|Product Name||Anti-PD1/PDCD1 Antibody|
|Storage & Handling||At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.|
|Description||Rabbit IgG polyclonal antibody for Programmed cell death protein 1(PDCD1) detection. Tested with WB in Human;Mouse.|
|Cite This Product||Anti-PD1/PDCD1 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # RP1039)|
|Immunogen||E.coli-derived human PD1 recombinant protein (Position: P101-L288). Human PD1 shares 59% amino acid (aa) sequence identity with mouse PD1.|
Assay Dilutions Overview
Western blot, 0.1-0.5μg/ml, Human, Mouse
Boster's Secondary Antibodies And IHC, WB Kits
The following reagents are used to generate the images below.Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot.
Images And Assay Conditions
Figure 1. Western blot analysis of PD1/PDCD1 using anti- PD1/PDCD1 antibody (RP1039).
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 50ug of sample under reducing conditions.
Lane 1: human Caco-2 whole cell lysates,
Lane 2: human SW620 whole cell lysates,
Lane 3: human COLO-320 whole cell lysates,
Lane 4: human K562 whole cell lysates,
Lane 5: mouse thymus tissue lysates,
Lane 6: mouse RAW246.7 whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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- PD1/PDCD1 antigen affinity purified polyclonal antibody (Catalog # RP1039) 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 1:10000 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 PD1/PDCD1 at approximately 65KD. The expected band size for PD1/PDCD1 is at 32KD.
Protein Target Info (Source: Uniprot.org)
|Protein Name||Programmed cell death protein 1|
|Alternative Names||Programmed cell death protein 1;Protein PD-1;hPD-1;CD279;PDCD1;PD1;|
|Subcellular Localization||Membrane; Single-pass type I membrane protein.|
|Molecular Weight||31647 MW|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|Protein Function||Inhibitory cell surface receptor involved in the regulation of T-cell function during immunity and tolerance. Upon ligand binding, inhibits T-cell effector functions in an antigen- specific manner. Possible cell death inducer, in association with other factors. .|
|Research Areas||Apoptosis, Associated Proteins, Cell Biology, Developmental Biology, Intracellular, Neurogenesis, Neurology Process, Neuroscience, Organogenesis, Protein Phosphorylation, Ser / Thr Kinases, Signal Transduction
*You can search these to find other products in these research areas.
|Background||PDCD1(Programmed cell death 1), also called PD1, encodes a cell surface receptor that is a member of the B7 superfamily involved in immunomodulation. This gene is mapped to 2q37.3. PDCD1 acts as an inhibitory molecule on T cells after interacting with its ligands PDL1 and PDL2. The PDCD1 gene contains 5 exons. This protein is expressed in pro-B-cells and is thought to play a role in their differentiation. Using flow cytometric analysis, It has been found that expression of PDCD1 was upregulated on CD16-positive and CD16-negative monocytes, but not on dendritic cells, in viremic HIV-positive patients, but not in highly active antiretroviral therapy (HAART)-treated HIV-positive patients. PDCD1 upregulation in monocytes was induced by microbial Toll-like receptor ligands and inflammatory cytokines.|
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1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected
2. Post-translational cleavage: this can cause smaller bands and or multiple bands
3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody.
4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.
5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher.,