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APAF-1 Antibody

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Western Blot: APAF1 Antibody [NB100-56074] - Analysis of Apaf 1 using this antibody. Jurkat: human T-cell leukemia cell line lysate. RS11-846: human B-cell leukemia cell line lysate. Human thymus: tissue lysate.
Western Blot: APAF1 Antibody [NB100-56074] - Analysis of Apaf 1 using this antibody. Jurkat: human T-cell leukemia cell line lysate. RS11-846: human B-cell leukemia cell line lysate. Human thymus: tissue lysate.
Immunocytochemistry/ Immunofluorescence: APAF1 Antibody [NB100-56074] - Formalin-fixed, paraffin-embedded sections from a human brain tumor tissue microarray stained for Apaf 1 expression using NB100-56074 at 1:2000. A ...read more
Immunohistochemistry: APAF-1 Antibody [NB100-56074] - APAF1 Antibody [NB100-56074] - Formalin-fixed, paraffin-embedded tissue sections of human brain tumors stained for Apaf 1 expression using NB100-56074 at 1:2000. A. ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, GeSpecies Glossary
Applications WB, ICC/IF, IHC, IHC-Fr, IHC-P, IP
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated

Order Details

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APAF-1 Antibody Summary

Immunogen
Full-length recombinant human Apaf1 protein was used as immunogen.
Specificity
Apaf 1
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
APAF1
Purity
Unpurified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.
Learn about the Innovator's Reward

Applications/Dilutions

Dilutions
  • Western Blot 1:1000-1:2000
  • Immunocytochemistry/Immunofluorescence
  • Immunohistochemistry
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin 1:1000-1:5000
  • Immunoprecipitation 1:50-1:200
Control
APAF-1 Knockout HeLa Cell Lysate
Publications
Read Publication using
NB100-56074 in the following applications:

Packaging, Storage & Formulations

Storage
Store at -20C. Avoid freeze-thaw cycles.
Buffer
Neat whole antisera
Preservative
0.05% Sodium Azide
Purity
Unpurified

Alternate Names for APAF-1 Antibody

  • APAF1
  • APAF-1
  • APAF-1apoptotic protease activating factor 1
  • apoptotic peptidase activating factor 1
  • apoptotic peptidase activating factor
  • apoptotic protease activating factor
  • apoptotic protease-activating factor 1
  • CED4
  • DKFZp781B1145
  • KIAA0413

Background

Apaf 1 (apoptosis protease-activating factor-1) is a key regulator of the mitochondrial apoptotic pathway (reviewed in Campioni et al, 2005), being the central element of the multimeric apoptosome. The apoptosome consists of cytochrome c, procaspase-9 and seven Apaf 1 monomers, and is considered to be core apoptotic machinery that executes mitochondria-dependent apoptosis. Cytochrome c, normally compartmentalized in the mitochorndria, is released into the cytoplasm following apoptotic stimuli. Apaf 1 binds cytochrome c in the cytoplasm and in the presence of dATP/ATP forms the apoptosome. The apoptosome binds procaspase-9 and promotes its autocatalytic activation. Active caspase-9, in turn, activates downstream caspases including 3, 6, and 7 contributing to the proteolytic caspase activation cascade which leads to cell death. Apaf 1 is an approx. 130 kDa protein. Alternative Apaf 1 splicing isoforms of Apaf 1 have been identified in mammalian cells, each having either or both an 11 amino acid insertion in the N-terminus and an additional WD40 repeat (43 aa) in the C-terminus; please refer to AceView of NCBI (http://www.ncbi.nih.gov/IEB/Research/Acembly/) for detailed information. Additionally, Apaf 1 can be proteolytically processed by active caspase-3 during apoptosis resulting in an Apaf 1 cleavage fragment of ~84 kDa (Lauber et al. 2001).

