Western Blot (WB)
(Figure 1 Western Blot Validation of PD-L1 in HeLa CellsLoading: 15ug of lysates per lane. Antibodies: MBS151123 (1 ug/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.)
Rabbit PDL-1 Polyclonal Antibody | anti-CD274 antibody
PDL-1 Antibody
siRNA Knockdown Validation (Figure 3): Anti-PD-L1 antibody (MBS151123) specificity was further verified by PD-L1 specific siRNA knockdown. PD-L1 signal in HeLa cells transfected with PD-L1 siRNAs was much weaker in comparison with that in HeLa cells transfected with control siRNAs.
Overexpression Validation (Figure 4): Anti-PD-L1 antibodies (MBS151123) can detect the overexpression of PD-L1 protein in 293 cells transfected with PD-L1.
Western Blot (WB)
(Figure 1 Western Blot Validation of PD-L1 in HeLa CellsLoading: 15ug of lysates per lane. Antibodies: MBS151123 (1 ug/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.)
Western Blot (WB)
(Figure 1 Western Blot Validation of PD-L1 in HeLa CellsLoading: 15ug of lysates per lane. Antibodies: MBS151123 (1 ug/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.)
Western Blot (WB)
(Figure 2 Independent Antibody Validation (IAV) via Protein Expression Profile in Human and Mouse cell linesLoading: 15 ug of lysates per lane. Antibodies: MBS151123 (2ug/mL), MBS154572 (2 ug/mL), and beta-actin (1 ug/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit and or anti-mouse IgG HRP conjugate at 1:10000 and 1:5000 dilution, respectively.)
Western Blot (WB)
(Figure 2 Independent Antibody Validation (IAV) via Protein Expression Profile in Human and Mouse cell linesLoading: 15 ug of lysates per lane. Antibodies: MBS151123 (2ug/mL), MBS154572 (2 ug/mL), and beta-actin (1 ug/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit and or anti-mouse IgG HRP conjugate at 1:10000 and 1:5000 dilution, respectively.)
Western Blot (WB)
(Figure 3 Validation with PD-L1 siRNA Knockdown in HeLa CellsHeLa cells were transfected with control siRNAs (lane 1) orPD-L1 siRNAs (lane 2) Loading: 10 ug of HeLa whole cell lysates per lane. Antibodies: MBS154572 (2 ug/mL) and GAPDH (MBS150773, 0.02 ug/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-mouse IgG HRP conjugate at 1:5000 dilution.)
Western Blot (WB)
(Figure 3 Validation with PD-L1 siRNA Knockdown in HeLa CellsHeLa cells were transfected with control siRNAs (lane 1) orPD-L1 siRNAs (lane 2) Loading: 10 ug of HeLa whole cell lysates per lane. Antibodies: MBS154572 (2 ug/mL) and GAPDH (MBS150773, 0.02 ug/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-mouse IgG HRP conjugate at 1:5000 dilution.)
Western Blot (WB)
(Figure 4 Validation with PD-L1 over expression in 293 cellsLoading: Lysates/proteins at 15 μg per lane. Lane 1: non-transfected 293 cells Lane 2: PD-L1 overexpressed 293 cells Antibodies: MBS151123 (1 μg/mL). 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgGHRP conjugate at 1:10000 dilution.)
Western Blot (WB)
(Figure 4 Validation with PD-L1 over expression in 293 cellsLoading: Lysates/proteins at 15 μg per lane. Lane 1: non-transfected 293 cells Lane 2: PD-L1 overexpressed 293 cells Antibodies: MBS151123 (1 μg/mL). 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgGHRP conjugate at 1:10000 dilution.)
Immunohistochemistry (IHC)
(Figure 5 Immunohistochemistry Validation of PD-L1 in Human Tonsil CellsImmunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-PD-L1 antibody (MBS151123) at 5ug/ml. Tissue was fixed with formalde hyde and blocked with 10% 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.)
