RAB45 Recombinant Rabbit Monoclonal Antibody [JE65-33]
cat.: HA721097
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P
Clonality: Monoclonal
Clone number: JE65-33
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
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: 83 kDa
Isotype: IgG
Immunogen: Synthetic peptide within human RAB45 aa 81-130/740.
Positive control: C6 cell lysate, PC-12 cell lysate, Mouse brain tissue lysate, Mouse spleen tissue lysate, Rat brain tissue lysate, Rat spleen tissue lysate, K562 cell lysate, A431 cell lysate, RAW264.7 cell lysate, NIH/3T3 cell lysate, PC-12, human spleen tissue, mouse spleen tissue, rat spleen tissue, human thyroid tissue, rat large intestine tissue, Hela.
Subcellular location: Perinuclear region.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
1:500-1:5,000
1:100
1:100-1:200
Uniprot #: SwissProt: Q8IZ41 Human | Q5RI75 Mouse
Entrez Gene: 298138 Rat
Alternative names: FLJ31614 RAB45 RAB45 member RAS oncogene family RAS and EF hand domain containing RAS and EF hand domain containing protein Ras related protein Rab 45
Images
HA721097_1.jpg Fig1: Western blot analysis of RAB45 on different lysates with Rabbit anti-RAB45 antibody (HA721097) at 1/5,000 dilution.

Lane 1: C6 cell lysate
Lane 2: PC-12 cell lysate
Lane 3: Mouse brain tissue lysate
Lane 4: Mouse spleen tissue lysate
Lane 5: Rat brain tissue lysate
Lane 6: Rat spleen tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 83 kDa
Observed band size: 83 kDa

Exposure time: 1 minute 55 seconds; 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 (HA721097) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA721097_2.jpg Fig2: Western blot analysis of RAB45 on different lysates with Rabbit anti-RAB45 antibody (HA721097) at 1/500 dilution.

Lane 1: K562 cell lysate
Lane 2: A431 cell lysate
Lane 3: RAW264.7 cell lysate
Lane 4: NIH/3T3 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 83 kDa
Observed band size: 83 kDa

Exposure time: 2 minutes;

8% 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 (HA721097) 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.
HA721097_3.jpg Fig3: Immunocytochemistry analysis of PC-12 cells labeling RAB45 with Rabbit anti-RAB45 antibody (HA721097) 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-RAB45 antibody (HA721097) 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.
HA721097_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-RAB45 antibody (HA721097) 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 (HA721097) 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.
HA721097_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-RAB45 antibody (HA721097) 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 (HA721097) 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.
HA721097_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-RAB45 antibody (HA721097) 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 (HA721097) 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.
HA721097_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human thyroid tissue with Rabbit anti-RAB45 antibody (HA721097) at 1/100 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 (HA721097) at 1/100 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.
HA721097_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded rat large intestine tissue with Rabbit anti-RAB45 antibody (HA721097) at 1/100 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 (HA721097) at 1/100 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”.