- PA5-18337 - Invitrogen Antibodies CD274 antibody

Zhou Q, Xiong W, Zhou X, Gao RS, Lin QF, Liu HY, Li JN, Tian XF
Molecular medicine reports 2019 Nov;20(5):4244-4252

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of PD-L1 by a PD-L1 monoclonal antibody (Product # PA5-18337) at a concentration of 0.3 µg/mL. Human Heart (A) lysate + Blocking peptide (B) (35µg protein in RIPA buffer). Detected by chemiluminescence.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot staining of Human Heart lysate using Product # PA5-18337 at a concentration of 0.3 µg/mL, the primary antibody incubation was 1 hour and the detection method was chemiluminescence.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of PDL1 was achieved by transfecting A549 cells with PDL1 specific siRNAs (Silencer® select Product # s26547, s26548). Western blot analysis (Fig. a) was performed using membrane enriched extracts from the PDL1 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with PDL1 Polyclonal Antibody (Product # PA5-18337, 0.1 µg/mL) and Rabbit anti-Goat IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27014, 0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to PDL1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on modified whole cell extracts (1% SDS) (30 µg lysate) of A549 (Lane 1), HeLa (Lane 2), Hep G2 (Lane 3), HCT 116 (Lane 4), A-431 (Lane 5), C2C12 (Lane 6), L6 (Lane 7) and RD (Lane 8). The blot was probed with anti-PD-L1 Polyclonal Antibody (Product # PA5-18337, 1:1000 dilution) and detected by chemiluminescence using Rabbit anti-Goat IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27014, 0.25 µg/mL, 1:4000 dilution). A 50 kDa band corresponding to PD-L1 was detected across the cell lines tested. An additional band of 60kDa was observed across all the cell lines tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of PD-L1 in U2OS cells using a PD-L1 monoclonal antibody (Product # PA5-18337) at 10 µg/mL for1hr. The cells were paraformaldehyde fixed and permeabilized with 0.15% Triton. Primary incubation was followed by Alexa Fluor 488 secondary antibody (2 µg/mL) showing cytoplasmic staining. The nuclear stain is DAPI (blue). Negative control: Unimmunized goat IgG (10 µg/mL)followed by Alexa Fluor 488 secondary antibody (2 µg/mL).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of PD-L1 in A431 cells using a PD-L1 monoclonal antibody (Product # PA5-18337) at 10 µg/mL for1hr. The cells were paraformaldehyde fixed and permeabilized with 0.15% Triton. Primary incubation was followed by Alexa Fluor 488 secondary antibody (2 µg/mL) showing cytoplasmic staining. The nuclear stain is DAPI (blue). Negative control: Unimmunized goat IgG (10 µg/mL)followed by Alexa Fluor 488 secondary antibody (2 µg/mL).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometric analysis of PD-L1 in Jurkat cells using a PD-L1 monoclonal antibody (Product # PA5-18337) at 10 µg/mL for 1hr, depicted by the blue line. The cells were paraformaldehyde fixed and permeabilized with 0.5% Triton. Primary incubation followed by Alexa Fluor 488 secondary antibody (1 µg/mL). IgG control: Unimmunized goat IgG (black line) followed by Alexa Fluor 488 secondary antibody.

PA5-18337