Rab Family Antibody Sampler Kit
cat.: HAK21152
Product Type: Antibody Sampler Kit
Species reactivity: Human
Applications: WB
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
Uniprot #: SwissProt: P20338 Human | P56371 Mouse | P05714 Rat | P20339 Human | Q9CQD1 Mouse | M0RC99 Rat | P51149 Human | P51150 Mouse | P09527 Rat | P51151 Human | Q9R0M6 Mouse | Q99P75 Rat | P62491 Human | P62492 Mouse | P62494 Rat | O00194 Human | Q99P58 Mouse | Q99P74 Rat
Images
HAK21152_1.jpg Fig1: Western blot analysis of Rab11A on different lysates with Rabbit anti-Rab11A antibody (HA721552) at 1/1,000 dilution.

Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: A549 cell lysate (20 µg/Lane)
Lane 3: SH-SY5Y cell lysate (20 µg/Lane)
Lane 4: Neuro-2a cell lysate (20 µg/Lane)
Lane 5: C6 cell lysate (20 µg/Lane)
Lane 6: Mouse testis tissue lysate (40 µg/Lane)
Lane 7: Rat testis tissue lysate (40 µg/Lane)
Lane 8: Rat brain tissue lysate (40 µg/Lane)

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

Exposure time: 2 minutes 37 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 (HA721552) 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 1:100,000 dilution was used for 1 hour at room temperature.
HAK21152_2.jpg Fig2: Western blot analysis of RAB27B on different lysates with Rabbit anti-RAB27B antibody (HA723666) at 1/50,000 dilution.

Lane 1: U-87 MG cell lysate (20 µg/Lane)
Lane 2: Mouse stomach tissue lysate (30 µg/Lane)
Lane 2: Mouse skeletal muscle tissue lysate (negative) (30 µg/Lane)
Lane 2: Rat colon tissue lysate (30 µg/Lane)

Predicted band size: 25 kDa
Observed band size: 25 kDa

Exposure time: 10 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 (HA723666) at 1/50,000 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.
HAK21152_3.jpg Fig3: 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.
HAK21152_4.jpg Fig4: Western blot analysis of Rab9 on different lysates with Rabbit anti-Rab9 antibody (ET1602-29) at 1/1,000 dilution.

Lane 1: HepG2 cell lysate
Lane 2: K-562 cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: Mouse lung tissue lysate
Lane 5: PC-12 cell lysate
Lane 6: Rat lung tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 23 kDa
Observed band size: 23 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 (ET1602-29) 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.
HAK21152_5.jpg Fig5: Immunocytochemistry analysis of Neuro-2a cells labeling Rab11A with Rabbit anti-Rab11A antibody (HA721552) 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-Rab11A antibody (HA721552) 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.
HAK21152_6.jpg Fig6: Immunocytochemistry analysis of NIH/3T3 cells labeling Rab9 with Rabbit anti-Rab9 antibody (ET1602-29) 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-Rab9 antibody (ET1602-29) 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.
HAK21152_7.jpg Fig7: 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.
HAK21152_8.jpg Fig8: 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.
HAK21152_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded rat stomach tissue with Rabbit anti-Rab9 antibody (ET1602-29) 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 (ET1602-29) 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.
HAK21152_10.jpg Fig10: Western blot analysis of Rab9 on different lysates with Rabbit anti-Rab9 antibody (ET1602-29) at 1/1,000 dilution.

Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-Rab9 KD cell lysate

Lysates/proteins at 10 µg/Lane.

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

Exposure time: 1 minute; 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 (ET1602-29) at 1/1,000 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.
HAK21152_11.jpg Fig11: 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).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.