Molecular mechanism of Afadin substrate recruitment to the receptor phosphatase PTPRK via its pseudophosphatase domain
- Cambridge Institute for Medical Research, University of Cambridge, United Kingdom
- Signalling Programme, Babraham Institute, United Kingdom
- Department of Pathology, University of Cambridge, United Kingdom
Figures
Figure 1 with 3 supplements
PTPRK interacts directly with the Afadin C-terminal coiled-coil (CC) region.
(A) Top: schematic of full-length human Afadin, with domain annotation based on UniProt ID P55196, showing Ras-association (RAS, yellow), CC (green), forkhead-associated (FHA, orange), dilute (DIL, …
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Figure 1—source data 1
Uncropped, unedited blots for Figure 1B.
- https://cdn.elifesciences.org/articles/79855/elife-79855-fig1-data1-v2.zip
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Figure 1—source data 2
Uncropped, unedited blots for Figure 1C.
- https://cdn.elifesciences.org/articles/79855/elife-79855-fig1-data2-v2.zip
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Figure 1—source data 3
Uncropped, unedited blots for Figure 1D.
- https://cdn.elifesciences.org/articles/79855/elife-79855-fig1-data3-v2.zip
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Figure 1—source data 4
Uncropped, unedited gels for Figure 1E.
- https://cdn.elifesciences.org/articles/79855/elife-79855-fig1-data4-v2.zip
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Figure 1—source data 5
Uncropped, unedited gels for Figure 1F.
- https://cdn.elifesciences.org/articles/79855/elife-79855-fig1-data5-v2.zip
Figure 1—figure supplement 1
Disorder predictions for Afadin.
Top: schematic of full-length human Afadin (also known as AF6 or MLLT4), with domain annotation based on UniProt ID P55196, colored as in main Figure 1A. The predicted local distance difference test …
Figure 1—figure supplement 2
Afadin-pY1230 antibody validation.
(A) Top: phosphopeptide antigen used to generate a sheep polyclonal anti-Afadin-pY1230 antibody (tthis study). Middle: amino acid numbers of epitope used to generate a mouse monoclonal against …
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Figure 1—figure supplement 2—source data 1
Uncropped, unedited blots for Figure 1—figure supplement 2B.
- https://cdn.elifesciences.org/articles/79855/elife-79855-fig1-figsupp2-data1-v2.zip
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Figure 1—figure supplement 2—source data 2
Uncropped, unedited blots for Figure 1—figure supplement 2C.
- https://cdn.elifesciences.org/articles/79855/elife-79855-fig1-figsupp2-data2-v2.zip
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Figure 1—figure supplement 2—source data 3
Uncropped, unedited blots for Figure 1—figure supplement 2D.
- https://cdn.elifesciences.org/articles/79855/elife-79855-fig1-figsupp2-data3-v2.zip
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Figure 1—figure supplement 2—source data 4
Densitometric quantification shown in Figure 1—figure supplement 2E.
- https://cdn.elifesciences.org/articles/79855/elife-79855-fig1-figsupp2-data4-v2.xlsx
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Figure 1—figure supplement 2—source data 5
Uncropped, unedited blots for Figure 1—figure supplement 2F.
- https://cdn.elifesciences.org/articles/79855/elife-79855-fig1-figsupp2-data5-v2.zip
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Figure 1—figure supplement 2—source data 6
Densitometric quantification shown in Figure 1—figure supplement 2G.
- https://cdn.elifesciences.org/articles/79855/elife-79855-fig1-figsupp2-data6-v2.xlsx
Figure 1—figure supplement 3
Purification of in vivo biotinylated PTP domains.
(A) Left: following Ni-NTA affinity chromatography, in vivo biotinylated PTPRK D1 and D2 domains were purified by size-exclusion chromatography (SEC) on an S75 16/600 column. Right: after SEC …
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Figure 1—figure supplement 3—source data 1
Uncropped, unedited gels for Figure 1—figure supplement 3A.
- https://cdn.elifesciences.org/articles/79855/elife-79855-fig1-figsupp3-data1-v2.zip
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Figure 1—figure supplement 3—source data 2
Uncropped, unedited blots for Figure 1—figure supplement 3B.
- https://cdn.elifesciences.org/articles/79855/elife-79855-fig1-figsupp3-data2-v2.zip
Figure 2 with 1 supplement
PTPRK:Afadin-CC forms an equimolar complex with micromolar affinity.
