Fluorescent TrueBlot®


TrueBlot_Banner_IF 


Rockland’s Fluorescent TrueBlot® monoclonal secondary antibodies combine the power and specificity of our original HRP-conjugated TrueBlot® monoclonal secondary antibodies with the versatility of fluorescent and near-infrared (IR) dyes. Conjugation of highly-optimized TrueBlot® secondary antibodies to an assortment of fluorescent labels results in multi-purpose reporter molecules that can be used in a variety of immunoassays. Available in Fluorescein (FITC), Dylight™ 680, or Dylight™ 800 conjugated forms targeting mouse or rabbit species, Fluorescent TrueBlot® secondary antibodies provide excellent sensitivity and are compatible with most common imaging systems. Fluorescent TrueBlot® secondary antibodies can be used to detect protein targets in a variety of Western blot, IP-Western blot, and indirect immunofluorescence (IF) expeiments.

 

The full spectrum of Fluorescent TrueBlot® secondary antibodies allows for their use in multiplex assays, where multiple targets can be detected and imaged simultaneously, resulting in distinct protein bands labeled by a unique identifying color. Simultaneous detection cuts time, ensures target molecule identification, and eliminates the need for stripping and reprobing the blot. The combination of Dylight™ 680 and Dylight™ 800 dyes with near-IR imaging, yields higher signal-to-noise ratios, as background autofluorescence in the near-IR region is relatively low.



How do Fluorescent TrueBlot® Secondary Antibodies Work?


Western blots of immunoprecipitated samples can frequently result in the detection of undesirable background signals, specifically when the same antibody is used as the capture antibody in immunoprecipitation (IP) and the primary antibody in Western blot. The final steps in a conventional IP experiment typically employ a final heating step in the presence of reducing/denaturing reagents when the desired downstream application is SDS-PAGE and Western blot. When the eluted IP sample is separated by SDS-PAGE and followed by detection with conventional secondary antibodies in a Western blot, both the heavy and light chain of the IP capture antibody can be detected and obscure your antigen of interest. TrueBlot® secondary antibodies solve this problem by only recognizing primary antibodies in their native (non-reduced) state, eliminating interference from heavy and light chain bands and allowing you to obtain accurate, publication-quality Western blots.



Use in Western Blot and IP-Western Blot


The monoclonal nature of TrueBlot® secondary antibodies limits non-specific binding and provides reliable reproducibility. Moreover, the unique ability of TrueBlot® to differentiate between the reduced and non-reduced forms of IgG, make TrueBlot® secondary antibodies the ideal solution when performing Western blot experiments that directly follow IP (IP-Western Blot), as TrueBlot® secondary antibodies reduce interference by the ~55 kDa heavy and ~23 kDa light chains of the immunoprecipitating antibody.

 

 

 Fluorescent TrueBlot®
Anti-Mouse IgG DyLight™ 680
18-4417-32

 

Fluorescent TrueBlot®
Anti-Rabbit IgG Fluorescein
18-0216-32

 

Fluorescent TrueBlot®
Anti-Mouse IgG Fluorescein
18-0217-32

18-4417-32 Fluorescent TrueBlot a Mouse IgG DyLight 680 Conjugated 3 WB

 

18-0216-32 Fluorescent TrueBlot a Rabbit IgG Fluorescein 3 WB

 

18-0217-32 Fluorescent TrueBlot a Mouse IgG Fluorescein WB

Unique Detection Properties: Fluorescent TrueBlot® secondary antibodies detect the non-reduced form of IgG (Lane 1; all images) over the reduced form of IgG (Lane 2; all images). Lane M (all images) represents the molecular weight marker. The expected band for non-reduced IgG is ~160 kDa.
Note: Observed non-reduced bands migrate at a slightly higher molecular weight.  
 

 


Pair Fluorescent TrueBlot® secondary antibodies with the appropriate species-specific TrueBlot® Ig IP agarose or magnetic beads to purify and detect even the most challenging protein targets, including proteins present in low abundance due to poor or minimal expression, protein targets that undergo post-translational modifications (i.e. phosphorylation, acetylation), and protein-protein interactions.

 



Use in Immunofluorescence (IF)


Although ideal as a reporter molecule in IP-Western Blot experiments, Fluorescent TrueBlot® secondary antibodies are versatile and can be used to analyze the distribution of proteins, protein modifications, and other antigen targets in indirect immunofluorescence microscopy providing high-quality images suitable for publication.

 

   ZO-1    Alpha-tubulin  
  Immunofluorescence microscopy of anti-ZO-1 primary antibody (600-401-GU7) in Caco-2 cells using FITC-conjugated Fluorescent TrueBlot® anti-rabbit IgG for detection.      Immunofluorescence microscopy of anti-α-tubulin primary antibody (200-301-880) in A431 cells using DyLight™ 680-conjugated TrueBlot® anti-mouse Ig for detection.     

 

 

Fluorescent TrueBlot® Products


Product Catalog # Applications Size (μL)
Fluorescent TrueBlot®: Anti-Rabbit IgG Fluorescein 18-0216-32 IF, IP, WB 100
Fluorescent TrueBlot®: Anti-Mouse IgG Fluorescein  18-0217-32 IF, IP, WB 100
Fluorescent TrueBlot®: Anti-Rabbit IgG DyLight™ 680 18-4416-32 IF, IP, WB 100
Fluorescent TrueBlot®: Anti-Mouse IgG DyLight™ 680 18-4417-32 IF, IP, WB 100
Fluorescent TrueBlot®: Anti-Rabbit IgG DyLight™ 800 18-4516-32 IP, WB 100
Fluorescent TrueBlot®: Anti-Mouse IgG DyLight™ 800 18-4517-32 IP, WB 100


 

TrueBlot Reagents Button 2 

 

  Related
Fluorescent Dyes


Phospho-specific Antibodies


TrueBlot® for IP-Western Blot  

 

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