Indirect ELISA

Figure 2. Western blot analysis od Dityrosine -cross-linked Human Amyloid Beta. 200 ng of Abeta 1-40 peptides before and after Cu (II)-Oxidation were separated by SDS-PAGE and immunblotted with AM20243PU-S. AM20243PU-S detects dimeric and trimeric Abeta 1-4

Western blot with Porcine plasma and purified Porcine CRP (poCRP). Porcine plasma was diluted 1/25 and spiked with 0.08 or 0.3ug purified Porcine CRP, respectively. Polyacrylamide gel electrophoresis was performed under reducing and denaturing conditions on a 10-20% gel (Novex), blotted onto nitrocellulose and developed with monoclonal antibody PC1 (left) or competitor's monoclonal antibody (right). Lane A: Porcine plasma spiked with 0.08ug poCRP; lane B: 0.08ug poCRP; lane C: Porcine plasma spiked with 0.3ug poCRP; lane D: Porcine plasma; lane E: 0.3ug poCRP. Monoclonal antibody PC1 barely reacts with with the 50-55kDa bands visible on the right, presumably immunoglobulin heavy chains. Gel was slightly overloaded to allow visibility of CRP band in plasma.

Immunohistochemistry: AM20226PU 8-OHdG antibody staining of Paraffin Section of UV-irradiated Mouse Skin using an antigen retrieval in 0.1% Trypsin. The section was incubated with AM20226PU (5 g/ml), followed by staining with Avidin-Biotin-Complex method.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY CD274 (RC213071, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-CD274 (1:500). Positive lysates LY415473 (100ug) and LC415473 (20ug) can be purchased separately from OriGene.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY CD274 (RC213071, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-CD274 (1:500). Positive lysates LY415473 (100ug) and LC415473 (20ug) can be purchased separately from OriGene.

HEK293T cells were transfected with the pCMV6-ENTRY control (Cat# PS100001, Left lane) or pCMV6-ENTRY CD274 (Cat# RC213071, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-CD274(Cat# TA507087). Positive lysates LY415473 (100ug) and LC415473 (20ug) can be purchased separately from OriGene.

HEK293T cells were transfected with the pCMV6-ENTRY control (Cat# PS100001, Left lane) or pCMV6-ENTRY CD274 (Cat# RC213071, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-CD274(Cat# TA507087). Positive lysates LY415473 (100ug) and LC415473 (20ug) can be purchased separately from OriGene.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY PDCD1 (RC210364, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-PDCD1 (1:2000). Positive lysates LY401555 (100ug) and LC401555 (20ug) can be purchased separately from OriGene.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY PON1 (RC210356, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-PON1 (TA502528). Positive lysates LY400156 (100ug) and LC400156 (20ug) can be purchased separately from OriGene.

PD1 ELISA with 1B11 Capture (CF807867) and 21F5 Detection (CF806927) Antibodies. Substrate used: Recombinant Human PD1 (TP310364)

CD80 ELISA with 7D6 (TA812242) Capture and 11G5 (TA812246) Detection Antibodies. Substrate used: Recombinant Human CD80 (TP700242)

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY PDCD1 (RC210364, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-PDCD1 (1:2000). Positive lysates LY401555 (100ug) and LC401555 (20ug) can be purchased separately from OriGene.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY PDCD1 (RC210364, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-PDCD1. Positive lysates LY401555 (100ug) and LC401555 (20ug) can be purchased separately from OriGene.

Figure A, Western blot analysis of overexpressed lysates (15ug per lane) from HEK293T cells transfected with empty plasmid (PS100001, NC), human FOLH1 plasmid (RC218310, hFOLH1), mouse FOLH1 plasmid (MR216018, mFOLH1) using anti-FOLH1 antibody TA504509 (1:10000@1mg/ml). Figure B, Western blot analysis of the same samples as figure A with anti-DDK antibody (TA180144, 1:10000@1mg/ml)

PD1 ELISA with 1B11 Capture (CF807867) and 21F5 Detection (CF806927) Antibodies. Substrate used: Recombinant Human PD1 (TP310364)

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY GFAP (RC204548, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-GFAP. Positive lysates LY419563 (100ug) and LC419563 (20ug) can be purchased separately from OriGene.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY B2M (RC207587, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-B2M. Positive lysates LY401311 (100ug) and LC401311 (20ug) can be purchased separately from OriGene.

Immunocytochemistry staining of CD274 stable expression cells using anti-CD274 mouse monoclonal antibody (TA507087) (Left). The right is negative control (1:5000).

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY CD274 (RC213071, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-CD274 (1:500). Positive lysates LY415473 (100ug) and LC415473 (20ug) can be purchased separately from OriGene.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY PON1 (RC210356, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-PON1 (TA502528). Positive lysates LY400156 (100ug) and LC400156 (20ug) can be purchased separately from OriGene.

Immunocytochemistry staining of CD274 stable expression cells using anti-CD274 mouse monoclonal antibody (TA507087) (Left). The right is negative control (1:5000).

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY BRAF (RC211013, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-BRAF. Positive lysates LY401382 (100ug) and LC401382 (20ug) can be purchased separately from OriGene.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY PON1 (RC210356, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-PON1. Positive lysates LY400156 (100ug) and LC400156 (20ug) can be purchased separately from OriGene.

Western blot analysis of extracts from 293-6E cell line (left lane) and 293-6E cells transfected with UCHL1 gene (right lane), using anti- UCHL1 rabbit monoclonal antibody (TA591048) at 1:500 dilution.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY GFAP (RC204548, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-GFAP. Positive lysates LY419563 (100ug) and LC419563 (20ug) can be purchased separately from OriGene.

