- PA5-20344 - Invitrogen Antibodies PDCD1LG2 antibody

Jang HR, Shin SB, Kim CH, Won JY, Xu R, Kim DE, Yim H
Cell death and differentiation 2021 Sep;28(9):2745-2764

Wu J, Sun L, Li H, Shen H, Zhai W, Yu Z, Chen G
Journal of neuroinflammation 2017 Feb 14;14(1):36

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of Raji cell lysate using a CD273/B7 DC polyclonal antibody (Product # PA5-20344) at (A) 0.5, (B) 1 and (C) 2 µg/mL.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of CD273 in HeLa WT or PD-L2 KO cells. Lysates (15 µg) were loaded onto SDS-PAGE and blots were probed with CD273 (B7-DC) Polyclonal Antibody (Product # PA5-20344) diluted to 4 µg/mL and anti-beta actin diluted to 1 µg/mL. 1 h incubation at RT in 0.05 NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10,000 dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of CD273 in HeLa WT or PD-L2 KO cells. Lysates (15 µg) were loaded onto SDS-PAGE and blots were probed with CD273 (B7-DC) Polyclonal Antibody (Product # PA5-20344) diluted to 4 µg/mL and anti-beta actin diluted to 1 µg/mL. 1 h incubation at RT in 0.05 NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10,000 dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot Validation in Human Raji Cell Lysate. Loading: 15 µg of lysates per lane. Antibodies: CD273 (B7-DC) Polyclonal Antibody (Product # PA5-20344) (A: 0.5 µg/mL and B: 1 µg/mL), 1h incubation at RT in 0.05 NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10,000 dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of mouse brain cells using a CD273/B7 DC polyclonal antibody (Product # PA5-20344) at a 20 µg/mL dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of CD273 (B7DC) was performed using 70% confluent log phase Jurkat cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with CD273 (B7DC) Monoclonal Antibody (176611) (Product # PA5-20344) at 20 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415). Panel d represents the merged image showing membranous localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of CD273 (B7-DC) was performed using formalin-fixed paraffin-embedded human colon (diffuse large B cell lymphoma) tissue sections. To expose the target protein, heat-induced epitope retrieval was performed on de-paraffinized sections using eBioscience™ IHC Antigen Retrieval Solution - Low pH (10X) (Product # 00-4955-58) diluted to 1X solution in water in a decloaking chamber at 110 degree Celsius for 15 minutes. Following antigen retrieval, the sections were blocked with 2% normal goat serum in 1X PBS for 45 minutes at room temperature and then probed with or without CD273 (B7-DC) Polyclonal Antibody (Product # PA5-20344) at 10 µg/mL concentration in 0.1% normal goat serum overnight at 4 degree Celsius in a humidified chamber. Detection was performed using Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731) at a dilution of 1:2000 in 0.1% normal goat serum for 45 minutes at room temperature. ReadyProbes™ Tissue Autofluorescence Quenching Kit (Product # R37630) was used to quench autofluorescence from the tissues. Nuclei were stained with DAPI (Product # D1306) and the sections were mounted using ProLong™ Glass Antifade Mountant (Product # P36984). The images were captured on EVOS™ M7000 Imaging System (Product # AMF7000) at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse brain cells labeling PD-L2 with CD273 (B7-DC) Polyclonal Antibody (Product # PA5-20344) at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1:500 dilution (red).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse brain tissue labeling PD-L2 with CD273 (B7-DC) Polyclonal Antibody (Product # PA5-20344) at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1:500 dilution (green) and DAPI staining (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of paraffin-embedded mouse brain tissue using CD273 (B7-DC) Polyclonal Antibody (Product # PA5-20344) at 2.5 µg/mL. Tissue was fixed with formaldehyde and blocked with 0.1 serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of paraffin-embedded mouse brain tissue using CD273 (B7-DC) Polyclonal Antibody (Product # PA5-20344) at 2.5 µg/mL. Tissue was fixed with formaldehyde and blocked with 0.1 serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 2 ICH increased the protein levels of PD-1/PD-Ls and the interaction between PD-1 and PD-L1. a Time course of the protein levels of PD-1, PD-L1, and PD-L2 in the brain tissue around hematoma after ICH. Representative western blot bands of PD-1, PD-L1, and PD-L2 and quantitative analysis of the relative protein level were shown. The mean value of sham group was normalized to 1.0. Data are expressed as mean +- SEM, n = 6. Double asterisks indicate p < 0.01, triple asterisks indicate p < 0.001 vs. sham group. b , c Immunoprecipitation analysis of the interaction between PD-1 and PD-L1 at indicated times after ICH. All values are means +- SEM, n = 6. triple asterisks indicate p < 0.001 vs. sham group, triple pound signs indicate p < 0.001

PA5-20344