(A) Size-exclusion chromatography coupled to multi-angle light scattering (SEC-MALS) analysis of Afadin-CC. The SEC elution profile (normalized differential refractive index [dRI]; thin red line) …
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Figure 2—source data 1
Uncropped, unedited gels for Figure 2C.
- https://cdn.elifesciences.org/articles/79855/elife-79855-fig2-data1-v2.zip
Figure 2—figure supplement 1
GST-tag removal from Afadin-CC for size-exclusion chromatography coupled to multi-angle light scattering (SEC-MALS) and isothermal titration calorimetry (ITC) experiments.
(A) GST was removed from GST-Afadin-CC by overnight cleavage with GST-3C protease. Cleaved GST and GST-3C were removed by incubation with GSH-sepharose 4B (GSH-resin). (B) Residual cleaved GST was …
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Figure 2—figure supplement 1—source data 1
Uncropped, unedited gels for Figure 2—figure supplement 1A.
- https://cdn.elifesciences.org/articles/79855/elife-79855-fig2-figsupp1-data1-v2.zip
Figure 3 with 3 supplements
Structural prediction of the PTPRK-D2:Afadin-CC complex.
(A) Top: schematic of different GST-Afadin-CC truncations used for further interaction mapping experiments. Regions within the predicted helix are marked by a green bar. Dashed lines indicate …
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Figure 3—source data 1
Uncropped, unedited gels for Figure 3A.
- https://cdn.elifesciences.org/articles/79855/elife-79855-fig3-data1-v2.zip
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Figure 3—source data 2
Uncropped, unedited gel images for Figure 3D.
- https://cdn.elifesciences.org/articles/79855/elife-79855-fig3-data2-v2.zip
Figure 3—figure supplement 1
Structural prediction of the PTPRK-D2:Afadin-CC complex.
(A) Ribbon diagram of the AF2-Multimer prediction for the PTPRK-D2 domain (blue) in complex with the full Afadin-CC (aa. 1393–1455, green). Afadin-CC residues that contribute to the interaction …
Figure 3—figure supplement 2
Species conservation of Afadin-CC.
Multiple-sequence alignment of the equivalent Afadin-CC region (hsAFDN aa. 1393–1455) from human (Homo sapiens), mouse (Mus musculus), dog (Canis familiaris), frog (Xenopus laevis), zebrafish (Danio …
Figure 3—figure supplement 3
Prediction quality analysis of AF2 multimer-generated PTPRK-D2:Afadin-CC complex models.
(A) The five AF2-Multimer-generated PTPRK-D2:Afadin-CC models were superposed using the PTPRK-D2 chain only. The Afadin-CC chains of each ranked model are shown as cylinders (colored as indicated) …
Figure 4 with 4 supplements
PTPRK binds Afadin-CC via an acidic pocket distal from the D2 ‘active’ site.
(A) Conservation mapping of the PTPRK-D2:Afadin-CC interface. PTPRK-D2 is shown (blue surface representation) in complex with Afadin-CC (green ribbons). Residues that are conserved in both …
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Figure 4—source data 1
Uncropped, unedited gel for Figure 4D.
- https://cdn.elifesciences.org/articles/79855/elife-79855-fig4-data1-v2.zip
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Figure 4—source data 2
Uncropped, unedited gel for Figure 4E.
- https://cdn.elifesciences.org/articles/79855/elife-79855-fig4-data2-v2.zip
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Figure 4—source data 3
Uncropped, unedited blot for Figure 4G.
- https://cdn.elifesciences.org/articles/79855/elife-79855-fig4-data3-v2.zip
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Figure 4—source data 4
Uncropped, unedited gel images for Figure 4H.
- https://cdn.elifesciences.org/articles/79855/elife-79855-fig4-data4-v2.zip
Figure 4—figure supplement 1
Mapping of unique PTPRM residues.
Multiple-sequence alignment of human PTPRK, PTPRU, and PTPRM D2 domain sequences. Residues that are conserved in both PTPRK and PTPRU, but differ in PTPRM, which were used for interaction site …
Figure 4—figure supplement 2
Localization of additional PTPRK D2 mutations.
