Nature - 15.08.2019

3

nature research | reporting summary

October 2018

Ubiquitin Ubi-1 from abcam (Cat:ab7254), validation statement from the manufacturer:

Tested applications Suitable for: IHC-P, WB, ELISA, ICC/IF, IHC-Fr

Unsuitable for: IP

Species reactivity Reacts with: Mouse, Rat, Chicken, Cow, Human, Arabidopsis thaliana, Caenorhabditis elegans, Drosophila

melanogaster, Drosophila C virus

validation found at provider's website: https://www.abcam.com/ubiquitin-antibody-ubi-1-ab7254.html

CS (cell signaling technology)-Ub and abcam-Ub antibodies were validated in Bhogaraju et al., 2106 Cell for the purpose of

monitoring SdeA-mediated ubiquitin modification in HEK293T cells.

GFP (B-2) Santa Cruz Biotechnology (sc-9996)

Validation statement from the manufacturer: recommended for detection of GFP and GFP mutant fusion proteins by WB, IP, IF,

FCM and ELISA. Validation found at provider's website https://www.scbt.com/scbt/product/gfp-antibody-b-2?

productCanUrl=gfp-antibody-b-2&_requestid=3395142

Anti-tubulin

Validation statement from the manufacturer: β-Tubulin (9F3) Rabbit mAb detects endogenous levels of total β-tubulin protein,

and does not cross-react with recombinant α-tubulin.

Species Reactivity:

Human, Mouse, Rat, Monkey, Zebrafish, Bovine Validation found at provider's website: https://www.cellsignal.com/products/

primary-antibodies/b-tubulin-9f3-rabbit-mab/2128

Anti-HA

Validation statement from the manufacturer: specific to epitope mapping within an internal region of the the influenza

hemagglutinin (HA) protein protein, recommended for detection of proteins containing the HA tag by WB, IP, IF, FCM and ELISA

Validation found at provider's website: https://www.scbt.com/scbt/product/ha-probe-antibody-f-7?productCanUrl=ha-probe-

antibody-f-7&_requestid=3428134

Anti-CaM

Validation statement from the manufacturer: Calmodulin Antibody detects endogenous levels of total calmodulin protein.

Species Reactivity:

Mouse, Rat

Species predicted to react based on 100% sequence homology:

Human, Monkey, Xenopus, Pig

Validation found at provider's website: https://www.cellsignal.com/products/primary-antibodies/calmodulin-antibody/4830

Anti-GST

Validation statemet from the manufacturer: GST Antibody detects transfected GST fusion proteins.

Validation found at provider's website: https://www.cellsignal.com/products/primary-antibodies/gst-antibody/2622

GFP-Trap beads:

Validation statemet from the manufacturer : GFP-Trap® Agarose is an affinity resin for immunoprecipitation of GFP-fusion

proteins.

It consists of a GFP Nanobody/ VHH coupled to agarose beads.

Specificity

GFP, EGFP, CFP, YFP, BFP and many more derivatives

Validation found at provider's website: https://www.chromotek.com/products/detail/product-detail/gfp-trap-agarose/

Eukaryotic cell lines

Policy information about cell lines

Cell line source(s) RAW 264.7 cells (ATCC® TIB-71™), A549 cells (ATCC® CCL-185™) and HEK293T (ATCC® CRL-3216™).

Authentication Cell lines were authenticated using STR DNA profiling.

Mycoplasma contamination All the cell lines used tested negative for mycoplasma.

Commonly misidentified lines

(See ICLAC register)

The cell lines used in the study are not in the commonly misidentified lines list.