Name:
Phone:
E-mail:
Address:


 


BirA (mutated/TurboID)

AS20 4440  | Clonality: Polyclonal |  Host: Rabbit | Reactivity: E. coli BirA (mutated/TurboID)

BirA (mutated/TurboID) in the group Tag Antibodies / GAL4 / GUS / LUC / others at Agrisera AB (Antibodies for research) (AS20 4440)

DATA SHEET IN PDF

292
Buy 2 items of this product for 218.00 €/items
Buy 3 items of this product for 198.00 €/items
How to cite this product:
Product name, number (Agrisera, Sweden)

Data sheet Product citations Protocols Add review

Product Information

Immunogen Recombinat mutated protein of the BirA protein from E.coli
Host

Rabbit

Clonality Polyclonal
Purity Affinity purified serum, in PBS pH 7.4
Format Lyophilized
Quantity 50 µg
Reconstitution For reconstitution add 50 µl, of sterile water.
Storage Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.
Tested applications Western blot (WB)
Recommended dilution 1 : 5000 (WB)
Expected | apparent MW 35 kDa

Reactivity

Confirmed reactivity TurboID
Not reactive in No confirmed exceptions from predicted reactivity are currently known

Application examples

Application examples Western blot using anti-TurboID antibodies

1 – 20 µg A. thaliana – Wt Col-0 (Negative control)
2 – 20 µg of A. thaliana expressing POI-TurboID fusion (Independent line 1)
3 – 20 µg of A. thaliana expressing POI-TurboID fusion (Independent line 2)
4 – 20 µg of A. thaliana expressing POI-TurboID fusion (Independent line 3)
5 – 10 ng of purified TurboID (Positive control)

20 µg/well of total protein were extracted from Arabidopsis thaliana leaf material in diluted HENS (25mM HEPES pH 7.7, 1mM EDTA, 2.5 % SDS) and stored at -80°C. Samples were denatured in 1x protein loading dye (0.5% Sodium dodecyl Sulfate, 0.002% Bromophenol Blue, 10% glycerol, and 50 mM Tris-HCl pH6.8) at 95°C for 5 min. Samples were separated on 4-16% gradient SDS-PAGE gel and blotted 1h to a nitrocellulose membrane (pore size of 0.45 um), using a semi-dry transfer. Blot was blocked with 5% milk in TBS-T at 4°C/ON without agitation. Blot was incubated in the primary antibody at a dilution of 1:5000 in 5% milk in TBS-T, at 4°C, ON without agitation. The antibody solution was decanted and the blot was rinsed briefly, then washed three times for 10 min in TBS-T at RT with agitation. Blot was incubated in Agrisera matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, AS09 602) diluted to 1:25 000 in TBS-T for 1h at RT with agitation. The blot was washed as above and developed for 2 min with Agrisera ECLBright. Exposure time was 172 seconds.

Western blot using anti-TurboID antibodies

1 – 1 ng of purified TurboID 2 – 5 ng of purified TurboID 3 – 10 ng of purified TurboID 4 – 25 ng of purified TurboID 5 – 50 ng of purified TurboID 6 – 75 ng of purified TurboID Mark: PageRulerTM Plus Prestained Protein Ladder; ThermoFisher Scientific; MW of TurboID = 35 kDa

1ng, 5 ng, 10 ng, 25 ng, 50 ng, and 75 ng loaded into wells 1, 2, 3, 4, 5, and 6, respectively, of purified TurboID in 1x Phosphate Buffer Saline (PBS) were stored at -80°C. Samples were denatured in 1x protein loading dye (0.5% Sodium dodecyl Sulfate, 0.002% Bromophenol Blue, 10% glycerol, and 50 mM Tris-HCl pH6.8) at 95°C for 5 min. Samples were separated on 4-16% gradient SDS-PAGE gel and blotted 1h to a nitrocellulose membrane (pore size of 0.45 um), using a semi-dry transfer. Blot was blocked with 5% milk in TBS-T at 4°C/ON without agitation. Blot was incubated in the primary antibody at a dilution of 1:1000 in 5% TBS-T, at 4°C, ON without agitation. The antibody solution was decanted and the blot was rinsed briefly, then washed three times for 10 min in TBS-T at RT with agitation. Blot was incubated in Agrisera matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, AS09 602) diluted to 1:25 000 in TBS-T for 1h at RT with agitation. The blot was washed as above and developed for 2 min with Agrisera ECLBright. Exposure time was 92 seconds.
Courtesy of Eli Gordon, Elizabeth Vierling Lab Department of Biochemistry and Molecular Biology University of Massachusetts Amherst, USA

Additional information

Related products

Related products anti-tag antibodies

Background

Background Tagging protein with TurboID  allows to study protein interactions in different types of cells and organs and devvelopmental stages. This is a suitable tool for proximity labelling experiments as described in Mair et al. (2019). Proximity labeling of protein complexes and cell-type-specific organellar proteomes in Arabidopsis enabled by TurboID. Elife . 2019 Sep 19;8:e47864. doi: 10.7554/eLife.47864.

Product citations

Selected references To be added when available, antibody available in January 2021.

Related products: BirA (mutated/TurboID)

AS16 ECL-S-N | low pico to mid femtogram and extreme low femtogram detection
From 25 €
AS16 ECL-N |  low pico to mid femtogram detection
- 10ml trial size is restricted to on...
From 12 €
AS09 602 |  Clonality: Polyclonal | Host: Goat | Reactivity: Rabbit IgG (H&L)
194 €
AS17 4127 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Cas9/HA/LUC/C-YFP/N-YFP
607 €