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Immunohistochemistry-Paraffin: Caspase-9 Antibody [NB100-56118] - Analysis Human Brain sections stained for cleaved Caspase-9 expression using this antibody at 1:2000. A) Section from a patient 24 hr after head trauma. B) Control: section from a patient with no known neurological disease or head injury. Hematoxylin-eosin counterstain.
Immunohistochemistry-Paraffin: Caspase-9 Antibody [NB100-56118] - Analysis of Capase-9 in mouse transplanted lung tumor section using anti-Caspase-9 antibody. Image from verified customer review.
Immunohistochemistry-Paraffin: Caspase-9 Antibody [NB100-56118] - Analysis of Dog Brain sections stained for Active/Cleaved Caspase-9 expression at 1:2000. A and B. The pattern of Caspase-9 staining may vary between different types of neurons. Hematoxylin-eosin counterstain.
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(16 Publications)
Species: Hu, Mu, Rt, Ca, Ge
Applications: WB, ICC/IF, IHC, IHC-Fr, IHC-P, IP, Flow-IC
Cytochrome c
Antibody (7H8.2C12)
NB100-56503
Western Blot: Cytochrome c Antibody (7H8.2C12) [NB100-56503] - Analysis using the biotin conjugate of NB100-56503. Detection of Cytochrome C in 15 mg of HeLa cell lysate using NB100-55775 at 1:1000. A 15 kDa band is detected.
Immunocytochemistry/Immunofluorescence: Cytochrome c Antibody (7H8.2C12) [NB100-56503] - p73 was detected in immersion fixed Hela human cell line using NB100-56503 at 25 ug/ml for 3 hours at room temperature. Cells were stained using the NorthernLights(TM) 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Staining was observed in the cytoplasm and mitochondria.
Immunohistochemistry-Paraffin: Cytochrome c Antibody (7H8.2C12) [NB100-56503] - Cytochrome C was detected in immersion fixed paraffin-embedded sections of human heart using anti-human mouse monoclonal antibody (Catalog # NB100-56503) at 1:200 dilution overnight at 4C. Tissue was stained using the VisuCyte anti-mouse HRP polymer detection reagent (Catalog # VC001) with DAB chromogen (brown) and counterstained with hematoxylin (blue).Images may not be copied, printed or otherwise disseminated without express written permission of Novus Biologicals a bio-techne brand.
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Applications: WB, Simple Western, Flow, IB, ICC/IF, IHC, IHC-P, Flow-CS, Flow-IC
Caspase-3 Antibody
(31A1067) - (Pro and ...
NB100-56708
Western Blot: Caspase-3 Antibody (31A1067) - (Pro and Active) [NB100-56708] - Analysis using the Azide Free version of NB100-56708. Detection of Caspase-3 activation in HeLa cells. Cells were treated with 2mM staurosporine for different time periods. Caspase-3 activation is determined by cleavage of procaspase-3.
Western Blot: Caspase-3 Antibody (31A1067) - (Pro and Active) [NB100-56708] - Image of anti-Caspase 3 (31A1067). Whole cell protein from Jurkat cells treated with and without 2 uM staurosporine as indicated was separated on a 4-15% gel by SDS-PAGE, transferred to 0.2 um PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 5 ug/ml anti-Caspase 3 in 1% milk, and detected with an anti-mouse HRP secondary antibody using a Femto sensitivity chemiluminescence reagent. Note the detection of both pro-caspase 3 at 35 kDa and the cleaved active caspase 3 at 15-17 kDa.
Western Blot: Caspase-3 Antibody (31A1067) - (Pro and Active) [NB100-56708] - Analysis using the Azide Free version of NB100-56708. Detection of Caspase-3 in multiple human tissues. The tissues shown are A) brain, B) heart, C) intestine, D) kidney, E) liver, F) lung, G) muscle, H) stomach, I) spleen, J) ovary, and K) testis.
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(135 Publications)
Species: Hu, Mu, Rt, Po, Ch, ChHa, Ma
Applications: WB, Simple Western, EM, Flow, HEStain, IB, ICC/IF, IHC, IHC-Fr, IHC-P, CyTOF-ready, KO
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NB100-56098
Western Blot: Bcl-2 Antibody [NB100-56098] - Analysis of Bcl-2 in whole cell lysate from Daoy cells. Cells were transfected with (1) scrambled control siRNA or (2) Bcl-2 siRNA. Image from verified customer review.
