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FEATURED NEW PRODUCTS
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Study c-Met Activation With New Recombinant Antibodies — ABfinity™ Recombinant Rabbit Monoclonal Antibodies
what it is
how it works
what it is
A monoclonal antibody specific for tau (regardless of phosphorylation state) has been coated onto the wells of the microtiter strips provided. Samples are pipetted into these wells. During the first incubation, the tau antigen binds to the immobilized (capture) antibody. After washing, a rabbit antibody specific for tau [pT231] phosphorylation is added to the wells. During the second incubation, this antibody serves as a detection antibody by binding to the immobilized tau protein captured during the first incubation. After removal of excess detection antibody, a horseradish peroxidase–labeled anti-rabbit IgG (anti-rabbit IgG HRP) is added. This binds to the detection antibody to complete the four-member sandwich. After washing, a substrate solution is added, which is acted upon by the bound enzyme to produce color. The intensity of this colored product is directly proportional to the concentration of tau [pT231] present.
In this study, the researchers isolated and digested the histones, yielding peptides. These peptides were then treated with phosphatase, leaving O-GlcNAc modifications intact. Then they used a product similar to the Click-iT® O-GlcNAc Enzymatic Labeling System, wherein β-1,4-galactosyltransferase (Gal-T1) transfers azido-modified galactose (GalNAz) from UDP-GalNAz to O-GlcNAc residues on target proteins in a highly specific reaction. For detection, the researchers used a product similar to the Click-iT® Biotin Protein Analysis Detection Kit followed by mass spectrometry to detect the peptides and thus the exact O-GlcNAc modification sites.
This study illustrates the specificity and precision of click chemistry used to study the elusive sites of O-GlcNAc modifications. The click reaction is highly specific and the reaction product contains an irreversible covalent bond, making the technique ideal for accurate downstream identification with mass spectrometry.
Click-iT® Glycoprotein Detection. (A) Enzymatic labeling of an O-GlcNAc–modified protein with UDP-GalNAz and Gal-T1. (B) Click-iT® azide/alkyne reaction.
- Learn More About Click-iT® Tools for Posttranslational Modification Analysis
|Click-iT® O-GlcNAc Enzymatic Labeling System, for N- or O-linked GlcNAc glycoproteins||10 labelings||C33368|
|Click-iT® Biotin Protein Analysis Detection Kit||10 reactions||C33372|
|Click-iT® Tetramethylrhodamine (TAMRA) Protein Analysis Detection Kit, UV/532 nm excitation||10 reactions||C33370|
|Click-iT® Dapoxyl® Protein Analysis Detection Kit, for UV excitation||10 reactions||C33371|
|Click-iT® O-GlcNAc peptide and phosphopeptide LC/MS standards||1 set (5 nmol each)||C33373|
|Click-iT® O-GlcNAc peptide LC/MS standard||5 nmol||C33374|
Fluorescent proteins are highly versatile biological markers for monitoring physiological processes, visualizing protein localization, detecting protein expression, and purifying proteins. To complement these powerful tools, we offer anti-GFP and anti-RFP antibodies and antibody conjugates for a wide variety of applications, including imaging, western blotting, immunoprecipitation, and flow cytometry.
All six of our anti-GFP antibodies are suited for detection of native GFP, GFP variants, and most GFP fusion proteins. The affinity-purified anti-RFP antibody is used to detect native TagRFP and most fusion proteins derived from Entacmaea quadricolor. The RFP proteins derived from E. quadricolor have 2.8 times the quantum yield of mCherry, and more importantly, they remain as monomers, making it more likely that the fusion protein will function properly in the cell .
