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Product Info Summary
| SKU: | PB9915 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | Flow Cytometry, IP, IF, ICC, WB |
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Product info
Product Name
Anti-AFG3L2 Antibody Picoband®
SKU/Catalog Number
PB9915
PB0961 is an alternative SKU for this antibody, used in previous lots.
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-AFG3L2 Antibody Picoband® catalog # PB9915. Tested in Flow Cytometry, IP, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat. 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-AFG3L2 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9915)
Host
Rabbit
Contents
Each vial contains antibody formulated with stabilizing components, 0.9 mg NaCl, 0.2 mg Na2HPO4, and 0.05 mg NaN3.
*This antibody is supplied in a stabilized formulation.
Compatibility with conjugation reactions depends on the chemistry of the conjugation method used.
For conjugation methods that are not compatible with the stabilizing components present in this formulation, a carrier-free antibody format is required.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E. coli-derived human AFG3L2 recombinant protein (Position: R168-D250). Human AFG3L2 shares 100% amino acid (aa) sequence identity with mouse AFG3L2.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PB9915 is reactive to AFG3L2 in Human, Mouse, Rat
Observed Molecular Weight
89 kDa
Calculated molecular weight
88.6 kDa
Background of AFG3L2
AFG3L2 is the catalytic subunit of the m-AAA protease, an ATP-dependent proteolytic complex of the mitochondrial inner membrane that degrades misfolded proteins and regulates ribosome assembly. In humans, it is encoded by the AFG3L2 gene. This gene encodes a protein localized in mitochondria and closely related to paraplegin. The paraplegin gene is responsible for an autosomal recessive form of hereditary spastic paraplegia. And this gene is a candidate gene for other hereditary spastic paraplegias or neurodegenerative disorders as well as spastic ataxia-neuropathy syndrome.
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
PB9915 is guaranteed for Flow Cytometry, IP, 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, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Immunoprecipitation, 0.5-2 μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: human Hela whole cell, human MCF-7 whole cell, human Jurkat whole cell, human MDA-MB-453 whole cell, rat brain tissue, rat liver tissue, mouse brain tissue, mouse liver tissue
ICC/IF: U20S cell
IP: MCF-7 cell
FCM: A431 cell
Validation Images & Assay Conditions
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Western blot analysis of AFG3L2 using anti-AFG3L2 antibody (PB9915).
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 Hela whole cell lysates,
Lane 2: human MCF-7 whole cell lysates,
Lane 3: human Jurkat whole cell lysates,
Lane 4: human MDA-MB-453 whole cell lysates,
Lane 5: rat brain tissue lysates,
Lane 6: rat liver tissue lysates,
Lane 7: mouse brain tissue lysates,
Lane 8: mouse liver tissue 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-AFG3L2 antigen affinity purified polyclonal antibody (Catalog # PB9915) 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 AFG3L2 at approximately 89 kDa. The expected band size for AFG3L2 is at 89 kDa.
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IF analysis of AFG3L2 using anti-AFG3L2 antibody (PB9915).
AFG3L2 was detected in immunocytochemical section of U20S 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 2μg/mL rabbit anti-AFG3L2 Antibody (PB9915) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) 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.
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Immunoprecipitating (IP) AFG3L2 in MCF-7 whole cell lysate.
Western blot analysis of AFG3L2 using anti-AFG3L2 antibody (PB9915);
Lane 1: MCF-7 whole cell lysates (30ug);
Lane 2: Rabbit control IgG instead of anti-AFG3L2 antibody in MCF-7 whole cell lysate;
Lane 3: anti-AFG3L2 antibody (2μg) + MCF-7 whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-AFG3L2 antigen affinity purified polyclonal antibody (PB9915) at a dilution of 0.5 μg/mL and probed with a goat anti-rabbit IgG-HRP secondary antibody (Catalog # BA1054). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1196-200). A specific band was detected for AFG3L2 at approximately 89 kDa. The expected band size for AFG3L2 is at 89 kDa.
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Flow Cytometry analysis of A431 cells using anti-AFG3L2 antibody (PB9915).
Overlay histogram showing A431 cells stained with PB9915 (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-AFG3L2 Antibody (PB9915,1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-AFG3L2 Antibody Picoband® (PB9915)
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4 Customer Q&As for Anti-AFG3L2 Antibody Picoband®
Question
We want to test anti-AFG3L2 antibody PB9915 on rat eye for research purposes, then I may be interested in using anti-AFG3L2 antibody PB9915 for diagnostic purposes as well. Is the antibody suitable for diagnostic purposes?
Verified Customer
Verified customer
Asked: 2020-03-09
Answer
The products we sell, including anti-AFG3L2 antibody PB9915, 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: 2020-03-09
Question
Does PB9915 anti-AFG3L2 antibody work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
C. Li
Verified customer
Asked: 2020-01-03
Answer
It shows on the product datasheet, PB9915 anti-AFG3L2 antibody as been validated on WB. It is best to use PFA for fixation because it has better tissue penetration ability. PFA needs to be prepared fresh before use. Long term stored PFA turns into formalin, as the PFA molecules congregate and become formalin.
Boster Scientific Support
Answered: 2020-01-03
Question
We are currently using anti-AFG3L2 antibody PB9915 for human tissue, and we are happy with the WB results. The species of reactivity given in the datasheet says human, mouse, rat. Is it true that the antibody can work on zebrafish tissues as well?
Verified Customer
Verified customer
Asked: 2019-06-03
Answer
The anti-AFG3L2 antibody (PB9915) has not been validated for cross reactivity specifically with zebrafish tissues, though 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 zebrafish you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2019-06-03
Question
Is a blocking peptide available for product anti-AFG3L2 antibody (PB9915)?
T. Johnson
Verified customer
Asked: 2014-07-10
Answer
We do provide the blocking peptide for product anti-AFG3L2 antibody (PB9915). If you would like to place an order for it please contact support@bosterbio.com and make a special request.
Boster Scientific Support
Answered: 2014-07-10
