Hrp conjugated secondaries

Manufactured by Bio-Rad

HRP-conjugated secondaries are antibodies that are chemically conjugated with the enzyme Horseradish Peroxidase (HRP). These secondary antibodies are designed to bind to primary antibodies, enabling the detection and visualization of target proteins in various immunoassay techniques such as Western blotting, ELISA, and immunohistochemistry.

Automatically generated - may contain errors

Lab products found in correlation

Mouse magnetic luminex assay, by R&D Systems (1 mentions) Edta free protease and phosphatase inhibitor cocktails, by Roche (1 mentions) Odyssey, by LI COR (1 mentions) Bio plex 200, by Bio-Rad (1 mentions) Enhanced chemi luminescence (ecl), by Bio-Rad (1 mentions) Western blotting system, by Bio-Rad (1 mentions) Chemidoc imager, by Bio-Rad (1 mentions) Protease inhibitor cocktail, by Merck Group (1 mentions) Pvdf membrane, by Bio-Rad (1 mentions)

3 protocols using hrp conjugated secondaries

Immunoblotting and Luminex Assay Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell lysates (50 mM HEPES [pH 7.4], 150 mM NaCl, and 1% Triton X-100) and freshly added EDTA-free protease and phosphatase inhibitor cocktails (Roche) were immunoblotted with primary antibodies overnight at 4 °C. HRP-conjugated secondaries and ECL from BioRAD was used for chemiluminescence detection, while Dylight secondary antibodies were used for infrared detection on LI-COR Odyssey and using LI-COR Image Studio software or Image Studio Light for analysis and band quantification. Fluorescent images were saved in grey scale and therefore displayed as such. Uncropped and unprocessed blot images with molecular weight are available in the Source Data File.
For Luminex assays, samples were prepared using a customized Mouse Magnetic Luminex Assay (R&D Systems) following manufacturer’s protocol and using BioRad BioPlex 200 apparatus and software.

Blanc R.S., Kallenbach J.G., Bachman J.F., Mitchell A., Paris N.D, & Chakkalakal J.V. (2020). Inhibition of inflammatory CCR2 signaling promotes aged muscle regeneration and strength recovery after injury. Nature Communications, 11, 4167.

+ Open protocol

+ Expand

Western Blotting Analysis of Signaling Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Following boiling or elution in Laemelli Sample Buffer lysates were run on a complete Bio-Rad Western Blotting System. Samples were electrophoretically separated on Bio-Rad 10% Tris-TGX PreCast Gels and blotted onto nitrocellulose membranes using Bio-Rad Trans Turbo Dry transfer and buffer. All blots were blocked using 5% BSA. The following is a list of all primary antibodies used. Phosphorylated-Erk5 (T218/Y220) (1:200, Cell Signaling), ERK5 (1:1000, Cell Signaling), TrkB (1:1000, EMD Millipore). Immunoreactivity was measured using Bio-Rad HRP conjugated secondaries. Blots were developed on a Bio-Rad Digital Detection System running ImageLab software, using Millipore Crescendo chemiluminescence development reagent. ImageJ was used for band densitometry analysis. ImageJ quantification metrics and western blots from the same lysate were ensured to be reasonably reproducible, and all experiments were reproduced using completely independent cultures from different E18 litters.

Carlos A.J., Tong L., Prieto G.A, & Cotman C.W. (2017). IL-1β impairs retrograde flow of BDNF signaling by attenuating endosome trafficking. Journal of Neuroinflammation, 14, 29.

+ Open protocol

+ Expand

Protein Isolation from Nasal Tissue

Check if the same lab product or an alternative is used in the 5 most similar protocols

For protein isolation, nasal turbinate and septum mucosal tissue was
homogenized in ice-cold RIPA buffer containing protease inhibitor cocktail
(Sigma). Following a 30 minute incubation on ice, vortexing every 10 minutes,
the cell lysates were centrifuged at 14,000 x g for 15 minutes, and the
supernatants were collected and stored at −20C. Total protein
concentration in each sample was quantified by Bradford assay to ensure equal
protein loading of 50 mg per lane. Samples were diluted in Laemmli buffer and
heated to 100 °C for 5 minutes, analyzed by 10% Bis-Tris
polyacrylamide gel electrophoresis, and transferred to PVDF membranes (BioRad,
Hercules, CA). Membranes were blocked in 5% non-fat dried milk in Tris
buffered saline (TBS) with 0.1% Tween-20. Immunoblot analysis was
performed using mouse antibodies against SCF, control rabbit polyclonal antibody
against GAPDH (1:1000; Santa Cruz Biotechnology), and HRP-conjugated secondaries
(BioRad). Antibodies were diluted in 5% BSA in TBS with 0.1%
Tween-20. Membranes were visualized by chemiluminescence (Pierce SuperSignal) on
a ChemiDoc imager (BioRad).

Goldstein B.J., Goss G., Hatzistergos K.E., Rangel E.B., Seidler B., Saur D, & Hare J.M. (2014). Adult c-Kit (+) progenitor cells are necessary for maintenance and regeneration of olfactory neurons. The Journal of comparative neurology, 523(1), 15-31.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!