Western Blot: Bcl-2 Antibody [NB100-56098] - Analysis of whole cell lysate (WCL) of Jurkat cells using anti-Bcl-2 antibody (NB100-56098) at 1:1000 dilution. HRP conjugated goat anti-rabbit IgG (H+L) cross adsorbed secondary antibody was used with ECL substrate for the detection of Bcl2 antibody bound to the blotted protein. This Bcl2 antibody detected a specific band at expected molecular weight marker position of Bcl2 protein (26kDa).
Immunohistochemistry-Paraffin: Bcl-2 Antibody [NB100-56098] - Analysis of a FFPE human breast carcinoma tissue section using 1:1000 dilution of Bcl-2 antibody (NB100-56098) on a Bond Rx autostainer (Leica Biosystems). The assay involved 20 minutes of heat induced antigen retrieval (HIER) with 10mM sodium citrate buffer (pH 6.0) and endogenous peroxidase quenching using peroxide block. The sections were incubated with primary antibody for 30 minutes. Bond Polymer Refine Detection (Leica Biosystems) and DAB were used for signal detection which followed counterstaining with hematoxylin. Whole slide scanning and capturing of representative images (20X) were performed using Aperio AT2 (Leica Biosystems). This antibody generated a diffused cytoplasmic staining of BCL2 in the cancer cells with relatively intense signal in their nuclear membranes. Staining was performed by Histowiz.
(2 Reviews)
(8 Publications)
Species: Hu, Mu, Rt, Ca
Applications: WB, IHC, IHC-P, IP
bcl-x Antibody
[Unconjugated]
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Western blot shows lysates of A431 human epithelial carcinoma cell line (5 x 104 cells, lane 1 and 2.5 x 104 cells, lane 2) and NIH‑3T3 mouse embryonic fibroblast cell line. PVDF membrane was probed with 0.3 µg/mL of Rabbit Anti-Human Bcl‑x Antigen Affinity-purified Polyclonal Antibody (Catalog # AF800) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Bcl‑x at approximately 26 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
Bcl-x was immunoprecipitated from lysates (7 x 105 cells) of A431 human epithelial carcinoma cell line following incubation with 3 µg Rabbit Anti-Human Bcl‑x Antigen Affinity-purified Polyclonal Antibody (Catalog # AF800) for 30‑60 minutes at 4 °C. Bcl-x-antibody complexes were absorbed using Protein A Immunoprecipitin (Life Technologies). Immuno­precipitated Bcl-x was detected by Western blot using 0.3 µg/mL Rabbit Anti-Human Bcl‑x Antigen Affinity-purified Polyclonal Antibody (Catalog # AF800). View our recommended buffer recipes for immunoprecipitation.
Simple Western lane view shows lysates of A431 human epithelial carcinoma cell line and NIH‑3T3 mouse embryonic fibroblast cell line, loaded at 0.2 mg/mL. A specific band was detected for Bcl‑x at approximately 34 kDa (as indicated) using 3 µg/mL of Rabbit Anti-Human/Mouse Bcl‑x Antigen Affinity-purified Polyclonal Antibody (Catalog # AF800). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
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Western Blot: Bax Antibody (6A7) [NBP1-28566] - Total cell lysates from NIH/3T3 cells were resolved by electrophoresis, transferred to PVDF membrane, and probled with Mouse Anti-Bax UNLB.
Immunohistochemistry-Frozen: Bax Antibody (6A7) [NBP1-28566] - Mouse retina with no pretreatment
Western Blot: Bax Antibody (6A7) [NBP1-28566] - Analysis of lysates from human metastatic pancreatic cancer cell line L3.6pl using Bax Antibody (clone 6A7; Lot # B168-P780E). This image was submitted via reviews by a varified end user.
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Applications: WB, ELISA, Flow, ICC/IF, IHC, IHC-Fr, IP
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NB100-56116
Western Blot: Caspase-8 Antibody - (active/cleaved) [NB100-56116] - Analysis of active/cleaved Caspase 8 in NRK whole cell lysate using anti-active/cleaved Caspase 8 antibody. Image from verified customer review.
Immunohistochemistry-Paraffin: Caspase-8 Antibody - (active/cleaved) [NB100-56116] - Sections from a brain tissue array stained for Caspase-8 expression using NB100-56116 at 1:2000. A. Normal brain stem (1) and cortex (2). B. Higher magnification of cortex (from A). C. Higher magnification of brain stem (from A). D. Normal cerebellum showing caspase-8 staining in the Purkinge cells.
