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Product Info Summary
| SKU: | A03824 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | Flow Cytometry, IP, IHC, WB |
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Product info
Product Name
Anti-AFF4 Antibody Picoband®
SKU/Catalog Number
A03824
Size
100 μg/vial
Form
Lyophilized
Description
AFF4 is commonly described as a scaffold component of the super elongation complex involved in transcriptional elongation control (putative here due to page access failure), with disease-context usage in some leukemia/transcription-dysregulation studies (putative). Assay context: per provided product description, validated for Flow Cytometry, IP, IHC, and Western blot in Human; for IP/IHC workflows, analytical confirmation can be framed within broader assay services method-context. Often interpreted alongside DNA-damage checkpoint signaling such as TP53BP1 (53BP1) (putative transcription–damage response adjacency) and nucleocytoplasmic transport factors such as AGFG1 (putative).
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-AFF4 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A03824)
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
A synthetic peptide corresponding to a sequence at the N-terminus of human AFF4, identical to the related mouse sequence.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A03824 is reactive to AFF4 in Human
Observed Molecular Weight
150 kDa
Calculated molecular weight
127.5 kDa
Background of AFF4
The AFF4 gene encodes a scaffold protein that functions as a core component of the super elongation complex (SEC), which is involved in transcriptional regulation during embryogenesis. The protein encoded by this gene belongs to the AF4 family of transcription factors involved in leukemia. It is a component of the positive transcription elongation factor b (P-TEFb) complex. This gene is mapped to chromosome 5q31.
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
A03824 is guaranteed for Flow Cytometry, IP, IHC, 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
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human, Mouse, Rat
Immunoprecipitation, 0.5-2 μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: human Hela whole cell, human HepG2 whole cell, human 293T whole cell, human U87 whole cell
IHC: human placenta tissue
IP: 293T cell
FCM: SiHa cell
Validation Images & Assay Conditions
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Western blot analysis of AFF4 using anti-AFF4 antibody (A03824).
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 HepG2 whole cell lysates,
Lane 3: human 293T whole cell lysates,
Lane 4: human U87 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-AFF4 antigen affinity purified polyclonal antibody (Catalog # A03824) 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 AFF4 at approximately 150 kDa. The expected band size for AFF4 is at 127 kDa.
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Immunoprecipitating (IP) AFF4 in 293T whole cell lysate.
Western blot analysis of AFF4 using anti-AFF4 antibody (A03824);
Lane 1: 293T whole cell lysates (30ug);
Lane 2: Rabbit control IgG instead of anti-AFF4 antibody in 293T whole cell lysate;
Lane 3: anti-AFF4 antibody (2μg) + 293T whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-AFF4 antigen affinity purified polyclonal antibody (A03824) 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 AFF4 at approximately 150 kDa. The expected band size for AFF4 is at 127 kDa.
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Flow Cytometry analysis of SiHa cells using anti-AFF4 antibody (A03824).
Overlay histogram showing SiHa cells stained with A03824 (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-AFF4 Antibody (A03824,1μg/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.
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IHC analysis of AFF4 using anti-AFF4 antibody (A03824).
AFF4 was detected in a paraffin-embedded section of human placenta tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-AFF4 Antibody (A03824) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Specific Publications For Anti-AFF4 Antibody Picoband® (A03824)
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5 Customer Q&As for Anti-AFF4 Antibody Picoband®
Question
Will anti-AFF4 antibody A03824 work for Flow Cytometry with leukemic t-cell?
Verified Customer
Verified customer
Asked: 2020-02-18
Answer
According to the expression profile of leukemic t-cell, AFF4 is highly expressed in leukemic t-cell. So, it is likely that anti-AFF4 antibody A03824 will work for Flow Cytometry with leukemic t-cell.
Boster Scientific Support
Answered: 2020-02-18
Question
I appreciate helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for leukemic t-cell using anti-AFF4 antibody A03824. Let me know if you need anything else.
Verified Customer
Verified customer
Asked: 2019-10-02
Answer
Thanks for 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: 2019-10-02
Question
Is this A03824 anti-AFF4 antibody reactive to the isotypes of AFF4?
Verified Customer
Verified customer
Asked: 2019-06-19
Answer
The immunogen of A03824 anti-AFF4 antibody is A synthetic peptide corresponding to a sequence at the N-terminus of human AFF4 (6-53aa RNVLRMKERERRNQEIQQGEDAFPPSSPLFAEPYKVTSKEDKLSSRIQ), identical to the related mouse sequence. 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: 2019-06-19
Question
We are currently using anti-AFF4 antibody A03824 for human tissue, and we are satisfied with the WB results. The species of reactivity given in the datasheet says human. Is it likely that the antibody can work on pig tissues as well?
S. Wu
Verified customer
Asked: 2017-08-16
Answer
The anti-AFF4 antibody (A03824) 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: 2017-08-16
Question
Will anti-AFF4 antibody A03824 work on pig IHC-F with brain?
Verified Customer
Verified customer
Asked: 2017-07-20
Answer
Our lab technicians have not tested anti-AFF4 antibody A03824 on pig. You can run a BLAST between pig and the immunogen sequence of anti-AFF4 antibody A03824 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 pig 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 pig brain in IHC-F, you can get your next antibody for free.
Boster Scientific Support
Answered: 2017-07-20
