The largest database of trusted experimental protocols

Goat α rabbit 680 nm

Manufactured by Thermo Fisher Scientific

The Goat α-Rabbit 680 nm is a secondary antibody product designed for use in immunofluorescence and Western blot applications. It is a polyclonal antibody raised in goats and specifically recognizes rabbit primary antibodies. The antibody is conjugated to a fluorescent dye that emits light at a wavelength of 680 nm when excited.

Automatically generated - may contain errors

3 protocols using goat α rabbit 680 nm

1

Whole Cell Bacterial Lysate Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols
Whole cell bacterial lysates were prepared for Western blot analysis as follows. Bacteria were harvested from overnight growth on CFA agar with or without 200 µM DFOM in PBS, and the OD600 was used to normalize to the samples for bacterial numbers (55 (link)). The normalized suspensions were then mixed 1:1 with 2× Laemmli Sample Buffer (Bio-Rad). Proteins were separated on a 12% Mini-Protean TGX Precast Gel (Bio-Rad) and transferred to PVDF membrane. The membrane was blocked in a 10% (wt/vol) nonfat milk buffer in PBS and incubated with absorbed polyclonal Rabbit α-CS14 (Rockland Immunochemicals). The secondary antibody was Goat α-Rabbit 680 nm (Invitrogen), and proteins were visualized using the LI-COR Odyssey Laser Scanner. Additional Western blots of CFA agar with or without 200 µM DFOM grown strains were performed and stained with Rabbit anti-DnaK (Invitrogen) and Goat α-Rabbit 680 nm (Invitrogen) antibodies to confirm equal loading.
+ Open protocol
+ Expand
2

Whole Cell Bacterial Lysates for Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols
Whole cell bacterial lysates were prepared for western blot analysis as follows. Bacteria were harvested from overnight growth on CFA agar plates or in Terrific broth static culture into PBS, and the OD600 was used to normalize the samples for bacterial numbers. The normalized suspensions were then mixed 1:1 with 2x Laemmli Sample Buffer (Bio-Rad). Proteins were separated on a 12% Mini-Protean TGX Precast Gel (Bio-Rad) and transferred to PVDF membrane. The membrane was blocked in a 10% (w/v) nonfat milk buffer in PBS and incubated with absorbed polyclonal Rabbit α-CFA/I (Rockland Immunochemicals) or polyclonal Rabbit α-LngA antibodies (Genscript). The secondary antibody was Goat α-Rabbit 680 nm (Invitrogen), and proteins were visualized using the LI-COR Odyssey Laser Scanner. An identical 12% Mini-Protean TGX Precast Gel (Bio-Rad) was run with each Western blot and stained with GelCode Blue Stain Reagent (Thermo Scientific) to determine the total protein in each lane. Additional Western blots of CFA agar grown and Terrific broth grown strains were performed and stained with anti-DnaK (Invitrogen) and Goat α-Rabbit 680 nm (Invitrogen) antibodies to confirm equal loading. Fiji (ImageJ) was used to quantify the amount of protein (ng) expressed by each sample within individual Western blots as compared to the protein concentration of the purified fimbrial control.
+ Open protocol
+ Expand
3

Whole Cell Bacterial Lysates for Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols
Whole cell bacterial lysates were prepared for western blot analysis as follows. Bacteria were harvested from overnight growth on agar plates into 1xDPBS (Corning, Corning, NY) and the OD600 was used to normalize to an OD of 3.0. The normalized suspensions were then mixed 1:1 with 2x Laemmli Sample Buffer (0.125 M Tris pH 6.8, 4% SDS, 20% Glycerol, 0.2% Bromophenol Blue, 10% β-mercaptoethanol). Proteins were separated on a 12% Mini-Protean TGX Precast Gel (Bio-Rad, Hercules, CA) before being transferred to a PVDF membrane. The membrane was blocked in a 10%(w/v) nonfat milk buffer in 1xDPBS and then incubated with polyclonal Rabbit α-CS6 sera (Rockland Immunochemicals, Limerick, PA) in 10% (w/v) nonfat milk buffer overnight at 4°C on an orbital shaker. Following washing, the membrane was incubated with secondary antibody Goat α-Rabbit 680 nm (Invitrogen, Carlsbad, CA) and proteins visualized using the 700 nm wavelength setting on an LI-COR Odyssey Laser Scanner.
+ 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?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!