Immunohistochemistry-Paraffin: Caspase-8 Antibody - (active/cleaved) [NB100-56116] - Sections from a brain tumor tissue array stained for Caspase-8 expression using NB100-56116 at 1:2000. A. Anaplastic glioma (Grade III, left) and Gemistocytoma (Grade II, right) cores showing negative and positive staining for Caspase-8, respectively. B. Higher magnification of the Gemistocytoma tumor (from A).
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Western Blot: p53 Antibody (PAb 240) [NB200-103] - Analysis of p53 in MCF7 and HeLa lystates. Image courtesy of anonymous customer product review.
Immunocytochemistry/Immunofluorescence: p53 Antibody (PAb 240) [NB200-103] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. The cells were incubated with anti-p53 (PAb 240) [NB200-103] at a 1:200 dilution overnight at 4C and detected with an anti-mouse DyLight 488 (Green) at a 1:500 dilution. Actin was detected with Phalloidin 568 (Red) at a 1:200 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.
Immunohistochemistry-Paraffin: p53 Antibody (PAb 240) [NB200-103] - p53 was detected in immersion fixed paraffin-embedded sections of human prostate cancer using anti-human mouse monoclonal antibody (Catalog # NB200-103) at 1:200 dilution overnight at 4C. Tissue was stained using the VisuCyte anti-mouse HRP polymer detection reagent (Catalog # VC001) with DAB chromogen (brown) and counterstained with hematoxylin (blue). Images may not be copied, printed or otherwise disseminated without express written permission of Novus Biologicals a bio-techne brand.
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[Unconjugated]
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Western blot shows lysates of Jurkat human acute T cell leukemia cell line, C2C12 mouse myoblast cell line, and PC‑12 rat adrenal pheochromocytoma cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse/Rat XIAP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8221) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for XIAP at approximately 56 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4.
Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line and C2C12 mouse myoblast cell line, loaded at 0.2 mg/mL. A specific band was detected for XIAP at approximately 59 kDa (as indicated) using 5 µg/mL of Goat Anti-Human/Mouse/Rat XIAP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8221) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the12-230 kDa separation system.
XIAP was detected in immersion fixed paraffin-embedded sections of human lymphoma using 5 µg/mL Goat Anti-Human/Mouse/Rat XIAP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8221) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
(13 Publications)
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FLASH Antibody
NB500-218
Western Blot: FLASH Antibody [NB500-218] - Analysis of FLASH in NIH3T3 cells using this antibody at 1:500 dilution.
Species: Hu, Mu
Applications: WB
SMAC/Diablo
Antibody
NB100-56311
Western Blot: SMAC/Diablo Antibody [NB100-56311] - Analysis of Smac (DIABLO) in HeLa lysate using Smac antibody at 2 ug/ml.
Immunohistochemistry: SMAC/Diablo Antibody [NB100-56311] - Analysis of human prostate using Smac antibody at 10 ug/ml.
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[Unconjugated]
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Recombinant Human TNF-alpha (Catalog # 210‑TA) induces cytotoxicity in the L-929 mouse fibroblast cell line in the presence of the metabolic inhibitor actinomycin D. The ED50 for this effect is 25‑100 pg/mL.
1 µg/lane of Recombinant Human TNF-alpha was resolved by SDS-PAGE with silver staining, under reducing (R) conditions, showing a band at 17 kDa.
(24 Reviews)
(602 Publications)
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BID Antibody
[Unconjugated]
AF860
