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PRODUCT CODE: ET1602-39

Caspase-3 Recombinant Rabbit Monoclonal Antibody [SR03-01] (ET1602-39)

  • Recombinant

Applications

  • WB

  • ICC/IF

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

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All lanes: Western blot analysis of Caspase-3 with anti-Caspase-3 antibody [SR03-01] (ET1602-39) at 1:500 dilution.<br /> Lane 1: Wild-type Hela whole cell lysate (20 µg).<br /> Lane 2: Caspase-3 knockout Hela whole cell lysate (20 µg).<br /> <br /> ET1602-39 was shown to specifically react with Caspase-3 in wild-type Hela cells. No band was observed when Caspase-3 knockout samples were tested. Wild-type and Caspase-3 knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1602-39, 1/500) and Loading control antibody (Rabbit anti-β-actin, R1207-1, 1/1,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG-HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
  • All lanes: Western blot analysis of Caspase-3 with anti-Caspase-3 antibody [SR03-01] (ET1602-39) at 1:500 dilution.<br /> Lane 1: Wild-type Hela whole cell lysate (20 µg).<br /> Lane 2: Caspase-3 knockout Hela whole cell lysate (20 µg).<br /> <br /> ET1602-39 was shown to specifically react with Caspase-3 in wild-type Hela cells. No band was observed when Caspase-3 knockout samples were tested. Wild-type and Caspase-3 knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1602-39, 1/500) and Loading control antibody (Rabbit anti-β-actin, R1207-1, 1/1,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG-HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
  • Western blot analysis of Caspase-3 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1602-39, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br /> Positive control: <br /> Lane 1: Hela cell lysate<br /> Lane 2: Jurkat cell lysate<br /> Lane 2: 293 cell lysate
  • ICC staining of Caspase-3 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1602-39, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Caspase-3 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1602-39, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-Caspase-3 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1602-39, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
All lanes: Western blot analysis of Caspase-3 with anti-Caspase-3 antibody [SR03-01] (ET1602-39) at 1:500 dilution.
Lane 1: Wild-type Hela whole cell lysate (20 µg).
Lane 2: Caspase-3 knockout Hela whole cell lysate (20 µg).

ET1602-39 was shown to specifically react with Caspase-3 in wild-type Hela cells. No band was observed when Caspase-3 knockout samples were tested. Wild-type and Caspase-3 knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1602-39, 1/500) and Loading control antibody (Rabbit anti-β-actin, R1207-1, 1/1,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG-HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

Applications

  • WB

  • ICC/IF

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Caspase-3 Recombinant Rabbit Monoclonal Antibody [SR03-01] (ET1602-39)

Immunogen

Synthetic peptide within human caspase-3 aa 60-100.

Host

Rabbit

Positive Control

Hela cell lysate, Jurkat cell lysate, 293 cell lysate, Hela, human tonsil tissue, human spleen tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SR03-01

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

32 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:500-1:5,000

  • ICC/IF

  • 1:50-1:100

  • IHC-P

  • 1:50-1:200

  • FC:1:20-1:50

  • IP

  • Use at an assay dependent concentration.

TARGET

UNIPROT #

P42574

PROTEIN NAME

Caspase-3

SYNONYMS

A830040C14Rik antibody; Apopain antibody; CASP-3 antibody; CASP3 antibody; CASP3_HUMAN antibody; Casp3a antibody; Caspase 3 antibody; Caspase 3, apoptosis-related cysteine peptidase antibody; Caspase 3, apoptosis-related cysteine protease antibody; Caspase 3, apoptosis-related cysteine protease a antibody; Caspase-3 subunit p12 antibody; CC3 antibody; CPP-32 antibody; CPP32 antibody; CPP32B antibody; Cysteine protease CPP32 antibody; EC 3.4.22.56 antibody; LICE antibody; mldy antibody; OTTHUMP00000165052 antibody; OTTHUMP00000165053 antibody; OTTHUMP00000165054 antibody; PARP cleavage protease antibody; Procaspase3 antibody; Protein Yama antibody; SCA 1 antibody; SCA-1 antibody; SREBP cleavage activity 1 antibody; Yama antibody

SEQUENCE SIMILARITIES

Belongs to the peptidase C14A family.

TISSUE SPECIFICITY

Highly expressed in lung, spleen, heart, liver and kidney. Moderate levels in brain and skeletal muscle, and low in testis. Also found in many cell lines, highest expression in cells of the immune system.

POST-TRANSLATIONAL MODIFICATION

Cleavage by granzyme B, caspase-6, caspase-8 and caspase-10 generates the two active subunits. Additional processing of the propeptides is likely due to the autocatalytic activity of the activated protease. Active heterodimers between the small subunit of caspase-7 protease and the large subunit of caspase-3 also occur and vice versa.; S-nitrosylated on its catalytic site cysteine in unstimulated human cell lines and denitrosylated upon activation of the Fas apoptotic pathway, associated with an increase in intracellular caspase activity. Fas therefore activates caspase-3 not only by inducing the cleavage of the caspase zymogen to its active subunits, but also by stimulating the denitrosylation of its active site thiol.

SUBCELLULAR LOCATION

Cytoplasm

FUNCTION

Caspase-3, also known as apopain, SCA-1, Yama and CPP32, is an aspartate-specific cysteine protease that belongs to the ICE subfamily of caspases. Caspase-3 is expressed in cells as an inactive precursor from which the p17 and p11 subunits of the mature caspase-3 are proteolytically generated during apoptosis. The caspase-3 precursor is first cleaved at Asp175-Ser176 to produce the p11 subunit and the p20 peptide. Subsequently, the p20 peptide is cleaved at Asp28-Ser29 to generate the mature p17 subunit. The active caspase-3 enzyme is a heterodimer composed of two p17 and two p11 subunits. At the onset of apoptosis, caspase-3 proteolytically cleaves PARP at an Asp216-Gly217 bond. During the execution of the apoptotic cascade, activated caspase-3 releases SREBP from the membrane of the ER in a proteolytic reaction that is distinct from their normal sterol-dependent activation. Caspase-3 cleaves and activates SREBPs between the basic helix-loop-helix leucine zipper domain and the membrane attachment domain. Caspase-3 also cleaves and activates caspase-6, -7 and -9. The human caspase-3 gene encodes a cytoplasmic protein that is highly expressed in lung, spleen, heart, liver, kidney and cells of the immune system.

CITATIONS

  • Du, Changzheng et al.

    5-Fluorouracil targets histone acetyltransferases p300/CBP in the treatment of colorectal cancer. | Cancer Letters [2017]

  • Jian Li; Caiqiao Zhang

    Metformin Prevents Follicular Atresia in Aging Laying Chickens through Activation of PI3K/AKT and Calcium Signaling Pathways

  • Yuling Mi; Caiqiao Zhang

    The Attenuating Effect of the Intraovarian Bone Morphogenetic Protein 4 on Age-Related Endoplasmic Reticulum Stress in Chicken Follicular Cells

  • Deng, Y. L., Chen, S., Wang...

    Prescription of Sageretia hamosa Brongn Relieved Goiter through Promoted Apoptosis of Thyroid Cells via miR-511-3p and PTEN/PI3K/Akt Pathway