(A) Ribbon diagram of PTPRK-D2 (blue) and PTPRM-D2 (wheat) in complex with Afadin-CC (green). PTPRK-F1225 stacks with hydrophobic W1418-Y1419 residues of Afadin-CC and this residue is conserved in …
Figure 4—figure supplement 3
Thermal stability of PTPRK-D2 mutants.
Differential scanning fluorimetry of PTPRK-D2 WT (black), F1255A (red), M-loop (blue), and double mutant (DM, yellow) recombinant proteins. Curves and melting temperature (Tm) values shown represent …
Figure 4—figure supplement 4
Prediction quality of models prior to biochemical mapping.
(A) The 5 AF2-Multimer generated PTPRK-ICD:Afadin-CC models were superposed using the PTPRK-ICD chain only. The Afadin-CC chains of each ranked model are shown as cylinders (colored as indicated). (B…
Figure 5 with 1 supplement
Mutation of residues at the interaction interface inhibit dephosphorylation of Afadin by PTPRK.
(A) Time course of pNPP dephosphorylation by WT (black) and double mutant (DM, red) PTPRK-ICD. Error bars represent ± SEM of n = 2 independent experiments. (B) Immunoblot analysis of …
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Figure 5—source data 1
Uncropped, unedited blots for Figure 5B.
- https://cdn.elifesciences.org/articles/79855/elife-79855-fig5-data1-v2.zip
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Figure 5—source data 2
Uncropped, unedited blots for Figure 5C.
- https://cdn.elifesciences.org/articles/79855/elife-79855-fig5-data2-v2.zip
Figure 5—figure supplement 1
PTPRK-ICD-DM displays impaired dephosphorylation of Afadin-pY1230, but not p120Cat-pY228.
Densitometric quantification of immunoblots in Figure 5B (90 min time point shown) at indicated time points. Top: Afadin-pY1230 normalized against no protein control. Middle: p120Cat-pY228 …
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Figure 5—figure supplement 1—source data 1
Densitometric quantification of Figure 5—source data 1, shown in Figure 5—figure supplement 1.
- https://cdn.elifesciences.org/articles/79855/elife-79855-fig5-figsupp1-data1-v2.xlsx
Tables
Appendix 1—key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Gene (human) | PTPRK | ENSEMBL: ENSG00000152894 | ||
| Gene (human) | AFDN | ENSEMBL: ENSG00000130396 | ||
| Cell line (human) | MCF10A | ATCC | CRL-10317 | |
| Cell line (human) | HEK293T | D. Ron | N/A | |
| Transfected construct (human) | MCF10A PTPRK KO pooled.tGFP | Fearnley et al., 2019 | N/A | Lentivirally transduced stable cell line |
| Transfected construct (human) | MCF10A PTPRK KO pooled.tGFP. P2A.PTPRK | Fearnley et al., 2019 | N/A | Lentivirally transduced stable cell line |
| Transfected construct (human) | MCF10A PTPRK KO pooled.tGFP.P2A. PTPRK.C1089S | Fearnley et al., 2019 | N/A | Lentivirally transduced stable cell line |
| Transfected construct (human) | MCF10A tGFP | Fearnley et al., 2019 | N/A | Lentivirally transduced stable cell line |
| Antibody | Anti-PTPRK (rabbit monoclonal) | Fearnley et al., 2019 | 2.H4 | Western blot: 1:1000 |
| Antibody | Anti-Afadin-pY1230 (sheep polyclonal) | This study | N/A | Characterized in Figure 1—figure supplement 2 Western blot: 1 µg/ml (with 10 µg/ml non-phosphopeptide) Available on request from Sharpe lab, Babraham Institute |
| Antibody | Anti-Afadin (mouse monoclonal) | BD Transduction Laboratories | Cat#610732 | Western blot: 1:1000 |
| Antibody | Anti-p120 catenin (mouse monoclonal) | BD Transduction Laboratories | Cat#610133 | Western blot: 1:1000 |
