Product Info Summary
SKU: | PA1009 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Mouse, Rat |
Host: | Rabbit |
Application: | IF, IHC, IHC-F, ICC, WB |
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Product info
Product Name
Anti-Apoptosis inhibitor 5/API5 Antibody
SKU/Catalog Number
PA1009
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Apoptosis inhibitor 5/API5 Antibody catalog # PA1009. Tested in IF, IHC, IHC-F, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
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-Apoptosis inhibitor 5/API5 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1009)
Host
Rabbit
Contents
Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
A synthetic peptide corresponding to a sequence at the C-terminus of human Apoptosis inhibitor 5, different from the related mouse sequence by one amino acid.
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PA1009 is reactive to API5 in Human, Mouse, Rat
Applications
PA1009 is guaranteed for IF, IHC, IHC-F, ICC, WB Boster Guarantee
Observed Molecular Weight
59 kDa
Calculated molecular weight
59.005kDa
Background of API5
Many growth factors and cytokines act as cellular survival factors by preventing programmed cell death (apoptosis). Apoptosis inhibitor 5 (API5) is an antiapoptotic factor which may have a role in protein assembly. The API5 gene to chromosome 11p12 based on an alignment of the API5 sequence with the genomic sequence It is a critical determinant of E2F1-induced apoptosis, acting downstream of E2F to suppress E2F-dependent apoptosis without generally blocking E2F-dependent transcription.
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.
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Assay dilution & Images
Reconsitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
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, Rat, Mouse
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat, By Heat
Immunohistochemistry (Frozen Section), 0.5-1μg/ml, Rat, Human, Mouse
Immunocytochemistry , 0.5-1μg/ml, Human, Mouse
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of API5 using anti-API5 antibody (PA1009).
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: Rat Cardiac Muscle Tissue Lysate
Lane 2: Rat Brain Tissue Lysate,
Lane 3: Rat Testis Tissue Lysate,
Lane 4: Rat Placenta Tissue Lysate,
Lane 5: MCF-7 Cell Lysate,
Lane 6: HELA Cell Lysate,
Lane 7: CEM Cell Lysate,
Lane 8: SMMC Cell Lysate,
Lane 9: COLO320 Cell Lysate.
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-API5 antigen affinity purified polyclonal antibody (Catalog # PA1009) 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 API5 at approximately 59 kDa. The expected band size for API5 is at 59 kDa.
Click image to see more details
Figure 2. IHC analysis of API5 using anti-API5 antibody (PA1009).
API5 was detected in a paraffin-embedded section of Rat Brain 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 1 μg/ml rabbit anti-API5 Antibody (PA1009) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 3. ICC analysis of API5 using anti-API5 antibody (PA1009).
API5 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 1 μg/ml rabbit anti-API5 Antibody (PA1009) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 4. IHC analysis of API5 using anti-API5 antibody (PA1009).
API5 was detected in a frozen section of Rat Cardiac Muscle tissue. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-API5 Antibody (PA1009) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 5. IHC analysis of API5 using anti-API5 antibody (PA1009).
API5 was detected in paraffin-embedded section of human placenta tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-API5 Antibody (PA1009) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 6. IHC analysis of API5 using anti-API5 antibody (PA1009).
API5 was detected in paraffin-embedded section of rat intestines tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-API5 Antibody (PA1009) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 7. IHC analysis of API5 using anti-API5 antibody (PA1009).
API5 was detected in paraffin-embedded section of mouse intestines tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-API5 Antibody (PA1009) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 8. IF analysis of API5 using anti-API5 antibody (PA1009).
API5 was detected in an immunocytochemical section of SK-OV-3 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-API5 Antibody (PA1009) 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.
Protein Target Info & Infographic
Gene/Protein Information For API5 (Source: Uniprot.org, NCBI)
Gene Name
API5
Full Name
Apoptosis inhibitor 5
Weight
59.005kDa
Superfamily
API5 family
Alternative Names
AAC-11API5L1; AAC11fibroblast growth factor 2-interacting factor 2; Antiapoptosis clone 11 protein; API-5; apoptosis inhibitor 5; Cell migration-inducing gene 8 protein; Fibroblast growth factor 2-interacting factor; FIF; migration-inducing protein MIG8; Protein XAGL API5 AAC-11, AAC11 apoptosis inhibitor 5 apoptosis inhibitor 5|FIF|antiapoptosis clone 11 protein|cell migration-inducing gene 8 protein|fibroblast growth factor 2-interacting factor 2|migration-inducing protein MIG8
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on API5, check out the API5 Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for API5: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-Apoptosis inhibitor 5/API5 Antibody (PA1009)
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Customer Q&As
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6 Customer Q&As for Anti-Apoptosis inhibitor 5/API5 Antibody
Question
Would PA1009 anti-Apoptosis inhibitor 5/API5 antibody work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
Verified Customer
Verified customer
Asked: 2020-01-28
Answer
You can see on the product datasheet, PA1009 anti-Apoptosis inhibitor 5/API5 antibody as been validated on IHC-F. 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: 2020-01-28
Question
We are currently using anti-Apoptosis inhibitor 5/API5 antibody PA1009 for mouse tissue, and we are well pleased with the IHC-F results. The species of reactivity given in the datasheet says human, mouse, rat. Is it likely that the antibody can work on pig tissues as well?
Verified Customer
Verified customer
Asked: 2019-08-02
Answer
The anti-Apoptosis inhibitor 5/API5 antibody (PA1009) 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: 2019-08-02
Question
Is this PA1009 anti-Apoptosis inhibitor 5/API5 antibody reactive to the isotypes of API5?
B. Parker
Verified customer
Asked: 2018-11-15
Answer
The immunogen of PA1009 anti-Apoptosis inhibitor 5/API5 antibody is A synthetic peptide corresponding to a sequence at the C-terminus of human Apoptosis inhibitor 5(487-504aa KYSSNLGNFNYERSLQGK)(b), different from the related mouse sequence by one amino acid. 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: 2018-11-15
Question
Does anti-Apoptosis inhibitor 5/API5 antibody PA1009 work on primate IHC-F with ovarian carcinoma?
Verified Customer
Verified customer
Asked: 2018-11-14
Answer
Our lab technicians have not validated anti-Apoptosis inhibitor 5/API5 antibody PA1009 on primate. You can run a BLAST between primate and the immunogen sequence of anti-Apoptosis inhibitor 5/API5 antibody PA1009 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 primate 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 primate ovarian carcinoma in IHC-F, you can get your next antibody for free.
Boster Scientific Support
Answered: 2018-11-14
Question
I have attached the WB image, lot number and protocol we used for colon carcinoma using anti-Apoptosis inhibitor 5/API5 antibody PA1009. Please let me know if you require anything else.
Verified Customer
Verified customer
Asked: 2018-04-16
Answer
Thank you very much for the data. Our lab team are working to resolve this as quickly as possible, and we appreciate your patience and understanding! You have provided everything we needed. Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2018-04-16
Question
Do you have a BSA free version of anti-Apoptosis inhibitor 5/API5 antibody PA1009 available?
Verified Customer
Verified customer
Asked: 2017-10-24
Answer
I appreciate your recent telephone inquiry. I can confirm that some lots of this anti-Apoptosis inhibitor 5/API5 antibody PA1009 are BSA free. For now, these lots are available and we can make a BSA free formula for you free of charge. It will take 3 extra days to prepare. If you require this antibody BSA free again in future, please do not hesitate to contact me and I will be pleased to check which lots we have in stock that are BSA free.
Boster Scientific Support
Answered: 2017-10-24