RAB7 Recombinant Rabbit Monoclonal Antibody [SN202-03]
cat.: ET1611-96
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, FC
Clonality: Monoclonal
Clone number: SN202-03
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: 23 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human RAB7 aa 158-207 / 207.
Positive control: HeLa cell lysate, MCF7 cell lysate, Neuro-2a cell lysate, C2C12 cell lysate, PC-12 cell lysate, C6 cell lysate, Hela, SW480, NIH/3T3, human colon carcinoma tissue, human kidney tissue, mouse skeletal tissue, mouse kidney tissue, K562.
Subcellular location: Cytoplasmic vesicle, Lysosome membrane, Melanosome membrane, Lipid droplet.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
1:5,000
1:100-1:1,000
1:100-1:500
1:50-1:1,000
1:500-1:1,000
Uniprot #: SwissProt: P51149 Human | P51150 Mouse | P09527 Rat
Alternative names: CMT2B PRO2706 PSN RAB7, member RAS oncogene family RAB7A RAB7A, member RAS oncogene family RAB7A_HUMAN Ras associated protein RAB7 Ras related protein Rab7 Ras related protein Rab7a Ras-related protein Rab-7a
Images
ET1611-96_1.jpg Fig1: Western blot analysis of RAB7 on different lysates with Rabbit anti-RAB7 antibody (ET1611-96) at 1/5,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: MCF7 cell lysate
Lane 3: Neuro-2a cell lysate
Lane 4: C2C12 cell lysate
Lane 5: PC-12 cell lysate
Lane 6: C6 cell lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 23 kDa
Observed band size: 23 kDa

Exposure time: 17 seconds;

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 (ET1611-96) at 1/5,000 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.
ET1611-96_2.jpg Fig2: All lanes: Western blot analysis of RAB7 with anti-RAB7 antibody (ET1611-96) at 1:500 dilution.
Lane 1: Wild-type Hela whole cell lysate (10 µg).
Lane 2/3: RAB7 knockdown Hela whole cell lysate (10 µg).

ET1611-96 was shown to specifically react with RAB7 in wild-type Hela cells. Weakened bands were observed when RAB7 knockdown samples were tested. Wild-type and RAB7 knockdown 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 (ET1611-96, 1:500) was used in 5% BSA 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.
ET1611-96_3.png Fig3: Immunocytochemistry analysis of Hela cells labeling RAB7 with Rabbit anti-RAB7 antibody (ET1611-96) at 1/200 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-RAB7 antibody (ET1611-96) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. 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.
ET1611-96_4.jpg Fig4: Immunocytochemistry analysis of SW480 cells labeling RAB7 with Rabbit anti-RAB7 antibody (ET1611-96) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-RAB7 antibody (ET1611-96) at 1/50 dilution in 2% negative goat serum 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.
ET1611-96_5.png Fig5: Immunocytochemistry analysis of NIH/3T3 cells labeling RAB7 with Rabbit anti-RAB7 antibody (ET1611-96) at 1/200 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-RAB7 antibody (ET1611-96) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. 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.
ET1611-96_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-RAB7 antibody (ET1611-96) at 1/1,000 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 (ET1611-96) at 1/1,000 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.
ET1611-96_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-RAB7 antibody (ET1611-96) at 1/1,000 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 (ET1611-96) at 1/1,000 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.
ET1611-96_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded mouse skeletal tissue using anti-RAB7 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 (ET1611-96, 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.
ET1611-96_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue using anti-RAB7 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 (ET1611-96, 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.
ET1611-96_10.jpg Fig10: Flow cytometric analysis of RAB7 was done on K562 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1611-96, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
ET1611-96_11.jpg Fig11: Immunocytochemistry analysis of C6 cells labeling RAB7 with Rabbit anti-RAB7 antibody (ET1611-96) at 1/1,000 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS 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-RAB7 antibody (ET1611-96) at 1/1,000 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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