- Learn More About GFP Antibodies and Antibody Conjugates
- Read the BioProbes Article on GFP and RFP Antibodies
|Anti-GFP ABfinity™ Recombinant Rabbit Monoclonal Antibody used for immunocytochemistry. Left: U2OS cells expressing CellLight® ER-GFP were incubated with the anti-GFP ABfinity™ Recombinant Rabbit Monoclonal Antibody. Center: Cells were formaldehyde-fixed, permeabilized, and blocked in 1% BSA, then incubated with primary antibody at 1 μg/mL, followed by Alexa Fluor® 647 goat anti–rabbit IgG conjugate. Right: The merged yellow signal indicates colocalization of GFP fluorescence and the detection antibody. Nuclei were stained with Hoechst 33342.|
|Anti-GFP, rabbit serum (polyclonal)||ICC, IHC, IP, WB||A6455|
|Anti-GFP, chicken IgY fraction||ICC, WB||A10262|
|Anti-GFP, rabbit IgG fraction||ICC, IHC, IP, WB||A11122|
|Anti-GFP, mouse IgG2a||ICC, IHC, IP||A11120|
|Anti-GFP, mouse IgG1||ICC||A11121|
|Anti-GFP, rabbit monoclonal||ICC, IHC, IP||G10362|
|Anti-RFP, rabbit IgG polyclonal||ICC, IHC, IP, WB||R10367|
|Anti-GFP, rabbit IgG fraction, Alexa Fluor® 488 conjugate||ICC, IHC||A21311|
|Anti-GFP, rabbit IgG fraction, Alexa Fluor® 555 conjugate||ICC, IHC||A31851|
|Anti-GFP, rabbit IgG fraction, Alexa Fluor® 594 conjugate||ICC, IHC||A21312|
|Anti-GFP, rabbit IgG fraction, Alexa Fluor® 647 conjugate||ICC, IHC||A31852|
|*ICC = immunocytochemistry; IHC = cryosection immunohistochemistry; IP = immunoprecipitation; WB = western blot. |
Label. Detect. Discover.
Molecular Probes® labeling and detection solutions are designed and supported by scientists like you. At our new web page you'll find scientist interviews, stories of discovery, technical tips, and more to help inspire and advance your research.
Cells were labeled with mouse anti-giantin and rabbit anti-tubulin primary antibodies, which were detected with Alexa Fluor® 488 donkey anti–rabbit IgG (tubulin, green) and Texas Red® goat anti–mouse IgG (Golgi, red). The nucleus was stained with Hoechst 33342. Image contributed by Michael W. Davidson, Florida State University.
Imaging the Golgi Complex and Tubulin in a Rat Kidney Epithelial Cell.
|Alexa Fluor® 488 donkey anti–rabbit IgG (H+L), 2 mg/mL||0.5 mL||A21206|
|Texas Red® goat anti–mouse IgG (H+L), 2 mg/mL||0.5 mL||T862|
|Hoechst 33342, trihydrochloride, trihydrate, 10 mg/mL solution in water||10 mL||H3570|
Hemsley AL, Hernandez D, Mason C et al. (2011) Cell Health and Cytoskeleton 3:23–34.
Recent studies have revealed that embryonic stem cell (ESC) proliferation and differentiation can be influenced by exogenous mechanical forces. To further investigate the effect of these forces on mouse ESCs (mESCs), Hemsley et al. combined atomic force microscopy with laser scanning confocal microscopy to characterize morphological and biochemical responses of mESCs to precisely delivered nanomechanical forces. The researchers identified two morphologically distinct subpopulations of mESCs: round and flattened. Using phalloidin conjugated to Alexa Fluor® 546 or Alexa Fluor® 488, immunofluorescence imaging of the cytoskeleton revealed that round cells, but not flattened cells, exhibited blebbing in response to mechanically induced forces. Flattened cells were characterized by a more highly developed cytoskeleton and a stronger mechanical link between the plasma membrane and cytoskeleton. These results suggest that mechanosensitivity of ESCs at the earliest stages of differentiation may play an important role in ESC biology.
- View the Bibliography Reference
- Learn More About
Phalloidin and Phalloidin Conjugates for Staining Actin
The Molecular Probes® Handbook
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