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Product Info Summary
| SKU: | PB9154 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | Flow Cytometry, WB |
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Product info
Product Name
Anti-PD-L1/CD274 Antibody Picoband®
SKU/Catalog Number
PB9154
PB0166 is an alternative SKU for this antibody, used in previous lots.
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-PD-L1/CD274 Antibody Picoband® catalog # PB9154. Tested in Flow Cytometry, WB applications. This antibody reacts with Human. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-PD-L1/CD274 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9154)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human PD-L1 recombinant protein (Position: E45-T290). Human PD-L1 shares 69% amino acid (aa) sequence identity with mouse PD-L1.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PB9154 is reactive to CD274 in Human
Observed Molecular Weight
40-50 kDa
Calculated molecular weight
33.3 kDa
Background of CD274
Programmed death-ligand 1 (PD-L1) also known as CD274 or B7-H1 is a protein that in humans is encoded by the CD274 gene. It is mapped to 9p24.1. PD-L1 is a 40kDa type 1 transmembrane protein that has been speculated to play a major role in suppressing the immune system during particular events such as pregnancy, tissue allografts, autoimmune disease and other disease states such as hepatitis. It has been concluded that upregulation of PD-L1 on tumor MDCs downregulates T-cell immunity and that PD-L1 blockade may represent an approach for cancer immunotherapy. PD-L1 can provide positive costimulatory signals for innate and adaptive immunity and for protection against intracellular bacterial infection. What’s more, It has been found that PD1/PDL1 pathway may be a good target for restoring antitumor immunity in ovarian cancer.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
PB9154 is guaranteed for Flow Cytometry, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.25-0.5μg/ml, Human
Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human
Positive Control
WB: human PC-3 whole cell, human placenta tissue, human HepG2 whole cell, human THP-1 whole cell, human K562 whole cell, human A549 whole cell, human Hela whole cell
FCM: JK cell
Validation Images & Assay Conditions
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Flow Cytometry analysis of JK cells using anti-PD-L1 antibody (PB9154).
Overlay histogram showing JK cells stained with PB9154 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-PD-L1 Antibody (PB9154, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
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Western blot analysis of PD-L1 using anti-PD-L1 antibody (PB9154).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human PC-3 whole cell lysates,
Lane 2: human placenta tissue lysates,
Lane 3: human HepG2 whole cell lysates,
Lane 4: human THP-1 whole cell lysates,
Lane 5: human K562 whole cell lysates,
Lane 6: human A549 whole cell lysates,
Lane 7: human Hela whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-PD-L1 antigen affinity purified polyclonal antibody (Catalog # PB9154) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for PD-L1 at approximately 40-50 kDa. The expected band size for PD-L1 is at 33 kDa.
Specific Publications For Anti-PD-L1/CD274 Antibody Picoband® (PB9154)
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Customer Q&As
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12 Customer Q&As for Anti-PD-L1/CD274 Antibody Picoband®
Question
My colleagues were content with the WB result of your anti-PD-L1/CD274 antibody. However we have observed positive staining in gastric carcinoma cell membrane using this antibody. Is that expected? Could you tell me where is CD274 supposed to be expressed?
Verified Customer
Verified customer
Asked: 2020-04-28
Answer
According to literature, gastric carcinoma does express CD274. Generally CD274 expresses in cell membrane. Regarding which tissues have CD274 expression, here are a few articles citing expression in various tissues:
Liver, Pubmed ID: 19159218
Lung, Pubmed ID: 15489334
Placenta, Pubmed ID: 11015443, 14702039
Boster Scientific Support
Answered: 2020-04-28
Question
We are currently using anti-PD-L1/CD274 antibody PB9154 for human tissue, and we are happy with the WB results. The species of reactivity given in the datasheet says human. Is it true that the antibody can work on horse tissues as well?
Verified Customer
Verified customer
Asked: 2020-01-08
Answer
The anti-PD-L1/CD274 antibody (PB9154) has not been validated for cross reactivity specifically with horse tissues, but there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in horse you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2020-01-08
Question
Here is the WB image, lot number and protocol we used for liver using anti-PD-L1/CD274 antibody PB9154. Please let me know if you require anything else.
Verified Customer
Verified customer
Asked: 2019-09-03
Answer
Thank you very much for the data. Our lab team are working to resolve this as quickly as possible, and we appreciate your patience and understanding! You have provided everything we needed. Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2019-09-03
Question
Is this PB9154 anti-PD-L1/CD274 antibody reactive to the isotypes of CD274?
Verified Customer
Verified customer
Asked: 2019-06-18
Answer
The immunogen of PB9154 anti-PD-L1/CD274 antibody is E.coli-derived human PD-L1 recombinant protein (Position: E45-T290). Human PD-L1 shares 69% amino acid (aa) sequence identity with mouse PD-L1. Could you tell me which isotype you are interested in so I can help see if the immunogen is part of this isotype?
