AIF Antibody [Unconjugated]

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.

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.

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 knockoutHEK293T cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

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.

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.

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.

• Reactivity: Hu, Mu, Rt
• Applications: WB, Simple Western, ICC/IF, KO
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: LYOPH

AIF Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human AIF
  Glu121-Asp613
  Accession # O95831
• Specificity: Detects human, mouse, and rat mitochondria-processed AIF.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: AIFM1
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 5-15 ug/mL
  • Knockout Validated
  • Simple Western 1 ug/mL
  • Western Blot 0.1 ug/mL
• Application Notes: In Simple Western only 10-15 uL of the recommended dilution is used per data point.

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for AIF Antibody [Unconjugated]

• 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

Background

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for AIF Antibody (AF1457)(6)

Publications using AF1457

• H Nakajima, M Itakura, T Kubo, A Kaneshige, N Harada, T Izawa, YT Azuma, M Kuwamura, R Yamaji, T Takeuchi Glyceraldehyde-3-phosphate Dehydrogenase (GAPDH) Aggregation Causes Mitochondrial Dysfunction during Oxidative Stress-induced Cell Death J. Biol. Chem, 2017;292(11):4727-4742. 2017 [PMID: 28167533] (Western Blot, Human)
• 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. 2016 [PMID: 27217778] (Western Blot, Human)
• L Admoni-Eli, I Nakdimon, A Shteinfer, T Prezma, T Arif, N Arbel, A Melkov, O Zelichov, I Levi, V Shoshan-Ba Novel Biomarker Proteins in Chronic Lymphocytic Leukemia: Impact on Diagnosis, Prognosis and Treatment PLoS ONE, 2016;11(4):e0148500. 2016 [PMID: 27078856] (Western Blot, Human)
• Hwang J, Hsu C, Yu W, Liu T, Yang W Diet-induced obesity exacerbates auditory degeneration via hypoxia, inflammation, and apoptosis signaling pathways in CD/1 mice. PLoS ONE, 2013;8(4):e60730. 2013 [PMID: 23637762] (IHC, Mouse)
• 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 The regulatory roles of apoptosis-inducing factor in the formation and regression processes of ocular neovascularization. Am. J. Pathol., 2012;181(1):53-61. 2012 [PMID: 22613025] (IHC-Fr, Mouse)
• Notomi S, Hisatomi T, Kanemaru T, Takeda A, Ikeda Y, Enaida H, Kroemer G, Ishibashi T Critical involvement of extracellular ATP acting on P2RX7 purinergic receptors in photoreceptor cell death. Am. J. Pathol., 2011;179(6):2798-809. 2011 [PMID: 21983632] (ICC, Mouse)

Bioinformatics

• Gene Symbol: AIFM1
• Uniprot:
  • O95831(Human)