| Antibody | Anti-RFP (mouse monoclonal) | Thermo Fisher Scientific | Cat#MA5-15257 | Western blot: 1:1000 |
| Antibody | Anti-Turbo-GFP (mouse monoclonal) | OriGene | TA150041 | Western blot: 1:1000 |
| Antibody | Anti-Afadin (rabbit monoclonal) | Cell Signaling Technology | Cat#13531 | Western blot: 1:1000 |
| Antibody | Anti-His (mouse monoclonal) | Cell Signaling Technology | Cat#2366 | Western blot: 1:1000 |
| Antibody | Anti-phospho-tyrosine (P-Tyr-1000) (rabbit monoclonal) | Cell Signaling Technology | Cat#8954 | Western blot: 1:2000 |
| Antibody | Anti-paxillin (rabbit monoclonal) | Cell Signaling Technology | Cat#12065 (D9G12) | Western blot: 1:1000 |
| Antibody | Anti-phospho-p120 catenin (Y904) (rabbit polyclonal) | Cell Signaling Technology | Cat#2910 | Western blot: 1:1000 |
| Antibody | Anti-phospho-p120 catenin (Y228) (rabbit polyclonal) | Cell Signaling Technology | Cat#2911 | Western blot: 1:1000 |
| Antibody | Anti-phospho- paxillin (Y118) | Cell Signaling Technology | Cat#2541 | Western blot: 1:1000 |
| Antibody | Anti-Tubulin (alpha) (mouse monoclonal) | Sigma | Cat#T6199 | Western blot: 1:1000 |
| Antibody | HRP-conjugated- donkey anti-goat IgG | Jackson ImmunoResearch | Cat#705-035-147 | Western blot: 1:5000 |
| Antibody | HRP-conjugated- donkey anti-rabbit IgG | Jackson ImmunoResearch | Cat#711-035-152 | Western blot: 1:5000 |
| Antibody | HRP-conjugated- donkey anti-mouse IgG | Jackson ImmunoResearch | Cat#711-035-152 | Western blot: 1:5000 |
| Antibody | HRP-conjugated-mouse anti- rabbit IgG (conformation specific) | Cell Signaling Technology | Cat#5127S | Western blot: 1:2000 |
| Recombinant DNA reagent | pET15b | J. Deane | N/A | |
| Recombinant DNA reagent | pET15b.His.TEV.Avi | Fearnley et al., 2019 | N/A | |
| Recombinant DNA reagent | pET15b.His.TEV.Avi. PTPRK.ICD | Fearnley et al., 2019 | UniProt: Q15262-3 | |
| Recombinant DNA reagent | pET15b.His.TEV.Avi. PTPRK.ICD.D1057A | Fearnley et al., 2019 | UniProt: Q15262-3 | |
| Recombinant DNA reagent | pET15b.His.TEV.Avi. PTPRK.ICD.C1089S | Fearnley et al., 2019 | UniProt: Q15262-3 | |
| Recombinant DNA reagent | pET15b.His.TEV.Avi. PTPRK.D1 | Fearnley et al., 2019 | UniProt: Q15262-3 | |
| Recombinant DNA reagent | pET15b.His.TEV.Avi. PTPRK.D2 | Fearnley et al., 2019 | UniProt: Q15262-3 | |
| Recombinant DNA reagent | PET15b.His.TEV.Avi. PTPRK.D2.G1273H | This study | UniProt: Q15262-3 | Mutations: G1273H See Figure 4 |
| Recombinant DNA reagent | pET15b.His.TEV.Avi. PTPRK.D2.L1335R | This study | UniProt: Q15262-3 | Mutations: L1335R See Figure 4D |
| Recombinant DNA reagent | pET15b.His.TEV.Avi. PTPRK.ICD.DM | This study | UniProt: Q15262-3 | Mutations: G1273H L1335R See Figure 4E |
| Recombinant DNA reagent | pET15b.His.TEV.Avi. PTPRK.D2.DM | This study | UniProt: Q15262-3 | Mutations: G1273H L1335R See Figure 4E |
| Recombinant DNA reagent | pET15b.His.TEV.Avi. PTPRK.D2.F1225A | This study | UniProt: Q15262-3 | Mutation: F1225A See Figure 4H |
| Recombinant DNA reagent | pET15b.His.TEV.Avi. PTPRK.D2.M-loop | This study | UniProt: Q15262-3 | Mutations: C1372Y E1373N E1374G See Figure 4H |
| Recombinant DNA reagent | pGEX-6P-1 | J. Deane | N/A | |
| Recombinant DNA reagent | pGEX-6P-Afadin-1098-C* | This study | UniProt: P55196-4 | See Figure 1E |
| Recombinant DNA reagent | pGEX-6P-Afadin-1098-1507 | This study | UniProt: P55196-4 | See Figure 1E |