Boster Scientific Support
Answered: 2019-06-18
Question
Please suggest the suitable negative control tissue for your product Anti-PD-L1/CD274 Antibody Picoband for IHC applications.
Verified Customer
Verified customer
Asked: 2018-07-26
Answer
PD-L1 expressions varies by tumor type so screening is recommended to find positive and negative tumor controls. You can use endothelial cells as negative internal controls within a positive (tumor-) tissue but remember that many tumor specimens have some inflammatory macrophages and tumor associated immune cells which are expected to be positive. That
Boster Scientific Support
Answered: 2018-07-26
Question
Does anti-PD-L1/CD274 antibody PB9154 work for WB with liver?
Verified Customer
Verified customer
Asked: 2018-05-21
Answer
According to the expression profile of liver, CD274 is highly expressed in liver. So, it is likely that anti-PD-L1/CD274 antibody PB9154 will work for WB with liver.
Boster Scientific Support
Answered: 2018-05-21
Question
We are interested in to test anti-PD-L1/CD274 antibody PB9154 on human liver for research purposes, then I may be interested in using anti-PD-L1/CD274 antibody PB9154 for diagnostic purposes as well. Is the antibody suitable for diagnostic purposes?
J. Williams
Verified customer
Asked: 2018-02-26
Answer
The products we sell, including anti-PD-L1/CD274 antibody PB9154, are only intended for research use. They would not be suitable for use in diagnostic work. If you have the means to develop a product into diagnostic use, and are interested in collaborating with us and develop our product into an IVD product, please contact us for more discussions.
Boster Scientific Support
Answered: 2018-02-26
Question
I appreciate helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for liver using anti-PD-L1/CD274 antibody PB9154. Let me know if you need anything else.
Verified Customer
Verified customer
Asked: 2018-01-24
Answer
We appreciate the data. You have provided everything we needed. Our lab team are working to resolve your inquiry as quickly as possible, and we appreciate your patience and understanding! Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2018-01-24
Question
Does anti-PD-L1/CD274 antibody PB9154 work on zebrafish WB with liver?
Verified Customer
Verified customer
Asked: 2017-11-21
Answer
Our lab technicians have not validated anti-PD-L1/CD274 antibody PB9154 on zebrafish. You can run a BLAST between zebrafish and the immunogen sequence of anti-PD-L1/CD274 antibody PB9154 to see if they may cross-react. If the sequence homology is close, then you can perform a pilot test. Keep in mind that since we have not validated zebrafish samples, this use of the antibody is not covered by our guarantee. However we have an innovator award program that if you test this antibody and show it works in zebrafish liver in WB, you can get your next antibody for free.
Boster Scientific Support
Answered: 2017-11-21
Question
I was wanting to use your anti-PD-L1/CD274 antibody for WB for human liver on frozen tissues, but I want to know if it has been tested for this particular application. Has this antibody been tested and is this antibody a good choice for human liver identification?
Verified Customer
Verified customer
Asked: 2017-05-23
Answer
You can see on the product datasheet, PB9154 anti-PD-L1/CD274 antibody has been validated for WB on human tissues. We have an innovator award program that if you test this antibody and show it works in human liver in IHC-frozen, you can get your next antibody for free.
Boster Scientific Support
Answered: 2017-05-23
Question
We have seen staining in human liver. Do you have any suggestions? Is anti-PD-L1/CD274 antibody supposed to stain liver positively?
M. Miller
Verified customer
Asked: 2015-05-22
Answer
Based on literature liver does express CD274. Based on Uniprot.org, CD274 is expressed in chorionic villus, placenta, gastric carcinoma, lung, liver, among other tissues. Regarding which tissues have CD274 expression, here are a few articles citing expression in various tissues:
Liver, Pubmed ID: 19159218
Lung, Pubmed ID: 15489334
Placenta, Pubmed ID: 11015443, 14702039
Boster Scientific Support
Answered: 2015-05-22
Question
My question regards using your anti-PD-L1/CD274 antibody for positive regulation of t cell proliferation studies. Has this antibody been tested with western blotting on hepg2 whole cell lysate? We would like to see some validation images before ordering.
C. Banerjee
Verified customer
Asked: 2015-01-22
Answer
We appreciate your inquiry. This PB9154 anti-PD-L1/CD274 antibody is tested on hepg2 whole cell lysate. It is guaranteed to work for WB in human. Our Boster guarantee will cover your intended experiment even if the sample type has not been be directly tested.
Boster Scientific Support
Answered: 2015-01-22