Western blot shows Jurkat human acute T cell leukemia cell line treated with apoptosis inducer anti-Fas for the indicated times. PVDF membrane was probed with 1 µg/mL Goat Anti-Human/Mouse BID Antigen Affinity-purified Polyclonal Antibody (Catalog # AF860) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for BID at approximately 20 kDa (as indicated). For additional reference short (5 seconds, left panel) and long (60 seconds, right panel) exposures are shown. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line, loaded at 0.2 mg/mL. A specific band was detected for BID at approximately 29 kDa (as indicated) using 10 µg/mL of Goat Anti-Human/Mouse BID Antigen Affinity-purified Polyclonal Antibody (Catalog # AF860) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the12-230 kDa separation system.
Western blot shows lysates of SVEC4‑10 mouse vascular endothelial cell line and Jurkat human acute T cell leukemia cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse BID Antigen Affinity-purified Polyclonal Antibody (Catalog # AF860) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for BID at approximately 20 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
(4 Reviews)
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Applications: WB, Simple Western, IP
AIF Antibody
[Unconjugated]
AF1457
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 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 Northern­Lights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (yellow; Catalog # NL004) and counter­stained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
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.
(6 Publications)
Species: Hu, Mu, Rt
Applications: WB, Simple Western, ICC, KO
PARP Antibody
(194C1439) - Cleaved
NB100-56599
Western Blot: PARP Antibody (194C1439) - Cleaved [NB100-56599] - Analysis of cleaved PARP in staurosporine-treated Jurkat cells at various time points, antibody at 2 ug/mL. The band corresponding to cleaved PARP is only seen in the treated samples. Anti-mouse Ig HRP conjugate was used in this test.
Immunocytochemistry/Immunofluorescence: PARP Antibody (194C1439) - Cleaved [NB100-56599] - A431 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.5% Triton-X100. The cells were incubated with anti-PARP Antibody (194C1439) at 5 ug/ml overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.
Western Blot: PARP Antibody (194C1439) - Cleaved [NB100-56599] - Analysis using 293T lysates expressing PARP. Image from verified customer review.
(5 Publications)
Species: Hu, Mu, Rt(-)
Applications: WB, Flow, ICC/IF, Flow-IC
p21/CIP1/CDKN1A
Antibody (WA-1)
NBP2-29463
Immunohistochemistry-Paraffin: p21/CIP1/CDKN1A Antibody (WA-1 (HJ21)) [NBP2-29463] - Formalin-fixed, paraffin-embedded human bladder carcinoma stained with p21 Monoclonal Antibody (WA-1).
(13 Publications)
Species: Hu, Mu, Rt, Pm
Applications: WB, Flow, ICC/IF, IHC, IHC-Fr, IHC-P
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Publications for APAF-1 Antibody (NB100-56074)(1)

We have publications tested in 1 confirmed species: Human.

We have publications tested in 1 application: IHC-P.


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WB Video Protocol
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Control Lysate(s)

APAF-1 Knockout HeLa Cell Lysate

Secondary Antibodies

 

Isotype Controls

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Blogs on APAF-1.

Check out the latest blog posts on APAF-1.
Caspase 9 - an important apoptosis marker
Caspases are essential mediators of programmed cell death and are needed for both the induction of apoptosis as well as for aiding the degradation of cellular structures. Initiator caspases (such as Caspase-9) sense and respond to various signals i...  Read full blog post.
Heat Shock Proteins: An Overview
Heat Shock Proteins (HSPs) are a ubiquitous group of molecular chaperone proteins that have evolved unique mechanisms, within their host cells, to facilitate survival in hostile environments such as heat, oxidative (hypoxia), pH and cold. Under permis...  Read full blog post.
The Role of the Caspase 3 Antibody in Apoptosis Research
The caspases are a group of cysteine protease enzymes essential to apoptosis, inflammation and necrosis. Caspase 3 has been identified as one of the key mediators of apoptosis.Human caspases form three distinct groups: cytokine activators, which are...  Read full blog post.

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Bioinformatics

Gene Symbol APAF1
Entrez
Uniprot