| Recombinant DNA reagent | pGEX-6P-Afadin-1098-1407 | This study | UniProt: P55196-4 | See Figure 1 |
| Recombinant DNA reagent | pGEX-6P-Afadin-1514-C* | This study | UniProt: P55196-4 | See Figure 1E |
| Recombinant DNA reagent | pGEX-6P-Afadin-1393-C* | This study | UniProt: P55196-4 | See Figure 1E |
| Recombinant DNA reagent | pGEX-6P-Afadin-1393-1507 | This study | UniProt: P55196-4 | See Figure 1E |
| Recombinant DNA reagent | pGEX-6P-Afadin-CC | This study | UniProt: P55196-4 | Encoding amino acids: 1393–1455 See Figure 1F |
| Recombinant DNA reagent | pGEX-6P-Afadin-CC-WY>AA | This study | UniProt: P55196-4 | Mutations: W1418A Y1419A See Figure 3D |
| Recombinant DNA reagent | pGEX-6P-Afadin-CC-ER>AA | This study | UniProt: P55196-4 | Mutations: E1429A R1430A See Figure 3D |
| Recombinant DNA reagent | pGEX-6P-Afadin-CC-RK>AA | This study | UniProt: P55196-4 | Mutations: R1432A K1433A See Figure 3D |
| Recombinant DNA reagent | pGEX-6P-Afadin-CC-Q>A | This study | UniProt: P55196-4 | Mutations: Q1443A See Figure 3D |
| Recombinant DNA reagent | pGEX-6P-Afadin-CC-T>A | This study | UniProt: P55196-4 | Mutations: T1446A See Figure 3D |
| Recombinant DNA reagent | pGEX-6P-Afadin-CC-QT>AA | This study | UniProt: P55196-4 | Mutations: Q1443A T1446A See Figure 3D |
| Recombinant DNA reagent | pmScarlet-C1 | Z. Kadlecova | N/A | |
| Recombinant DNA reagent | pmScarlet-Afadin | This study | UniProt: P55196-4 | See Figure 1—figure supplement 2D |
| Recombinant DNA reagent | pmScarlet-Afadin-Y1226F | This study | UniProt: P55196-4 | Mutation: Y1226F See Figure 1—figure supplement 2D |
| Recombinant DNA reagent | pmScarlet-Afadin-Y1230F | This study | UniProt: P55196-4 | Mutation: Y1230F See Figure 1—figure supplement 2D |
| Recombinant DNA reagent | pmScarlet-Afadin-YF YF | This study | UniProt: P55196-4 | Mutations: Y1226F Y1230F See Figure 1—figure supplement 2D |
| Sequence-based reagent | ON-TARGETplus Human AFDN siRNA: | Dharmacon, GE Healthcare | L-020075-02-0005 | |
| Sequence-based reagent | ON-TARGETplus Non-targeting pool siRNA: | Dharmacon, GE Healthcare | Cat#D-001810-10-05 | |
| Peptide, recombinant protein | Catalase | Sigma | Cat#C134514 | |
| Peptide, recombinant protein | Cholera toxin | Sigma | Cat#C-8052 | |
| Peptide, recombinant protein | Insulin | Sigma | Cat#I-1882 | |
| Peptide, recombinant protein | Epidermal growth factor | PeproTech | Cat#AF-100-15-1MG | |
| Commercial assay or kit | Q5 High-Fidelity DNA Polymerase | New England Biolabs | Cat#M0491S | |
| Commercial assay or kit | Phusion Hot Start II DNA polymerase | Thermo Fisher Scientific | Cat#F549L | |
| Commercial assay or kit | EZ-ECL substrate | Geneflow | Cat#K1-0170 | |
| Commercial assay or kit | InstantBlue | Expedeon | Cat#ISB1L | |
| Commercial assay or kit | Phosphatase inhibitor cocktail | Roche | Cat#04906845001 | |
| Commercial assay or kit | TaqMan Universal Master Mix II | Applied Biosystems | Cat#4440040 | |
| Chemical compound, drug | Hydrogen peroxide | Thermo Fisher Scientific | Cat#H/1750/15 | |
| Chemical compound, drug | Sodium orthovanadate | Alfa Aesar | Cat#J60191 | |
| Chemical compound, drug | 250 kDa-FITC-dextran | Sigma | Cat#FD250S-100MG | |
| Chemical compound, drug | Para-Nitrophenol- phosphate (pNPP) | New England Biolabs | Cat#P0757 | |
| Chemical compound, drug | IPTG | Generon | Cat#GEN-S-02122 | |
