Beclin 1 Recombinant Rabbit Monoclonal Antibody [JE59-31]
cat.: HA721216
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
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
Images
HA721216_1.jpg 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.
HA721216_2.jpg 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.
HA721216_3.jpg 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.
HA721216_4.jpg 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.
HA721216_5.jpg 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.
HA721216_6.jpg 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.