Immunohistochemistry (IHC)
(Figure 5 Immunohistochemistry Validation of PD-L1 in Human Tonsil CellsImmunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-PD-L1 antibody (MBS151123) at 5ug/ml. Tissue was fixed with formalde hyde and blocked with 10% 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.)
Immunofluorescence (IF)
(Figure 6 Immunofluorescence Validation of PD-L1 in Human HeartImmunofluorescent analysis of 4% paraformaldehyde-fixed human heart tissue labeling PD-L1 with MBS151123 at 20μg/mL, followed by goat anti-rabbit IgG secondaryantibody at 1/500 dilution (red). Image showing both membrane and cytoplasmic staining on human heart tissue.)
Immunofluorescence (IF)
(Figure 6 Immunofluorescence Validation of PD-L1 in Human HeartImmunofluorescent analysis of 4% paraformaldehyde-fixed human heart tissue labeling PD-L1 with MBS151123 at 20μg/mL, followed by goat anti-rabbit IgG secondaryantibody at 1/500 dilution (red). Image showing both membrane and cytoplasmic staining on human heart tissue.)
Flow Cytometry (FC)
(Figure 7 Flow Cytometry Validation of PD-L1Overlay histogram showing A-20 cells stained with MBS151123 (red line, 1ug/1x106 cells). 1 h incubation at 4°C in 2% FBS/PBS. Followed by secondary antibody 488 goat anti-rabbit IgG (H+L) at 1/500 dilution for 1 h 4°C. Isotype control antibody (Green line) was mouse IgG1 (1ug/1x106 cells) used under the same conditions. Acquisition of >10,000 events was performed.)
Flow Cytometry (FC)
(Figure 7 Flow Cytometry Validation of PD-L1Overlay histogram showing A-20 cells stained with MBS151123 (red line, 1ug/1x106 cells). 1 h incubation at 4°C in 2% FBS/PBS. Followed by secondary antibody 488 goat anti-rabbit IgG (H+L) at 1/500 dilution for 1 h 4°C. Isotype control antibody (Green line) was mouse IgG1 (1ug/1x106 cells) used under the same conditions. Acquisition of >10,000 events was performed.)
Immunohistochemisty (IHC)
(Figure 8 Immunohistochemistry Validation of PD-L1 in Rat HeartImmunohistochemical analysis of paraffin-embedded rat heart tissue using anti-PD-L1 antibody (MBS151123) at 5 ug/ml.Tissue was fixed with formaldehyde and blocked with 10% 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 goatanti-rabbit IgG H&L (HRP) at 1/250 was used as secondary.Counter stained with Hematoxylin)
Immunohistochemisty (IHC)
(Figure 8 Immunohistochemistry Validation of PD-L1 in Rat HeartImmunohistochemical analysis of paraffin-embedded rat heart tissue using anti-PD-L1 antibody (MBS151123) at 5 ug/ml.Tissue was fixed with formaldehyde and blocked with 10% 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 goatanti-rabbit IgG H&L (HRP) at 1/250 was used as secondary.Counter stained with Hematoxylin)
Immunohistochemistry (IHC)
(Figure 9 Immunohistochemistry Validation of PD-L1 in Human HeartImmunohistochemical analysis of paraffin-embedded human heart tissue using anti-PD-L1 antibody (MBS151123) at 2.5 °g/ml. Tissue was fixed with formalde hyde and blocked with 10% 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 assecondary. Counter stained with Hematoxylin.)
Immunohistochemistry (IHC)
(Figure 9 Immunohistochemistry Validation of PD-L1 in Human HeartImmunohistochemical analysis of paraffin-embedded human heart tissue using anti-PD-L1 antibody (MBS151123) at 2.5 °g/ml. Tissue was fixed with formalde hyde and blocked with 10% 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 assecondary. Counter stained with Hematoxylin.)
Immunofluorescence (IF)
(Figure 10 Immunofluorescence Validation of PD-L1 in Rat HeartImmunofluorescence analysis of 4% paraformaldehyde-fixed rat heart tissue labeling PD-L1 with MBS151123 at 20 ug/ml,followed by goat anti-rabbit IgG secondary antibody at 1/250 dilution (red).)