Western blot analysis of extracts from 293-6E cell line (left lane) and 293-6E cells transfected with WT1 gene (right lane), using anti- WT1 rabbit monoclonal antibody (TA591047) at 1:500 dilution.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY BRAF (RC211013, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-BRAF. Positive lysates LY401382 (100ug) and LC401382 (20ug) can be purchased separately from OriGene.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY CD80 (RC206540, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-CD80 (1:500).

Figure A, Western blot analysis of overexpressed lysates (15ug per lane) from HEK293T cells transfected with empty plasmid (PS100001, NC), human FOLH1 plasmid (RC218310, hFOLH1), mouse FOLH1 plasmid (MR216018, mFOLH1) using anti-FOLH1 antibody TA504509 (1:10000@1mg/ml). Figure B, Western blot analysis of the same samples as figure A with anti-DDK antibody (TA180144, 1:10000@1mg/ml)

SARS-Cov-2 RBD neutralizing antibody (Cat# TA814512) inhibits human ACE2 protein (Cat# TP701115) binding to SARS-CoV-2 RBD protein TP701119). The IC50 is typically 0.91 nM.

Western blot analysis of full length recombinant SARS-CoV-2 N protein(Cat# TP790189, 0.02ug) by using anti-SARS-CoV-2 N protein antibody (Cat# TA890226). (1:2000)

CD80 ELISA with 7D6 (TA812242) Capture and 11G5 (TA812246) Detection Antibodies. Substrate used: Recombinant Human CD80 (TP700242)

Western blot analysis of full length recombinant SARS-CoV-2 N protein(Cat# TP790189, 0.02ug) by using anti-SARS-CoV-2 N protein antibody (Cat# TA890226). (1:2000)

CD80 ELISA with 1A10 (TA812244) Capture and 11G5 (TA812246) Detection Antibodies. Substrate used: Recombinant Human CD80 (TP700242)

Western blot analysis of recombinant SARS-CoV-2 spike-RBD protein (Cat# TP750182, 0.02ug) by using anti-SARS-CoV-2 spike protein antibody (Cat# TA592329).

HEK293T cells were transfected with the pCMV6-ENTRY control (Cat# PS100001, Left lane) or pCMV6-ENTRY CD274 (Cat# RC213071, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-CD274(Cat# TA507087). Positive lysates LY415473 (100ug) and LC415473 (20ug) can be purchased separately from OriGene.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY PON1 (RC210356, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-PON1. Positive lysates LY400156 (100ug) and LC400156 (20ug) can be purchased separately from OriGene.

Western blot analysis is shown using Collagen type I antibody to detect expression of collagen I in Wistar rat hepatic stellate cells (HSC) in control (GFP-transduced) (left lane) and PPARgamma-transduced cell lysates (right lane). Protein staining shown below each blot depicts equal protein loading. An equal amount of the whole cell protein (100 ug) was separated by SDS-PAGE and electroblotted to nitrocellulose membranes. Proteins were detected by incubating the membrane with Collagen type I antibody at a concentration of 0.2-2 ug/10 ml in TBS (100 mM Tris-HCl, 0.15 M NaCl, pH 7.4) with 5% Non-fat milk. Detection occurred by incubation with a horseradish peroxidase-conjugated secondary antibody at 1 ug/10 ml. Proteins were detected by a chemiluminescent method using the PIERCE ECL kit (Amersham Biosciences). Other detection systems will yield similar results. See Hazra et al. (2004) for additional details.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY CD80 (RC206540, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-CD80 (1:500).

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY FH (RC200614, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-FH. Positive lysates LY400053 (100ug) and LC400053 (20ug) can be purchased separately from OriGene.

HEK293T cells were transfected with the pCMV6-ENTRY control (Cat# PS100001, Left lane) or pCMV6-ENTRY LIMK2 (Cat# RC217705, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-LIMK2(Cat# TA590208).

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY PROSC (RC200853, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-PROSC. Positive lysates LY416132 (100ug) and LC416132 (20ug) can be purchased separately from OriGene.

Standard curve for ELISA analysis with FOLH1 recombinant protein (dilution range from 5ng/ml to 5ug/ml) using FOLH1 Capture Antibody (Cat# TA504567) at 5ug/ml and HRP conjugated FOLH1 Detection mAb (Cat# TA504550) at 0.03ug/ml.

PD1 ELISA with 1B11 Capture (CF807867) and 21F5 Detection (CF806927) Antibodies. Substrate used: Recombinant Human PD1 (TP310364)

ELISA: HDAC2 Antibody

HEK293T cells were transfected with the pCMV6-ENTRY control (Cat# PS100001, Left lane) or pCMV6-ENTRY S100A1 (Cat# RC200278, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-S100A1 (Cat# TA801079). Positive lysates LY401900 (100ug) and LC401900 (20ug) can be purchased separately from OriGene.

CD80 ELISA with 7D6 (TA812242) Capture and 11G5 (TA812246) Detection Antibodies. Substrate used: Recombinant Human CD80 (TP700242)

Western blot analysis of recombinant SARS-CoV-2 Spike-RBD protein (Cat#TP750182, 0.02ug) by using anti-SARS-CoV-2 spike protein antibody (TA814463). (1：1000)

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY PDCD1 (RC210364, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-PDCD1 (1:2000). Positive lysates LY401555 (100ug) and LC401555 (20ug) can be purchased separately from OriGene.