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Knockout Tested Rabbit Recombinant Monoclonal APAF1 antibody. Carrier free. Suitable for IP, WB and reacts with Human, Mouse, Rat samples.

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Images

Immunoprecipitation - Anti-APAF1 antibody [EPR21112-102] - BSA and Azide free (AB254248), expandable thumbnail
  • Western blot - Anti-APAF1 antibody [EPR21112-102] - BSA and Azide free (AB254248), expandable thumbnail
  • Western blot - Anti-APAF1 antibody [EPR21112-102] - BSA and Azide free (AB254248), expandable thumbnail

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Constituents: PBS

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPFlow CytWBIHC-PICC/IF
Human
Tested
Not recommended
Tested
Not recommended
Not recommended
Mouse
Expected
Not recommended
Tested
Not recommended
Not recommended
Rat
Expected
Not recommended
Tested
Not recommended
Not recommended

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Expected
Expected

Species
Mouse, Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Mouse, Rat, Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/1000
Notes

-

Species
Mouse
Dilution info
1/1000
Notes

-

Species
Rat
Dilution info
1/1000
Notes

-

Not recommended
Not recommended

Species
Mouse, Rat, Human
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Mouse, Rat, Human
Dilution info
-
Notes

-

Target data

Function

Oligomeric Apaf-1 mediates the cytochrome c-dependent autocatalytic activation of pro-caspase-9 (Apaf-3), leading to the activation of caspase-3 and apoptosis. This activation requires ATP. Isoform 6 is less effective in inducing apoptosis.

Alternative names

KIAA0413, APAF1, Apoptotic protease-activating factor 1, APAF-1

Recommended products

Knockout Tested Rabbit Recombinant Monoclonal APAF1 antibody. Carrier free. Suitable for IP, WB and reacts with Human, Mouse, Rat samples.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free
Yes
Clone number
EPR21112-102
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze

Notes

ab254248 is the carrier-free version of Anti-APAF1 antibody [EPR21112-102] ab234436.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

APAF1 also known as apoptotic protease activating factor 1 plays a mechanical role in apoptosis by forming a complex with other proteins to trigger cell death. The APAF1 protein has a molecular weight of approximately 130 kDa. It is expressed in many tissues but occurs highly in the brain and heart. APAF1 interacts with cytochrome c and procaspase-9 to promote the activation of the initiator caspase cascade.

Biological function summary

APAF1 functions as a central component of the apoptosome a multi-protein complex important for programmed cell death. This process is essential for normal development and cellular homeostasis. In the apoptosome APAF1 recruits and activates procaspase-9 leading to the activation of caspase-3. This cascade assures that cells die in a controlled manner preventing damage to surrounding tissues.

Pathways

APAF1 serves an important function in the intrinsic pathway of apoptosis. It mediates cytochrome c release from mitochondria a step essential for apoptosis initiation. Another important pathway involving APAF1 is the apoptotic signaling pathway. Proteins such as Bcl-2 family members and p53 influence APAF1's role in these pathways by regulating cytochrome c release and apoptosome assembly.

Associated diseases and disorders

APAF1 is notably associated with neurodegenerative diseases and certain cancers. Dysregulation of APAF1 can lead to undesirable apoptosis contributing to neurodegenerative conditions like Parkinson’s disease. Conversely reduced APAF1 activity can result in tumor cell survival impacting cancer progression. Altered expression of APAF1 affects its interaction with proteins like Bcl-2 which can inhibit apoptosis and promote cancer cell growth.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

3 product images

  • Immunoprecipitation - Anti-APAF1 antibody [EPR21112-102] - BSA and Azide free (ab254248), expandable thumbnail

    Immunoprecipitation - Anti-APAF1 antibody [EPR21112-102] - BSA and Azide free (ab254248)

    APAF1 was immunoprecipitated from 0.35 mg HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate with Anti-APAF1 antibody [EPR21112-102] ab234436 at 1/30 dilution. Western blot was performed from the immunoprecipitate using Anti-APAF1 antibody [EPR21112-102] ab234436 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used at 1/5000 dilution.
    Lane 1: HEK-293T whole cell lysate 10 μg (Input).
    Lane 2: Anti-APAF1 antibody [EPR21112-102] ab234436 IP in HEK-293T whole cell lysate.
    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-APAF1 antibody [EPR21112-102] ab234436 in HEK-293T whole cell lysate.
    Blocking/Dilution buffer: 5% NFDM/TBST.
    Exposure time: 10 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-APAF1 antibody [EPR21112-102] ab234436).

    All lanes: Immunoprecipitation - Anti-APAF1 antibody [EPR21112-102] (Anti-APAF1 antibody [EPR21112-102] ab234436)

    Predicted band size: 141 kDa, 92 kDa

    Observed band size: 94 kDa

  • Western blot - Anti-APAF1 antibody [EPR21112-102] - BSA and Azide free (ab254248), expandable thumbnail

    Western blot - Anti-APAF1 antibody [EPR21112-102] - BSA and Azide free (ab254248)

    Anti-APAF1 antibody [EPR21112-102] ab234436 was shown to specifically react with APAF1 in wild-type HAP1 cells as signal was lost in APAF1 knockout cells. Wild-type and APAF1 knockout samples were subjected to SDS-PAGE. Anti-APAF1 antibody [EPR21112-102] ab234436 and Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging.

    Lanes 1,2 and 5 were developed using a higher sensitivity ECL substrate.

    Exposure times: Lanes 1-2: 136 secs; Lanes 3-4: 3 mins; Lane 5: 136 secs.

    Blocking/dilution buffer: 5% NFDM/TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-APAF1 antibody [EPR21112-102] ab234436).

    All lanes: Western blot - Anti-APAF1 antibody [EPR21112-102] - BSA and Azide free (ab254248) at 1/1000 dilution

    Lane 1: Wild-type HAP1 whole cell lysate at 20 µg

    Lane 2: APAF1 knockout HAP1 whole cell lysate at 20 µg

    Lane 3: HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen), whole cell lysate at 20 µg

    Lane 4: HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

    Lane 5: A20 (mouse reticulum sarcoma B lymphocyte), whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Predicted band size: 141 kDa

  • Western blot - Anti-APAF1 antibody [EPR21112-102] - BSA and Azide free (ab254248), expandable thumbnail

    Western blot - Anti-APAF1 antibody [EPR21112-102] - BSA and Azide free (ab254248)

    Western blot: Anti-APAF1 antibody [EPR21112-102] (Anti-APAF1 antibody [EPR21112-102] ab234436) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (Anti-Calnexin antibody [CANX/1543] ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-APAF1 antibody [EPR21112-102] ab234436 was shown to bind specifically to APAF1. A band was observed at 140 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in APAF1 knockout cell line. To generate this image, wild-type and APAF1 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

    All lanes: Western blot - Anti-APAF1 antibody [EPR21112-102] (Anti-APAF1 antibody [EPR21112-102] ab234436) at 1/1000 dilution

    Lane 1: Wild-type HCT 116 cell lysate at 20 µg

    Lane 2: APAF1 knockout HCT 116 cell lysate at 20 µg

    Lane 3: Wild-type HAP1 cell lysate at 20 µg

    Lane 4: APAF1 knockout HAP1 cell lysate at 20 µg

    Secondary

    All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

    Performed under reducing conditions.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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