Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat

Applications

Validated:

Knockout Validated, Western Blot, Immunocytochemistry, Simple Western

Cited:

Immunohistochemistry, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG
View all AIF Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human AIF
Glu121-Asp613
Accession # O95831

Specificity

Detects human, mouse, and rat mitochondria-processed AIF.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human/Mouse/Rat AIF Antibody

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

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.

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

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.

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

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.

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

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.

Detection of Human AIF antibody by Simple WesternTM.

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

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.

Applications for Human/Mouse/Rat AIF Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed Jurkat human acute T cell leukemia cell line treated with and without staurosporin, immersion fixed MCF-7 human breast cancer cell line, and immersion fixed HeLa human cervical epithelial carcinoma cell line

Knockout Validated

AIF is specifically detected in HEK293T human embryonic kidney parental cell line but is not detectable in AIF knockout HEK293T cell line.

Simple Western

1 µg/mL
Sample: Jurkat human acute T cell leukemia cell line and MCF‑7 human breast cancer cell line

Western Blot

0.1 µg/mL
Sample: 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

Reviewed Applications

Read 1 review rated 5 using AF1457 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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

Alternate Names

AIFM1, COXPD6, PDCD8

Entrez Gene IDs

9131 (Human); 26926 (Mouse); 83833 (Rat)

Gene Symbol

AIFM1

UniProt

O95831 (Human)

Additional AIF Products

Product Documents for Human/Mouse/Rat AIF Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human/Mouse/Rat AIF Antibody

For research use only

Related Research Areas

Bcl-2 Proteins and Regulators

Citations for Human/Mouse/Rat AIF Antibody

Customer Reviews for Human/Mouse/Rat AIF Antibody (1)

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  • Human/Mouse/Rat AIF Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: HepG2 human hepatocellular carcinoma cell line
    Species: Human
    Verified Customer | Posted 11/21/2022
    Cell lysate from HepG2 cells, 20 ug loaded. 10% gel, NIR western blot with LICOR secondary antibodies
    Human/Mouse/Rat AIF Antibody AF1457

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Protocols

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