AIF Antibody

Western Blot: AIF Antibody [NBP2-27177] - Western blot shows lysates of HEK293T human embryonic kidney parental cell line and AIF knockout (KO) HEK293T cell line. PVDF membrane was probed with 1:1000 of Rabbit Anti-Human AIF Polyclonal Antibody (Catalog # NBP2-27177) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog #HAF008). Specific band was detected for HDAC1 at approximately 68 kDa (as indicated) in the parental HEK293T cell line, but is not detectable in the knockout HEK293T cell line. This experiment was conducted under reducing conditions.

Western Blot: AIF Antibody [NBP2-27177] - Analysis of AIF in human colon using this antibody. Lane 1, normal colon. Lanes 2-7, colon carcinoma tissue lysatesfrom 6 different patients. mAIF, thought to represent mitochondrial AIF.

Immunohistochemistry-Paraffin: AIF Antibody [NBP2-27177] - Staining of AIF in formalin-fixed, paraffin-embedded normal human colon using this antibody at 1:2000. Hematoxylin-eosin counterstain. In this example, AIF expression is predominant in the upper part of the crypt.

Simple Western: AIF Antibody [NBP2-27177] - Simple Western lane view shows a specific band for AIF in 0.2 mg/ml of Jurkat and MCF-7 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system. *Non-specific interaction with the 230 kDa standard may be seen with this antibody.

• Reactivity: Hu, Mu, Rt, Bv, Ca
• Applications: WB, Simple Western, IHC, IP, KO
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated

AIF Antibody Summary

• Immunogen: (aa 151-170); human
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: AIFM1
• Purity: Unpurified

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry
  • Immunohistochemistry-Frozen 1:1000-1:5000
  • Immunohistochemistry-Paraffin 1:1000-1:5000~
  • Immunoprecipitation 1:50-1:200
  • Knockout Validated
  • Simple Western 1:500
  • Western Blot 1:1000-1:2000~
• Application Notes: In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
  See Simple Western Antibody Database for Simple Western validation: Tested in Jurkat and MCF-7 lysate 0.2 mg/mL, separated by Size, antibody dilution of 1:500

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: 50 ul neat whole antisera
• Preservative: 0.05% Sodium Azide
• Purity: Unpurified

Alternate Names for AIF Antibody

• 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

AIF (apoptosis-inducing factor) was initially discovered as a protein involved in caspase-independent cell death. It is now known that AIF has both vital and lethal functions (reviewed in Modjtahedi et al, 2006). In healthy cells, AIF is a flavoprotein present in the mitochondria where it has vital roles in cellular redox metabolism and mitochondrial bioenergetics. In many models of apoptosis, AIF is released from the mitochondria during mitochondrial outer membrane permeabilization along with other mitochondrial proteins. Upon release, AIF translocates first to the cytosol and then to the nucleus where it induces chromatin condensation and DNA degradation. Although several lines of evidence suggest that AIF is a main mediator of capsase-independent cell death, the mechanisms regulating AIF pro-apoptotic function remain to be fully elucidatied and may depend on the cell type and type of apoptotic stimuli. Human AIF is transcribed from a nuclear gene located on the X chromosome and translated in the cytoplasm to a precursor protein of 613 amino acids (aa) which corresponds to ~67 kDa. The precursor protein is imported into the mitochonria by mitochondrial localization sequences located within the N-terminal prodomain of AIF. Once inside the mitochondria, the prodomain is cleaved giving rise to a mature AIF form of ~57 kDa. AIF isoforms generated from a single AIF gene have been identified, including AIF short isoform 2 (324 aa protein; GenBank no. AAY84739.1) and AIF short isoform 3 (237 aa protein; GenBank noAAY84741.1 (reviewed in Delttre et al, 2006). The generation of multiple isoforms from a common gene is an evolutionary mechanism that increases protein diversiity in eukaryotes. Regulating gene expression through the production of multiple isoforms from a single gene is thought to play a major role in the control of apoptosis and other forms of programmed cell death. It recognizes AIF and AIF isoforms containing the peptide immunogen sequence, RARDPGARVLIVSEDPELP.

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.

Bioinformatics

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