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AIFM1 Antibody (N-term)

Affinity Purified Rabbit Polyclonal Antibody (Pab)

Fluorescent image of U251 cells stained with AIFM1 (N-term) antibody. U251 cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min). Cells were then incubated with AP8910a AIFM1 (N-term) primary antibody (1:200, 2 h at room temperature). For secondary antibody, Alexa Fluor® 488 conjugated donkey anti-rabbit antibody (green) was used (1:1000, 1h). Cytoplasmic actin was counterstained with Alexa Fluor® 555 (red) conjugated Phalloidin (5.25 μM, 25 min). Pictures were taken on a Biorevo microscope (BZ-900, Keyence). Nuclei were counterstained with Hoechst 33342 (blue) (10 μg/ml, 5 min). AIFM1 (N-term) immunoreactivity is localized to the cytoplasm of U251 cells.
Fluorescent confocal image of U251 cells stained with AIFM1 (N-term) antibody. U251 cells were treated with Chloroquine (50 μM,16h), then fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min). Cells were then incubated with AP8910a AIFM1 (N-term) primary antibody (1:200, 2 h at room temperature). For secondary antibody, Alexa Fluor® 488 conjugated donkey anti-rabbit antibody (green) was used (1:1000, 1h). Nuclei were counterstained with Hoechst 33342 (blue) (10 μg/ml, 5 min). AIFM1 (N-term) immunoreactivity is localized to the cytoplasm of U251 cells.
Western blot analysis of AIFM1 Antibody (N-term) (Cat. #AP8910a) in 293 cell line lysates (35ug/lane). AIFM1 (arrow) was detected using the purified Pab.
Formalin-fixed and paraffin-embedded human brain tissue reacted with AIFM1 Antibody (N-term), which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated.
AIFM1 Antibody (N-term) (Cat. #AP8910a) flow cytometric analysis of 293 cells (right histogram) compared to a negative control cell (left histogram).FITC-conjugated goat-anti-rabbit secondary antibodies were used for the analysis.

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	IF, WB, IHC-P, FC, E
Primary Accession	O95831
Reactivity	Human
Host	Rabbit
Clonality	Polyclonal
Isotype	Rabbit IgG
Calculated MW	66901 Da
Antigen Region	70-98 aa

Additional Information
Gene ID	9131
Other Names	Apoptosis-inducing factor 1, mitochondrial, 111-, Programmed cell death protein 8, AIFM1, AIF, PDCD8
Target/Specificity	This AIFM1 antibody is generated from rabbits immunized with a KLH conjugated synthetic peptide between 70-98 amino acids from the N-terminal region of human AIFM1.
Dilution	IF~~1:200 WB~~1:1000 IHC-P~~1:100~500 FC~~1:10~50 E~~Use at an assay dependent concentration.
Format	Purified polyclonal antibody supplied in PBS with 0.09% (W/V) sodium azide. This antibody is purified through a protein A column, followed by peptide affinity purification.
Storage	Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
Precautions	AIFM1 Antibody (N-term) is for research use only and not for use in diagnostic or therapeutic procedures.

Protein Information
Name	AIFM1 (HGNC:8768)
Synonyms	AIF, PDCD8
Function	Functions both as NADH oxidoreductase and as regulator of apoptosis (PubMed:17094969, PubMed:20362274, PubMed:23217327, PubMed:33168626). In response to apoptotic stimuli, it is released from the mitochondrion intermembrane space into the cytosol and to the nucleus, where it functions as a proapoptotic factor in a caspase- independent pathway (PubMed:20362274). Release into the cytoplasm is mediated upon binding to poly-ADP-ribose chains (By similarity). The soluble form (AIFsol) found in the nucleus induces 'parthanatos' i.e. caspase-independent fragmentation of chromosomal DNA (PubMed:20362274). Binds to DNA in a sequence-independent manner (PubMed:27178839). Interacts with EIF3G, and thereby inhibits the EIF3 machinery and protein synthesis, and activates caspase-7 to amplify apoptosis (PubMed:17094969). Plays a critical role in caspase-independent, pyknotic cell death in hydrogen peroxide-exposed cells (PubMed:19418225). In contrast, participates in normal mitochondrial metabolism. Plays an important role in the regulation of respiratory chain biogenesis by interacting with CHCHD4 and controlling CHCHD4 mitochondrial import (PubMed:26004228).
Cellular Location	Mitochondrion intermembrane space. Mitochondrion inner membrane. Cytoplasm. Nucleus. Cytoplasm, perinuclear region. Note=Proteolytic cleavage during or just after translocation into the mitochondrial intermembrane space (IMS) results in the formation of an inner-membrane-anchored mature form (AIFmit). During apoptosis, further proteolytic processing leads to a 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 (PubMed:15775970). Release into the cytoplasm is mediated upon binding to poly-ADP-ribose chains (By similarity) Translocation into the nucleus is promoted by interaction with (auto- poly-ADP-ribosylated) processed form of PARP1 (PubMed:33168626) Colocalizes with EIF3G in the nucleus and perinuclear region (PubMed:17094969). {ECO:0000250\|UniProtKB:Q9Z0X1, ECO:0000269\|PubMed:15775970, ECO:0000269\|PubMed:17094969, ECO:0000269\|PubMed:33168626} [Isoform 4]: Mitochondrion. Cytoplasm, cytosol. Note=In pro-apoptotic conditions, is released from mitochondria to cytosol in a calpain/cathepsin-dependent manner.
Tissue Location	Expressed in all tested tissues (PubMed:16644725). Detected in muscle and skin fibroblasts (at protein level) (PubMed:23217327). Expressed in osteoblasts (at protein level) (PubMed:28842795). [Isoform 4]: Expressed in all tested tissues except brain.

Research Areas

For Research Use Only. Not For Use In Diagnostic Procedures.

Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol

BACKGROUND

AIFM1 is a flavoprotein essential for nuclear disassembly in apoptotic cells that is found in the mitochondrial intermembrane space in healthy cells. Induction of apoptosis results in the translocation of this protein to the nucleus where it effects chromosome condensation and fragmentation. In addition, this protein induces mitochondria to release the apoptogenic proteins cytochrome c and caspase-9.

REFERENCES

References for protein:
1.Daugas,E., et.al., FASEB J. 14 (5), 729-739 (2000)
2.Schulthess,F.T., et.al., PLoS ONE 4 (2), E4394 (2009)
References for U251 cell line:
1. Westermark B.; Pontén J.; Hugosson R. (1973).” Determinants for the establishment of permanent tissue culture lines from human gliomas”. Acta Pathol Microbiol Scand A. 81:791-805. [PMID: 4359449].
2. Pontén, J.,Westermark B. (1978).” Properties of Human Malignant Glioma Cells in Vitro”. Medical Biology 56: 184-193.[PMID: 359950].
3. Geng Y.;Kohli L.; Klocke B.J.; Roth K.A.(2010). “Chloroquine-induced autophagic vacuole accumulation and cell death in glioma cells is p53 independent”. Neuro Oncol. 12(5): 473–481.[ PMID: 20406898].

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¥ 1,250.00

Cat# AP8910a

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AIFM1 Antibody (N-term)

Affinity Purified Rabbit Polyclonal Antibody (Pab)

BACKGROUND

REFERENCES

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