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Product Info Summary
| SKU: | A03637-2 |
|---|---|
| Size: | 100 µg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-PAWR Antibody Picoband®
SKU/Catalog Number
A03637-2
Size
100 µg/vial
Form
Lyophilized
Description
Boster Bio Anti-PAWR Antibody Picoband® catalog # A03637-2. Tested in ELISA, IF, ICC, WB, Flow Cytometry 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
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 freezing and thawing.
Cite This Product
Anti-PAWR Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A03637-2)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
IgG
Immunogen
E.coli-derived human PAWR recombinant protein (Position: R160-D255). Human PAWR shares 82.3% and 81.2% amino acid (aa) sequence identity with mouse and rat PAWR, respectively.
Cross-reactivity
No cross reactivity with other proteins.
Reactive Species
A03637-2 is reactive to PAWR in Human
Observed Molecular Weight
41 kDa
Calculated molecular weight
36.6 kDa
Background of PAWR
PRKC apoptosis WT1 regulator protein, or Prostate apoptosis response-4, is a tumor-suppressor protein coded for in the human by the PAWR gene, that induces apoptosis in cancer cells, but not in normal cells. This gene encodes a tumor suppressor protein that selectively induces apoptosis in cancer cells through intracellular and extracellular mechanisms. The intracellular mechanism involves the inhibition of pro-survival pathways and the activation of Fas-mediated apoptosis, while the extracellular mechanism involves the binding of a secreted form of this protein to glucose regulated protein 78 (GRP78) on the cell surface, which leads to activation of the extrinsic apoptotic pathway. This gene is located on the unstable human chromosomal 12q21 region and is often deleted or mutated different tumors. The encoded protein also plays an important role in the progression of age-related diseases.
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
A03637-2 is guaranteed for ELISA, Flow Cytometry, IF, ICC, 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, 0.1-0.25 μg/ml, Human
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry (Fixed), 1-3 μg /1x106 cells, Human
ELISA, 0.1-0.5 μg/ml, -
Positive Control
WB: human SW620 whole cell, human SiHa whole cell, human RT4 whole cell, human Caco-2 whole cell
ICC/IF: HELA cell
FCM: Caco-2 cell
Validation Images & Assay Conditions
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Western blot analysis of PAWR using anti-PAWR antibody (A03637-2).
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 SW620 whole cell lysates,
Lane 2: human SiHa whole cell lysates,
Lane 3: human RT4 whole cell lysates,
Lane 4: human Caco-2 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-PAWR antigen affinity purified polyclonal antibody (Catalog # A03637-2) at 0.25 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 PAWR at approximately 41 kDa. The expected band size for PAWR is at 37 kDa.
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IF analysis of PAWR using anti-PAWR antibody (A03637-2).
PAWR was detected in an immunocytochemical section of HELA cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-PAWR Antibody (A03637-2) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
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Flow Cytometry analysis of Caco-2 cells using anti-PAWR antibody (A03637-2).
Overlay histogram showing Caco-2 cells stained with A03637-2 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PAWR Antibody (A03637-2, 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 (Red line) was also used as a control.
Specific Publications For Anti-PAWR Antibody Picoband® (A03637-2)
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