Human/Mouse/Rat AIF Antibody

Catalog # Availability Size / Price Qty
AF1457
AF1457-SP
AIF in MCF-7 Human Cell Line.
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Product Details
Citations (6)
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Reviews (1)

Human/Mouse/Rat AIF Antibody Summary

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 either lyophilized or 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
Knockout Validated
AIF is specifically detected in HEK293T human embryonic kidney parental cell line but is not detectable in AIF knockout HEK293T cell line.
 

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

Immunocytochemistry AIF antibody in MCF-7 Human Cell Line by Immunocytochemistry (ICC). View Larger

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.

Western Blot Detection of Human/Mouse/Rat AIF antibody by Western Blot. View Larger

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 antibody in Jurkat Human Cell Line by Immunocytochemistry (ICC). View Larger

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.

Immunocytochemistry AIF antibody in HeLa Human Cell Line by Immunocytochemistry (ICC). View Larger

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.

Simple Western Detection of Human AIF antibody by Simple Western<SUP>TM</SUP>. View Larger

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.

Knockout Validated Western Blot Shows Human AIF Antibody Specificity by Using Knockout Cell Line. View Larger

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

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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

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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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  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, 2017-02-06;292(11):4727-4742.
    Species: Human
    Sample Types: Cell Lysates, Whole Cells
    Applications: ICC, Western Blot
  2. Delivery of a survivin promoter-driven antisense survivin-expressing plasmid DNA as a cancer therapeutic: a proof-of-concept study
    Onco Targets Ther, 2016-05-03;9(0):2601-13.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  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, 2016-04-14;11(4):e0148500.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  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, 2013-04-26;8(4):e60730.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: 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., 2012-05-19;181(1):53-61.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC-Fr
  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., 2011-10-08;179(6):2798-809.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: ICC

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Human/Mouse/Rat AIF Antibody
By Anonymous on 11/21/2022
Application: WB Sample Tested: HepG2 human hepatocellular carcinoma cell line Species: Human

Cell lysate from HepG2 cells, 20 ug loaded. 10% gel, NIR western blot with LICOR secondary antibodies