetey FR,Kidder GM||Molecular human reproduction (15:11)||2009|