Immunofluorescence (IF)
(Figure 10 Immunofluorescence Validation of PD-L1 in Rat HeartImmunofluorescence analysis of 4% paraformaldehyde-fixed rat heart tissue labeling PD-L1 with MBS151123 at 20 ug/ml,followed by goat anti-rabbit IgG secondary antibody at 1/250 dilution (red).)
Immunofluorescence (IF)
(Figure 11 Immunofluorescence Validation of PD-L1 intumors in Human Cells (Dhar et al., 2018)(A) Several antibody brands were first tested with RBCs,WBCs, and HeLa cells. (MBS151123) was chosen as it provided the highest staining intensity. (B, C) Using the optimal conditions of anti-PDL1 at a concentration of 50ug/mL, following by goat anti-rabbit Alexa Fluor 647, PDL-1 staining was tested on several lung cancer cell lines: (adenocarcinoma), adenocarcinoma), (squamous) and WBCs asa control. (D) Once validated, patient samples were stained for PD-L1, CK, CD45, DAPI.)
Immunofluorescence (IF)
(Figure 11 Immunofluorescence Validation of PD-L1 intumors in Human Cells (Dhar et al., 2018)(A) Several antibody brands were first tested with RBCs,WBCs, and HeLa cells. (MBS151123) was chosen as it provided the highest staining intensity. (B, C) Using the optimal conditions of anti-PDL1 at a concentration of 50ug/mL, following by goat anti-rabbit Alexa Fluor 647, PDL-1 staining was tested on several lung cancer cell lines: (adenocarcinoma), adenocarcinoma), (squamous) and WBCs asa control. (D) Once validated, patient samples were stained for PD-L1, CK, CD45, DAPI.)
Immunofluoresence
(Figure 12 Immunohistochemistry Validation of PD-L1in Human Tumors (Gadiot et al., 2011)Immunohistochemical analysis of patient tumors labeling PD-L1 with anti-PD-L1 antibodies (MBS151123). Several anti-PD-L1 antibodies were tested for staining, “Only 1 antibody gave no background staining and was competitively blocked by the addition of PD-L1Fc protein (MBS151123)”.)
Immunofluoresence
(Figure 12 Immunohistochemistry Validation of PD-L1in Human Tumors (Gadiot et al., 2011)Immunohistochemical analysis of patient tumors labeling PD-L1 with anti-PD-L1 antibodies (MBS151123). Several anti-PD-L1 antibodies were tested for staining, “Only 1 antibody gave no background staining and was competitively blocked by the addition of PD-L1Fc protein (MBS151123)”.)
NCBI and Uniprot Product Information
Observed: 37kDa
Uniprot Description
CD274: Involved in the costimulatory signal, essential for T- cell proliferation and production of IL10 and IFNG, in an IL2- dependent and a PDCD1-independent manner. Interaction with PDCD1 inhibits T-cell proliferation and cytokine production. Belongs to the immunoglobulin superfamily. BTN/MOG family. 3 isoforms of the human protein are produced by alternative splicing.
Protein type: Membrane protein, integral
Chromosomal Location of Human Ortholog: 9p24
Cellular Component: integral to membrane; plasma membrane; endomembrane system; external side of plasma membrane
Molecular Function: protein binding
Biological Process: regulation of activated T cell proliferation; cell surface receptor linked signal transduction; negative regulation of interleukin-10 production; negative regulation of interferon-gamma production; negative regulation of activated T cell proliferation; T cell costimulation; positive regulation of T cell proliferation; immune response; signal transduction
Research Articles on CD274
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Product Notes
The CD274 cd274 (Catalog #AAA151123) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The PDL-1 Antibody reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's PDL-1 can be used in a range of immunoassay formats including, but not limited to, ELISA (EIA), Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Immunofluorescence (IF), Flow Cytometry (FC). Optimal dilutions for each application to be determined by the researcher. Researchers should empirically determine the suitability of the CD274 cd274 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "PDL-1, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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