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Product Info Summary
| SKU: | PA2232 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | IHC, ICC, WB |
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Product info
Product Name
Anti-AIF/AIFM1 Antibody Picoband®
SKU/Catalog Number
PA2232
BA3715 is an alternative SKU for this antibody, used in previous lots.
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-AIF/AIFM1 Antibody catalog # PA2232. Tested in IHC, 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-AIF/AIFM1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA2232)
Host
Rabbit
Contents
Each vial contains antibody formulated with stabilizing components, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg 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 C-terminus of human AIF, identical to the related rat and mouse sequences.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PA2232 is reactive to AIFM1 in Human, Mouse, Rat
Observed Molecular Weight
67 kDa
Calculated molecular weight
66.9 kDa
Background of AIFM1
Apoptosis-inducing factor 1, mitochondrial, also known as AIF or PDCD8 is a protein that in humans is encoded by the AIFM1 gene. AIFM1 gene is mapped to Xq26.1 based on an alignment of the AIFM1 sequence with the genomic sequence. This gene encodes a flavoprotein essential for nuclear disassembly in apoptotic cells, and it is found in the mitochondrial intermembrane space in healthy cells. Induction of apoptosis results in the translocation of this protein to the nucleus where it affects chromosome condensation and fragmentation. In addition, this gene product induces mitochondria to release the apoptogenic proteins cytochrome c and caspase-9. Mutations in this gene cause combined oxidative phosphorylation deficiency 6, which results in a severe mitochondrial encephalomyopathy. A related pseudogene has been identified on chromosome 10.
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
PA2232 is guaranteed for IHC, 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.
Immunocytochemistry , 0.5-1μg/ml, Human, Mouse, Rat
Immunohistochemistry (Frozen Section), 0.5-1μg/ml, Rat, Human, Mouse
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Rat, Mouse
Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat
Positive Control
WB: human MCF-7 whole cell, human K562 whole cell, human A549 whole cell, human HepG2 whole cell, rat kidney tissue, rat spleen tissue, mouse kidney tissue, mouse spleen tissue
IHC: Human Lung Cancer Tissue, Rat Cardiac Muscle Tissue
ICC/IF: MCF-7 cells
Validation Images & Assay Conditions
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Western blot analysis of AIFM1 using anti-AIFM1 antibody (PA2232).
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 MCF-7 whole cell lysates,
Lane 2: human K562 whole cell lysates,
Lane 3: human A549 whole cell lysates,
Lane 4: human HepG2 whole cell lysates,
Lane 5: rat kidney tissue lysates,
Lane 6: rat spleen tissue lysates,
Lane 7: mouse kidney tissue lysates,
Lane 8: mouse spleen 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-AIFM1 antigen affinity purified polyclonal antibody (Catalog # PA2232) 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 AIFM1 at approximately 67 kDa. The expected band size for AIFM1 is at 67 kDa.
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Anti-AIF antibody, PA2232, IHC(P)
IHC(P): Human Lung Cancer Tissue
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Anti-AIF antibody, PA2232, IHC(P)
IHC(P): Rat Cardiac Muscle Tissue
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IHC analysis of AIF/AIFM1 using anti-AIF/AIFM1 antibody (PA2232).
AIF/AIFM1 was detected in a paraffin-embedded section of human kidney 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-AIF/AIFM1 Antibody (PA2232) 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.
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IF analysis of AIFM1 using anti-AIFM1 antibody (PA2232).
AIFM1 was detected in an immunocytochemical section of MCF-7 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-AIFM1 Antibody (PA2232) 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.
Specific Publications For Anti-AIF/AIFM1 Antibody Picoband® (PA2232)
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5 Customer Q&As for Anti-AIF/AIFM1 Antibody Picoband®
Question
We have seen staining in mouse cervix carcinoma. Are there any suggestions? Is anti-AIF/AIFM1 antibody supposed to stain cervix carcinoma positively?
Verified Customer
Verified customer
Asked: 2019-12-27
Answer
From what I have seen in literature cervix carcinoma does express AIFM1. From what I have seen in Uniprot.org, AIFM1 is expressed in apex of heart, kidney, brain, cervix carcinoma, cervix carcinoma erythroleukemia, liver, among other tissues. Regarding which tissues have AIFM1 expression, here are a few articles citing expression in various tissues:
Brain, Pubmed ID: 10913597, 15489334
Cervix carcinoma, Pubmed ID: 18669648, 18691976
Cervix carcinoma, and Erythroleukemia, Pubmed ID: 23186163
Kidney, Pubmed ID: 14702039
Liver, Pubmed ID: 24275569
Boster Scientific Support
Answered: 2019-12-27
Question
We purchased anti-AIF/AIFM1 antibody for IHC on kidney a few years ago. I am using mouse, and We are going to use the antibody for WB next. I am looking for examining kidney as well as cervix carcinoma in our next experiment. Do you have any suggestion on which antibody would work the best for WB?
Verified Customer
Verified customer
Asked: 2019-12-11
Answer
I looked at the website and datasheets of our anti-AIF/AIFM1 antibody and I see that PA2232 has been validated on mouse in both IHC and WB. Thus PA2232 should work for your application. Our Boster satisfaction guarantee will cover this product for WB in mouse even if the specific tissue type has not been validated. We do have a comprehensive range of products for WB detection and you can check out our website bosterbio.com to find out more information about them.
Boster Scientific Support
Answered: 2019-12-11
Question
My boss were happy with the WB result of your anti-AIF/AIFM1 antibody. However we have seen positive staining in cervix carcinoma erythroleukemia isoform 3: mitochondrion intermembrane space using this antibody. Is that expected? Could you tell me where is AIFM1 supposed to be expressed?
M. Huang
Verified customer
Asked: 2019-09-17
Answer
From literature, cervix carcinoma erythroleukemia does express AIFM1. Generally AIFM1 expresses in mitochondrion intermembrane space., isoform 3: mitochondrion intermembrane space, isoform 5: cytoplasm. Regarding which tissues have AIFM1 expression, here are a few articles citing expression in various tissues:
Brain, Pubmed ID: 10913597, 15489334
Cervix carcinoma, Pubmed ID: 18669648, 18691976
Cervix carcinoma, and Erythroleukemia, Pubmed ID: 23186163
Kidney, Pubmed ID: 14702039
Liver, Pubmed ID: 24275569
Boster Scientific Support
Answered: 2019-09-17
Question
Our lab want to know about using your anti-AIF/AIFM1 antibody for cellular response to oxygen-glucose deprivation studies. Has this antibody been tested with western blotting on tissue lysate? We would like to see some validation images before ordering.
W. Rodriguez
Verified customer
Asked: 2019-04-29
Answer
We appreciate your inquiry. This PA2232 anti-AIF/AIFM1 antibody is validated on lung cancer tissue, tissue lysate, rat heart tissue, cardiac muscle tissue, brain tissue, k562 cell lysate, hepg2 cell lysate, a431 cell lysate, nih3t3 cell lysate. It is guaranteed to work for IHC, ICC, WB in human, mouse, rat. Our Boster guarantee will cover your intended experiment even if the sample type has not been be directly tested.
Boster Scientific Support
Answered: 2019-04-29
Question
We are currently using anti-AIF/AIFM1 antibody PA2232 for mouse tissue, and we are happy 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 feline tissues as well?
C. Miller
Verified customer
Asked: 2019-04-09
Answer
The anti-AIF/AIFM1 antibody (PA2232) has not been validated for cross reactivity specifically with feline 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 feline you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2019-04-09
