PRODUCT CODE: ET1603-4

AIF Recombinant Rabbit Monoclonal Antibody [SZ05-01] (ET1603-4)

  • Recombinant

Applications

  • WB

  • ICC

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

Western blot analysis of AIF on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1603-4, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: SKOV-3 cell lysate<br />
Lane 2: Hela cell lysate
  • Western blot analysis of AIF on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1603-4, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: SKOV-3 cell lysate<br />
Lane 2: Hela cell lysate
  • ICC staining of AIF in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1603-4, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of AIF in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1603-4, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-AIF antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1603-4, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-AIF antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1603-4, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded mouse liver tissue using anti-AIF antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1603-4, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded mouse heart tissue using anti-AIF antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1603-4, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Flow cytometric analysis of AIF was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1603-4, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of AIF on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1603-4, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: SKOV-3 cell lysate
Lane 2: Hela cell lysate

Applications

  • WB

  • ICC

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

AIF Recombinant Rabbit Monoclonal Antibody [SZ05-01] (ET1603-4)

Immunogen

Synthetic peptide within human aif aa 500-550.

Host

Rabbit

Positive Control

SKOV-3 cell lysate, Hela cell lysate, Hela, HepG2, human liver tissue, human kidney tissue, mouse liver tissue, mouse heart tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SZ05-01

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

67 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:500-1:5,000

  • ICC

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

  • IP

  • assay-dependent

TARGET

UNIPROT #

PROTEIN NAME

AIF

SYNONYMS

AIFM1 antibody; AIFM1_HUMAN antibody; Apoptosis inducing factor 1, mitochondrial antibody; Apoptosis inducing factor antibody; Apoptosis inducing factor, mitochondrion associated, 1 antibody; Apoptosis-inducing factor 1 antibody; CMTX4 antibody; COWCK antibody; COXPD6 antibody; Harlequin antibody; Hq antibody; mAIF antibody; MGC111425 antibody; MGC5706 antibody; mitochondrial antibody; Neuropathy, axonal motor-sensory, with deafness and mental retardation antibody; neuropathy, axonal, motor-sensory with deafness and mental retardation (Cowchock syndrome) antibody; PDCD 8 antibody; PDCD8 antibody; Programmed cell death 8 (apoptosis inducing factor) antibody; Programmed cell death 8 antibody; Programmed cell death 8 isoform 1 antibody; Programmed cell death 8 isoform 2 antibody; Programmed cell death 8 isoform 3 antibody; Programmed cell death protein 8 antibody; Programmed cell death protein 8 mitochondrial antibody; Programmed cell death protein 8 mitochondrial precursor antibody; Programmed cell death protein 8 mitochondrial precursor antibody; Striatal apoptosis inducing factor antibody

SEQUENCE SIMILARITIES

Belongs to the FAD-dependent oxidoreductase family.

TISSUE SPECIFICITY

Expressed in all tested tissues. Detected in muscle and skin fibroblasts (at protein level).; [Isoform 3]: Brain specific.; [Isoform 4]: Expressed in all tested tissues except brain.; [Isoform 5]: Isoform 5 is frequently down-regulated in human cancers.

POST-TRANSLATIONAL MODIFICATION

Under normal conditions, a 54-residue N-terminal segment is first proteolytically removed during or just after translocation into the mitochondrial intermembrane space (IMS) by the mitochondrial processing peptidase (MPP) to form the inner-membrane-anchored mature form (AIFmit). During apoptosis, it is further proteolytically processed at amino-acid position 101 leading to the generation of the mature form, which is confined to the mitochondrial IMS in a soluble form (AIFsol). AIFsol is released to the cytoplasm in response to specific death signals, and translocated to the nucleus, where it induces nuclear apoptosis in a caspase-independent manner.; Ubiquitination by XIAP/BIRC4 does not lead to proteasomal degradation. Ubiquitination at Lys-255 by XIAP/BIRC4 blocks its ability to bind DNA and induce chromatin degradation, thereby inhibiting its ability to induce cell death.

SUBCELLULAR LOCATION

Mitochondrion intermembrane space, Mitochondrion inner membrane, Cytoplasm, Nucleus.

FUNCTION

A key event in the apoptotic process is the opening of the mitochondrial permeability transition pore, an event that is regulated by Bcl-2 family proteins, resulting in the release of several proteins from the mitochondrial intermembrane space. Several of these proteins participate in apoptosis, including cytochrome c, procaspases 2, 3, and 9, and AIF (apoptosis-inducing factor). AIF has been shown to cause DNA fragmentation and chromatin condensation and to induce the release of cytochrome c and caspase-9 from mitochondria. Bcl-2 overexpression has been shown to prevent the release of AIF from mitochondria, but not to block its apoptogenic activity.

CITATIONS

  • Li, Xiaotong et al.

    Silkworm Pupa Protein Hydrolysate Induces Mitochondria-Dependent Apoptosis and S Phase Cell Cycle Arrest in Human Gastric Cancer SGC-7901 Cells. | International Journal of Molecular Sciences [2018]

  • Xie, Hongqing et al.

    Ethanolic extract of Cordyceps cicadae exerts antitumor effect on human gastric cancer SGC-7901 cells by inducing apoptosis, cell cycle arrest and endoplasmic reticulum stress. | Journal of Ethnopharmacology [2019]