Apaf-1 Recombinant Rabbit Monoclonal Antibody [SY22-02]
cat.: ET1607-12
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse
Applications: WB, IF-Cell, IF-Tissue, IHC-P
Clonality: Monoclonal
Clone number: SY22-02
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 142 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human Apaf-1 aa 1-50 / 1,248.
Positive control: HUVEC cell lysates, SW480 cell lysates, HeLa, human tonsil tissue, mouse colon tissue, mouse skin tissue, mouse heart tissue, human skin tissue.
Subcellular location: Cytoplasm.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
1:500-1:1,000
1:50-1:200
1:50-1:200
1:50-1:200
Uniprot #: SwissProt: O14727 Human | O88879 Mouse
Alternative names: APAF 1 Apaf-1 APAF_HUMAN Apaf1 Apoptotic peptidase activating factor 1 Apoptotic protease activating factor 1 Apoptotic protease activating factor Apoptotic protease-activating factor 1 CED 4 CED4 KIAA0413
Images
ET1607-12_1.jpg Fig1: Western blot analysis of Apaf-1 on different lysates with Rabbit anti-Apaf-1 antibody (ET1607-12) at 1/1,000 dilution.

Lane 1: A549-WT cell lysate
Lane 2: A549-KD Apaf-1 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 142 kDa
Observed band size: 142 kDa

Exposure time: 56 seconds; ECL: K1802;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1607-12) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1607-12_2.jpg Fig2: Immunocytochemistry analysis of HeLa cells labeling Apaf-1 with Rabbit anti-Apaf-1 antibody (ET1607-12) at 1/200 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Apaf-1 antibody (ET1607-12) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1607-12_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human skin tissue with Rabbit anti-Apaf-1 antibody (ET1607-12) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1607-12) at 1/200 dilution for 1 hour 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.
ET1607-12_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Apaf-1 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 (ET1607-12, 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.
ET1607-12_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse colon tissue using anti-Apaf-1 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 (ET1607-12, 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.
ET1607-12_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse skin tissue using anti-Apaf-1 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 (ET1607-12, 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.
ET1607-12_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded mouse heart tissue using anti-Apaf-1 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 (ET1607-12, 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.
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