Human/Mouse/Rat AIF Antibody

Catalog # Availability Size / Price Qty
AF5824
AF5824-SP
AIF in Human Colon.
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Product Details
Citations (1)
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Human/Mouse/Rat AIF Antibody Summary

Species Reactivity
Human, Mouse, Rat
Specificity
Detects human, mouse, and rat AIF in Western blots.
Source
Polyclonal Sheep IgG
Purification
Antigen Affinity-purified
Immunogen
E. coli-derived recombinant human AIF
Glu121-Asp613
Accession # O95831
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.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
Simple Western
10 µg/mL
See below
Immunohistochemistry
5-15 µg/mL
See below
Knockout Validated
AIF is specifically detected in HEK293T human embryonic kidney parental cell line but is not detectable in AIF knockout HEK293T cell line.
 

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Immunohistochemistry AIF antibody in Human Colon by Immunohistochemistry (IHC-P). View Larger

AIF in Human Colon. AIF was detected in immersion fixed paraffin-embedded sections of human colon using Sheep Anti-Human/Mouse/Rat AIF Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5824) at 10 µg/mL overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

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

Detection of Human/Mouse/Rat AIF by Western Blot. Western blot shows lysates of Jurkat human acute T cell leukemia cell line, Hepa 1-6 mouse hepatoma cell line, and L6 rat myoblast cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human/Mouse/Rat AIF Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5824) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for AIF at approximately 65 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.

Simple Western Detection of Human AIF antibody by Simple Western<SUP>TM</SUP>. View Larger

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 10 µg/mL of Sheep Anti-Human/Mouse/Rat AIF Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5824) followed by 1:50 dilution ofHRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

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

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 1 µg/mL of Sheep Anti-Human/Mouse/Rat AIF Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5824) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). 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.

Reconstitution Calculator

Reconstitution Calculator

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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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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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.

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
Entrez Gene IDs
9131 (Human); 26926 (Mouse); 83833 (Rat)
Alternate Names
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

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Citation for Human/Mouse/Rat AIF Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

  1. CRKL, AIFM3, AIF, BCL2, and UBASH3A during Human Kidney Development
    Authors: Mirela Lozic, Luka Minarik, Anita Racetin, Natalija Filipovic, Mirna Saraga Babic, Katarina Vukojevic
    International Journal of Molecular Sciences

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