Kaede [2F4] antibody

Overview


	Antigen:	Kaede [2F4]
	Antibody Type:	Monoclonal
	Application:	Immunoprecipitation (IP)
	Host:	Mouse
	Quantity:	10 µl (1 mg/mL)
	Price:	66,00 € (Plus shipping costs and VAT)
	Order number:	ABIN131951
	Availability:	7 to 10 days until shipment

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Additional Information: Kaede [2F4] antibody

Antigen	Kaede [2F4]
Immunogen	This antibody was purified from hybridoma (clone 2F4) supernatant using protein A column. This hybridoma was established by fusion of mouse myeloma cell P3-U1 with C3H mouse lymphocyte immunized with recombinant Kaede.
Antibody Type	Monoclonal
Isotype	IgG1. »Matching secondary antibodies
Description	CoralHue™ Kaede protein emits green fluorescence that can be irreversibly converted to red. The red fluorescence is comparable in intensity to the green and is stable under usual aerobic conditions. The green-to-red conversion is highly sensitive to irradiation with UV or violet light (350-410 nm). Maximal illumination results in a 2,000-fold increase in the ratio of red-to-green signal. The excitation lights used to elicit red and green fluorescence do not induce the photoconversion. CoralHue™ Kaede provides a simple and powerful technique for regional optical marking.
Clone	2F4
Host	Mouse
Specificity	This antibody clearly immuno- precipitates CoralHue™ Kaede. The antibody also exhibits slight cross-reactivity with CoralHue™ Midoriishi-Cyan and CoralHue™ Dronpa.

Application Details: Kaede [2F4] antibody

Application	Immunoprecipitation (IP)
Application Notes	Immunoprecipitation 1) Add primary antibody as suggested in the Applications into 30 µL of 50% protein A agarose beads resuspended in the cold Lysis buffer (50 mM Tris-HCl pH 7.2, 250 mM NaCl, 0.1% NP-40, 2 mM EDTA, 10% glycerol). Mix well and incubate with gentle agitation for 60 minutes at 4 o C. 2) Wash the beads 3-5 times with the cold Lysis buffer (centrifuge the tube at 2,500 x g for 10 seconds). 3) Add 100 µL of the recombinant protein. Mix well and incubate with gentle agitation for 60 minutes at 4 o C. 4) Wash the beads 3-5 times with the cold Lysis buffer (centrifuge the tube at 2,500 x g for 10 seconds). Im m unoprecipita tio n o f K a e d e fro m E.coli w ith norm al m ouse IgG1 (1) or M 106-3 (2). After im m unoprecipitate d w ith th e a n tibody, im m unocom plex w as resolved on SDS-PA GE and im m unoblotted with P M 0 0 2 (A n ti-His-Tag polyclonal a n tibody). IgG Heavy chain Kaede IgG Light Chain 66 45 30 20 12 kD a Im m unoprecipita tio n o f K a e d e fro m E.coli w ith norm al m ouse IgG1 (1) or M 106-3 (2). After im m unoprecipitate d w ith th e a n tibody, im m unocom plex w as resolved on SDS-PA GE and im m unoblotted with P M 0 0 2 (A n ti-His-Tag polyclonal a n tibody). IgG Heavy chain Kaede IgG Light Chain 66 45 30 20 12 kD a IgG Heavy chain IgG Heavy chain Kaede Kaede IgG Light Chain IgG Light Chain 66 45 30 20 12 kD a 66 45 30 20 12 kD a 5) Resuspend the beads in 20 µL of Laemmli’s sample buffer and boil the samples for 2 minutes and centrifuge. Load 10 µL of the sample per lane in a 1 mm thick SDS-polyacrylamide gel for electrophoresis. 6) Blot the protein to a polyvinylidene difluoride (PVDF) membrane at 10V for 45 minutes in a semi-dry transfer system (Transfer Buffer - 25 mM Tris, 190 mM glycine, 20% MeOH). See the manufacture's manual for precise transfer procedure. 7) To reduce nonspecific binding, soak the membrane in 100% Block Ace for 1 hour at room temperature, or overnight at 4 o C. 8) Incubate the membrane with the 1:1,000 anti-His-Tag antibody (MBL, code no.PM002) diluted with 10% Block Ace (in PBS, pH 7.2) for 1 hour at room temperature. 9) Wash the membrane with PBS (5 minutes x 6 times). 10) Incubate the membrane with the 1:10,000 HRP-conjugated anti-rabbit IgG (BioRad, code no. 170-6515) diluted with 1% BSA (in PBS, pH 7.2) for 1 hour at room temperature. 11) Wipe excess buffer from the membrane, then incubate it with appropriate chemiluminescence reagents for 1 minute. Remove extra reagent from the membrane by dabbing with a paper towel, and seal it in plastic wrap. 12) Expose to an X-ray film in a dark room for 2 minutes. Develop the film as usual. The conditions for exposure and development may vary. 050609-1 Western blotting: Not recommended. Immunoprecipitation: 2 µg / 100 µL of the recombinant protein. Immunohistochemistry: Not tested. Immunocytochemistry: Not tested. Flow Cytometry: Not tested.
Concentration	1 mg/mL
Buffer	100 µg IgG in 100 µL PBS containing 50% glycerol, pH 7.2. Contains no preservatives.
Restrictions	For Research Use Only

Images: Kaede [2F4] antibody

Immunoprecipitation of Kaede from E.coli w ith norm al m ouse IgG 1 (1) or ABIN131950(2) . A fter im m unoprecipitated w ith the antibody, im m unocom plex w as resolved on S D S -P A G E and im m unoblotted w ith PM002 (A nti-H is-T ag polyclonal antibody).

Image for Kaede [2F4] antibody (1)

External Information: Kaede [2F4] antibody

Google Scholar	Find more references for clone 2F4 on Google Scholar™
EMBL Harvester	Search human, mouse or rat genome for antigen Kaede [2F4] (Bioinformatic Harvester (EMBL))