Product nameHuman CD274 (PD-L1) knockout A549 cell lysate
See all PD-L1 kits
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.
User storage instructions: After reconstitution, store the lysate at -80°C.
Parental Cell LineA549
Mutation descriptionKnockout achieved by using CRISPR/Cas9, 1 bp insertion in exon3.
Knockout validationSanger Sequencing, Western Blot (WB)
Reconstitution notesTo use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.
*Usage of SDS sample buffer is not recommended with these lyophilized lysates.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
Tested applicationsSuitable for: WBmore details
Storage instructionsStore at -80°C. Please refer to protocols.
Components 1 kit Human CD274 knockout A549 cell lysate - IFN gamma treated (48hr, 100 ng/mL) (Lyophilized) 1 x 100µg ab262043 - Human CD274 knockout A549 cell lysate (Lyophilized) 1 x 100µg Human Wild Type A549 cell lysate - IFN gamma treated (48hr, 100 ng/mL) (Lyophilized) 1 x 100µg ab255554 - Human Wild Type A549 cell lysate (Lyophilized) 1 x 100µg
STR AnalysisAmelogenin X,Y D5S818: 11 D13S317: 11 D7S820: 8, 11 D16S539: 11, 12 vWA: 14 TH01: 8,9.3 TPOX: 8,11 CSF1PO: 10, 12
FunctionInvolved 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.
Tissue specificityHighly expressed in the heart, skeletal muscle, placenta and lung. Weakly expressed in the thymus, spleen, kidney and liver. Expressed on activated T- and B-cells, dendritic cells, keratinocytes and monocytes.
Sequence similaritiesBelongs to the immunoglobulin superfamily. BTN/MOG family.
Contains 1 Ig-like C2-type (immunoglobulin-like) domain.
Contains 1 Ig-like V-type (immunoglobulin-like) domain.
Cellular localizationCell membrane and Endomembrane system.
- Information by UniProt
- B7 H
- B7 H1
- B7 homolog 1
KO cell lines
Our Abpromise guarantee covers the use of ab256866 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use at an assay dependent concentration. Predicted molecular weight: 33 kDa.|
Lane 1: Wild-type A549 treated with 100 ng/mL IFN gamma for 48 h cell lysate (20 µg)
Lane 2: CD274 knockout A549 treated with 100 ng/mL IFN gamma for 48 h cell lysate (20 µg)
Lane 3: U-87 MG cell lysate (20 µg)
Lane 4: MCF7 cell lysate (20 µg)
Lane 5: Wild-type A549 untreated cell lysate (20 µg)
Lane 6: CD274 knockout A549 untreated cell lysate (20 µg)
ab213524 Recombinant Anti-PD-L1 antibody [EPR19759] was shown to specifically react with PD-L1 in wild-type A549 treated with 100 ng/mL IFN gamma for 48 h cells in western blot. Loss of signal was observed when both treated and untreated knockout cell line ab267055 (treated and untreated knockout cell lysates ab256866) were used. Wild-type and PD-L1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab213524 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Homozygous: 1 bp insertion in exon3
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab256866 has not yet been referenced specifically in any publications.