Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
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Species reactivity: | Human, Mouse, Rat |
Applications: | WB, IHC-P, IF-Cell |
Clonality: | Monoclonal |
Clone number: | JE59-31 |
Form: | Liquid |
Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. 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: 52 kDa |
Isotype: | IgG |
Immunogen: | Recombinant protein within Human Beclin 1 aa 351-450 / 450. |
Positive control: | Raji, MG-63 cell lysate, Mouse cerebellum tissue lysate, Rat cerebellum tissue lysate, human cerebellum tissue lysates, NIH/3T3 cell lysate, C6 cell lysate, human colon carcinoma tissue, mouse brain tissue. |
Subcellular location: | Cytoplasm, Golgi apparatus, trans-Golgi network membrane, Endosome membrane, Endoplasmic reticulum membrane, Mitochondrion membrane, Endosome, Cytoplasmic vesicle, autophagosome. |
Recommended Dilutions:
WB IHC-P IF-Cell |
1:500-1:1,000 1:200 1:250 |
Uniprot #: | SwissProt: Q14457 Human | O88597 Mouse | Q91XJ1 Rat |
Alternative names: | APG6 ATG 6 ATG6 ATG6 autophagy related 6 homolog Bcl-2-interacting protein beclin Beclin 1 (coiled coil moesin like BCL2 interacting protein) Beclin 1 autophagy related Beclin-1 Beclin1 BECN 1 Becn1 BECN1_HUMAN Coiled coil myosin like BCL2 interacting protein Coiled-coil myosin-like BCL2-interacting protein GT197 Protein GT197 VPS 30 VPS30 |
Fig1:
Immunocytochemistry analysis of Raji cells labeling Beclin 1 with Rabbit anti-Beclin 1 antibody (HA721216) at 1/250 dilution. Cells were fixed in 80% precooled methanol for 5 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-Beclin 1 antibody (HA721216) at 1/250 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 (M1305-2, 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. |
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Fig2:
Western blot analysis of Beclin 1 on different lysates with Rabbit anti-Beclin 1 antibody (HA721216) at 1/1,000 dilution. Lane 1: MG-63 cell lysate (20 µg/Lane) Lane 2: Mouse cerebellum tissue lysate (40 µg/Lane) Lane 3: Rat cerebellum tissue lysate (40 µg/Lane) Predicted band size: 52 kDa Observed band size: 52 kDa Exposure time: Lane 1: 3 minutes; Lane 2-3: 30 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 (HA721216) 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. |
Fig3:
Western blot analysis of Beclin 1 on human cerebellum tissue lysates with Rabbit anti-Beclin 1 antibody (HA721216) at 1/500 dilution. Lysates/proteins at 20 µg/Lane. Predicted band size: 52 kDa Observed band size: 52 kDa Exposure time: 1 minute; 10% 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 (HA721216) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature. |
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Fig4:
Western blot analysis of Beclin 1 on different lysates with Rabbit anti-Beclin 1 antibody (HA721216) at 1/500 dilution. Lane 1: MG-63 cell lysate Lane 2: NIH/3T3 cell lysate Lane 3: C6 cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 52 kDa Observed band size: 52 kDa Exposure time: 1 minute; 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 (HA721216) at 1/500 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. |
Fig5:
Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-Beclin 1 antibody (HA721216) 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 (HA721216) 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. |
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Fig6:
Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-Beclin 1 antibody (HA721216) 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 (HA721216) 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. |