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Product Info Summary
| SKU: | PB9524 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-BAG2 Antibody Picoband®
SKU/Catalog Number
PB9524
PB0552 is an alternative SKU for this antibody, used in previous lots.
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-BAG2 Antibody Picoband® catalog # PB9524. Tested in Flow Cytometry, IF, ICC, 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-BAG2 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9524)
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 BAG2 recombinant protein (Position: M1-N211). Human BAG2 shares 93.4% amino acid (aa) sequence identity with mouse BAG2.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PB9524 is reactive to BAG2 in Human
Observed Molecular Weight
24 kDa
Calculated molecular weight
23.8 kDa
Background of BAG2
BAG family molecular chaperone regulator 2 is a protein that in humans is encoded by the BAG2 gene. The predicted BAG2 protein contains 211 amino acids. The BAG domains of BAG1, BAG2, and BAG3 interact specifically with the Hsc70 ATPase domain in vitro and in mammalian cells. All 3 proteins bind with high affinity to the ATPase domain of Hsc70 and inhibit its chaperone activity in a Hip-repressible manner. The functional antagonisms displayed between BAG family proteins and Hip suggest that a proper balance of these 2 types of protein is required for achieving optimal cycles of substrate binding and release required for inducting conformational changes in proteins, with Hip promoting peptide substrate binding by Hsc70/Hsp70 and BAG family proteins promoting dissociation.
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
PB9524 is guaranteed for 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, 0.1-0.5μg/ml, Human
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human
Positive Control
WB: human Jurkat whole cell, human Hela whole cell, human HepG2 whole cell, human K562 whole cell
ICC/IF: A431 cell
FCM: THP-1 cell
Validation Images & Assay Conditions
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Flow Cytometry analysis of THP-1 cells using anti-BAG2 antibody (PB9524).
Overlay histogram showing THP-1 cells stained with PB9524 (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-BAG2 Antibody (PB9524, 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 BAG2 using anti-BAG2 antibody (PB9524).
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 Jurkat whole cell lysates,
Lane 2: human Hela whole cell lysates,
Lane 3: human HepG2 whole cell lysates,
Lane 4: human K562 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-BAG2 antigen affinity purified polyclonal antibody (Catalog # PB9524) 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 BAG2 at approximately 24 kDa. The expected band size for BAG2 is at 24 kDa.
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IF analysis of BAG2 using anti-BAG2 antibody (PB9524).
BAG2 was detected in an immunocytochemical section of A431 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-BAG2 Antibody (PB9524) overnight at 4°C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (BA1142) was used as secondary antibody at 1:100 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.
Specific Publications For Anti-BAG2 Antibody Picoband® (PB9524)
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1 Customer Q&As for Anti-BAG2 Antibody Picoband®
Question
We are currently using anti-BAG2 antibody PB9524 for mouse tissue, and we are well pleased with the IHC results. The species of reactivity given in the datasheet says human, mouse, rat. Is it true that the antibody can work on pig tissues as well?
D. Dhar
Verified customer
Asked: 2015-04-17
Answer
The anti-BAG2 antibody (PB9524) has not been validated for cross reactivity specifically with pig 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 pig you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2015-04-17
