Human/Mouse/Rat AIF Antibody Summary
Glu121-Asp613
Accession # O95831
Applications
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Scientific Data
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AIF in MCF-7 Human Cell Line. Apoptosis Inducing Factor (AIF) was detected in immersion fixed MCF-7 human breast cancer cell line using Rabbit Anti-Human/Mouse/Rat AIF Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1457) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 493-conjugated Anti-Rabbit IgG Secondary Antibody (green; Catalog # NL006) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
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Detection of Human/Mouse/Rat AIF by Western Blot. Western blot shows lysates of MCF-7 human breast cancer cell line, Jurkat human acute T cell leukemia cell line, CTLL-2 mouse cytotoxic T cell line, DA3 mouse myeloma cell line, and PC-12 rat adrenal pheochromocytoma cell line. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-Human/Mouse/Rat AIF Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1457) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for AIF at approximately 67 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
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AIF in Jurkat Human Cell Line. AIF was detected in immersion fixed staurosporine-stimulated Jurkat human acute T cell leukemia cell line using Rabbit Anti-Human/Mouse/Rat AIF Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1457) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the Northern-Lights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (yellow; Catalog # NL004) and counter-stained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
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AIF in HeLa Human Cell Line. AIF was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Rabbit Anti-Human/Mouse/Rat AIF Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1457) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to mitochondria. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
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Detection of Human AIF by Simple WesternTM. Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line and MCF‑7 human breast cancer cell line, loaded at 0.2 mg/mL. A specific band was detected for AIF at approximately 69 kDa (as indicated) using 1 µg/mL of Rabbit Anti-Human/Mouse/Rat AIF Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1457). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
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Western Blot Shows Human AIF Specificity by Using Knockout Cell Line. Western blot shows lysates of HEK293T human embryonic kidney parental cell line and AIF knockout HEK293T cell line (KO). PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-Human/Mouse/Rat AIF Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1457) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for AIF at approximately 65 kDa (as indicated) in the parental HEK293T cell line, but is not detectable in knockout HEK293T cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Reconstitution Calculator
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: AIF
Apoptosis-inducing factor (AIF, also known as programmed cell death protein 8) is a 58 kDa member of the FAD-dependent oxidoreductase family of molecules. It is ubiquitously expressed and found in the mitochondrial intermembrane space. AIF likely acts as a mitochondrial antioxidant providing protection via NADH oxidase activity. Upon release from the mitochondria, AIF passes into the nucleus where it initiates apoptosis. Human AIF precursor is 67 kDa in size and 613 amino acids (aa) in length and contains a cleavable N-terminal 102 aa mitochondrial localization sequence, followed by a spacer region (aa 103‑129) and an oxidoreductase domain (aa 130‑613) that possesses an NLS (aa 446‑451). Over aa 121‑613, human AIF shares 95% aa identity with mouse AIF.
Product Datasheets
Citations for Human/Mouse/Rat AIF Antibody
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
6
Citations: Showing 1 - 6
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Glyceraldehyde-3-phosphate Dehydrogenase (GAPDH) Aggregation Causes Mitochondrial Dysfunction during Oxidative Stress-induced Cell Death
Authors: H Nakajima, M Itakura, T Kubo, A Kaneshige, N Harada, T Izawa, YT Azuma, M Kuwamura, R Yamaji, T Takeuchi
J. Biol. Chem, 2017-02-06;292(11):4727-4742.
Species: Human
Sample Types: Cell Lysates, Whole Cells
Applications: ICC, Western Blot -
Delivery of a survivin promoter-driven antisense survivin-expressing plasmid DNA as a cancer therapeutic: a proof-of-concept study
Onco Targets Ther, 2016-05-03;9(0):2601-13.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot -
Novel Biomarker Proteins in Chronic Lymphocytic Leukemia: Impact on Diagnosis, Prognosis and Treatment
Authors: L Admoni-Eli, I Nakdimon, A Shteinfer, T Prezma, T Arif, N Arbel, A Melkov, O Zelichov, I Levi, V Shoshan-Ba
PLoS ONE, 2016-04-14;11(4):e0148500.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot -
Diet-induced obesity exacerbates auditory degeneration via hypoxia, inflammation, and apoptosis signaling pathways in CD/1 mice.
Authors: Hwang J, Hsu C, Yu W, Liu T, Yang W
PLoS ONE, 2013-04-26;8(4):e60730.
Species: Mouse
Sample Types: Whole Tissue
Applications: IHC -
The regulatory roles of apoptosis-inducing factor in the formation and regression processes of ocular neovascularization.
Authors: Hisatomi T, Nakao S, Murakami Y, Noda K, Nakazawa T, Notomi S, Connolly E, She H, Almulki L, Ito Y, Vavvas DG, Ishibashi T, Miller JW
Am. J. Pathol., 2012-05-19;181(1):53-61.
Species: Mouse
Sample Types: Whole Tissue
Applications: IHC-Fr -
Critical involvement of extracellular ATP acting on P2RX7 purinergic receptors in photoreceptor cell death.
Authors: Notomi S, Hisatomi T, Kanemaru T, Takeda A, Ikeda Y, Enaida H, Kroemer G, Ishibashi T
Am. J. Pathol., 2011-10-08;179(6):2798-809.
Species: Mouse
Sample Types: Whole Cells
Applications: ICC
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Cell lysate from HepG2 cells, 20 ug loaded. 10% gel, NIR western blot with LICOR secondary antibodies
