Nature - USA (2020-01-16)

(Antfer) #1

Life Sciences Reporting Summary (Antibodies section did not have enough room)



  1. Antibodies


Describe the antibodies used and how they were validated


for use in the system under study (i.e. assay and species).


All antibodies used in this study are commercially available. Antibody validation was performed by the


individual manufacturer; validation information can be found on the manufacturers’ web pages via the


links provided below. Please also see Supplementary Table 1 and Online Methods for further description


of antibodies, including dilutions used. Staining of samples in this study was also compared to unstained


or secondary antibody-only controls processed in the same way.


Individual validation statements:


Abcam: https://www.abcam.com/primary-antibodies/improving-reproducibility-with-better-antibodies


Thermo Fisher: https://www.thermofisher.com/us/en/home/life-science/antibodies/invitrogen-


antibody-validation.html


Novus: https://www.novusbio.com/5-pillars-validation


BioLegend: https://www.biolegend.com/reproducibility


eBioscience: https://www.thermofisher.com/us/en/home/life-science/antibodies/invitrogen-antibody-


validation.html


Assay: Immunohistochemistry


CD68 - Figures 1b, 1c, 3i, Extended Data Figure 6b.


Supplier: Abcam, #ab53444


Monoclonal, clone FA-11


Lot: GR3241798-1


Manufacturer’s validation information: https://www.abcam.com/cd68-antibody-fa-11-ab53444.pdf


Dilution used here: 1:100


Detected with:


Goat anti-rat IgG Alexa Fluor 488 (Figures 1b, 1c)


Supplier: Life Technologies, #A-11006


Lot: 1887148


Manufacturer’s validation information: https://assets.thermofisher.com/TFS-


Assets/LSG/certificate/Certificates-of-Analysis/1887148_A11006.pdf


Dilution used here: 1:200


or


Goat anti-rat IgG Alexa Fluor 647 (Figure 3i, Extended Data Figure 6b)


Supplier: Life Technologies, #A-21247


Lot: 2043368


Manufacturer’s validation information: https://assets.thermofisher.com/TFS-


Assets/BID/certificate/Certificates-of-Analysis/A21247%20Lot%202043368%20CofA.pdf


Dilution used here: 1:200


Ki-67 - Figures 2b, 2c.

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