| Chemical compound, drug | D-biotin | Sigma | Cat#B4639 | |
| Chemical compound, drug | l-Glutamine | Sigma | Cat#G7513 | |
| Chemical compound, drug | Hydrocortisone | Sigma | Cat#H-0888 | |
| Chemical compound, drug | NH4OH | Acros Organics | Cat#460801000 | |
| Chemical compound, drug | Methanol-free 16% (w/v) paraformaldehyde (PFA) | Thermo Fisher Scientific | Cat#28906 | |
| Software, algorithm | FIJI/ImageJ | Laboratory for Optical and Computational Instrumentation | University of Wisconsin-Madison | |
| Software, algorithm | GraphPad | Prism | ||
| Software, algorithm | Chimera | UCSF | ||
| Strain, strain background (Escherichia coli) | STABLE competent E. coli | NEB | Cat#C3040I | |
| Strain, strain background (E. coli) | DH5alpha competent E. coli | Invitrogen | Cat#18265017 | |
| Strain, strain background (E. coli) | BL21 DE3 Rosetta E. coli | J. Deane | N/A | |
| Other | DMEM | Thermo Fisher Scientific | Cat#41965-039 | Component of cell culture media |
| Other | Ham's F-12 | Sigma | Cat#N4888 | Component of cell culture media |
| Other | Horse serum | Thermo Fisher Scientific | Cat#16050-122 | Component of cell culture media |
| Other | HRP-conjugated streptavidin | Thermo Fisher Scientific | Cat#434323 | For detection of biotinylated proteins |
| Other | Fetal bovine serum | Sigma | Cat#F7524-500ml | Component of cell culture media |
| Other | Trypsin-EDTA solution | Sigma | Cat#T3924 | Reagent used to lift cells from culture vessel |
| Other | GeneJuice | Merck Millipore | Cat#70967-3 | Transfection reagent |
| Other | EDTA-free protease inhibitors | Roche | Cat#11836170001 | Component of cell lysis buffer |
| Other | Lipofectamine RNAiMAX | Invitrogen | Cat#13778075 | Transfection reagent |
| Other | OptiMEM | Thermo Fisher Scientific | Cat#31985070 | Component of cell culture media |
| Other | Protein G agarose beads | Merck Millipore | Cat#16-266 | Affinity reagent for immunoprecipitations |
| Other | Ni-NTA agarose | QIAGEN | Cat#1018244 | Affinity reagent for His-tag purification |
| Other | Streptavidin-coated magnetic beads | New England Biolabs | Cat#S1420S | Affinity reagent for Avi-tag pull-downs |
| Other | Streptavidin agarose | Thermo Fisher Scientific | Cat#20357 | Affinity reagent for Avi-tag pull-downs |
| Other | Superdex 200 16/600 column | GE Healthcare | Cat#28-9893-35 | Chromatography column |
| Other | Superdex 75 16/600 column | GE Healthcare | Cat#28-9893-33 | Chromatography column |
| Other | Ultracel-3K regenerated cellulose centrifugal filter | Merck Millipore | Cat#UFC900324 | Chromatography column |
| Other | Ultracel-10K regenerated cellulose centrifugal filter | Merck Millipore | Cat#UFC901024 | Used for protein concentration |
| Other | Ultracel-30K regenerated cellulose centrifugal filter | Merck Millipore | Cat#UFC903024 | Used for protein concentration |
| Other | NuPAGE 4–12% Bis-Tris gel | Thermo Fisher Scientific | Cat#NP0321BOX | SDS PAGE electrophoresis gel |
| Other | Slide-A-Lyzer 20K MWCO | Thermo Fisher Scientific | Cat#66003 | Dialysis cassette |
| Other | SYPRO Orange dye | Thermo Fisher Scientific | Cat#S6650 | Used for thermal shift assays |
| Other | MycoAlert PLUS Mycoplasma Detection Kit | Lonza | #LT07-705 | Used for testing cell lines |
| Other | MycoProbe Mycoplasma Detection Kit | R&D Systems | #CUL001B | Used for testing cell lines |
