Rab5 Recombinant Rabbit Monoclonal Antibody [ST47-04]
cat.: ET1609-27
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, IF-Cell
Clonality: Monoclonal
Clone number: ST47-04
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: 24 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human Rab5 aa 166-215 / 215.
Positive control: MCF-7 cell lysate, Hela cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, Neuro-2a cell lysate, PC-12 cell lysate, human colon cancer tissue, human prostate cancer tissue, PC-12.
Subcellular location: Cell membrane, Cytoplasm, Cell projection, Membrane, Endosome.
Recommended Dilutions:
  WB
  IHC-P
  IF-Cell

1:500-1:2,000
1:50-1:200
1:100
Uniprot #: SwissProt: P20339 Human | Q9CQD1 Mouse | M0RC99 Rat
Alternative names: RAB 5 RAB 5A RAB5A RAB5A member RAS oncogene family RAB5A_HUMAN RAS associated protein RAB5A Ras related protein Rab 5A Ras-related protein Rab-5A
Images
ET1609-27_1.jpg Fig1: Western blot analysis of Rab5 on different lysates with Rabbit anti-Rab5 antibody (ET1609-27) at 1/1,000 dilution.

Lane 1: MCF-7 cell lysate
Lane 2: Hela cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 24 kDa
Observed band size: 24 kDa

Exposure time: 1 minute;

12% 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 (ET1609-27) at 1/1,000 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.
ET1609-27_2.jpg Fig2: Western blot analysis of Rab5 on different lysates with Rabbit anti-Rab5 antibody (ET1609-27) at 1/1,000 dilution.

Lane 1: Mouse brain tissue lysate
Lane 2: Rat brain tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 24 kDa
Observed band size: 24 kDa

Exposure time: 2 minutes;

12% 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 (ET1609-27) at 1/1,000 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.
ET1609-27_3.jpg Fig3: Western blot analysis of Rab5 on different lysates with Rabbit anti-Rab5 antibody (ET1609-27) at 1/1,000 dilution.

Lane 1: Neuro-2a cell lysate
Lane 2: PC-12 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 24 kDa
Observed band size: 24 kDa

Exposure time: 3 minutes; ECL: K1801;

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 (ET1609-27) 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.
ET1609-27_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-Rab5 antibody (ET1609-27) 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 (ET1609-27) 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.
ET1609-27_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue with Rabbit anti-Rab5 antibody (ET1609-27) 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 (ET1609-27) 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.
ET1609-27_6.jpg Fig6: Immunocytochemistry analysis of PC-12 cells labeling Rab5 with Rabbit anti-Rab5 antibody (ET1609-27) at 1/100 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-Rab5 antibody (ET1609-27) at 1/100 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.