Human/Mouse/Rat AIF Antibody

(6 citations)   
  • Species Reactivity
    Human, Mouse, Rat
  • Specificity
    Detects human, mouse, and rat mitochondria-processed AIF.
  • Source
    Polyclonal Rabbit IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human AIF
    Glu121-Asp613
    Accession # O95831
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    See below
  • Simple Western
    1 µg/mL
    See below
  • Immunocytochemistry
    5-15 µg/mL
    See below
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.
Data Examples
Immunocytochemistry
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.
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.
Immunocytochemistry
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 Northern­Lights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (yellow; Catalog # NL004) and counter­stained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Immunocytochemistry
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.
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.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 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.
  • Long Name:
    Apoptosis Inducing Factor
  • Entrez Gene IDs:
    9131 (Human); 26926 (Mouse); 83833 (Rat)
  • Alternate Names:
    AIF; AIFapoptosis-inducing factor 1, mitochondrial; AIFM1; apoptosis-inducing factor, mitochondrion-associated, 1; COXPD6; MGC111425; PDCD8; PDCD8striatal apoptosis-inducing factor; programmed cell death 8 (apoptosis-inducing factor); Programmed cell death protein 8
Related Research Areas
Citations:

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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Species
Applications
Sample Type
  1. 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;292(11):4727-4742.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  2. Delivery of a survivin promoter-driven antisense survivin-expressing plasmid DNA as a cancer therapeutic: a proof-of-concept study
    Onco Targets Ther, 2016;9(0):2601-13.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  3. 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;11(4):e0148500.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  4. 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;8(4):e60730.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC
  5. 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;181(1):53-61.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC Frozen
  6. 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;179(6):2798-809.
    Species: Mouse
    Sample Type: Whole Cells
    Application: ICC
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