Actin Nucleation and Polymerization Antibody Sampler Kit
cat.: HAK21079
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.
Uniprot #: SwissProt: P60953 Human | P63000 Human | P60953 Human | P60766 Mouse | Q8CFN2 Rat | Q9Y6W5 Human | Q8BH43 Mouse | P07737 Human | P62962 Mouse | P62963 Rat | P61160 Human | P61161 Mouse | Q5M7U6 Rat | P61158 Human | Q99JY9 Mouse | Q4V7C7 Rat | O00401 Human | Q91YD9 Mouse
Entrez Gene: 313024 Rat
Images
HAK21079_1.jpg Fig1: Western blot analysis of RAC1 on different lysates with Rabbit anti-RAC1 antibody (HA722225) at 1/2,000 dilution.

Lane 1: HEK-293 cell lysate
Lane 2: Huh7 cell lysate
Lane 3: HUVEC cell lysate
Lane 4: HL-60 cell lysate
Lane 5: HeLa cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 21 kDa
Observed band size: 21 kDa

Exposure time: 2 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 (HA722225) at 1/2,000 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.
HAK21079_2.jpg Fig2: Immunocytochemistry analysis of HepG2 cells labeling CDC42 with Rabbit anti-CDC42 antibody (ET1701-7) at 1/500 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-CDC42 antibody (ET1701-7) at 1/500 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.
HAK21079_3.jpg Fig3: Western blot analysis of CDC42 on different lysates with Rabbit anti-CDC42 antibody (ET1701-7) at 1/1,000 dilution.

Lane 1: HepG2 cell lysate
Lane 2: MCF7 cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: C2C12 cell lysate
Lane 5: C6 cell lysate
Lane 6: Mouse brain tissue lysate
Lane 7: Rat brain tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 21 kDa
Observed band size: 21 kDa

Exposure time: 8 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 (ET1701-7) 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.
HAK21079_4.jpg Fig4: Immunocytochemistry analysis of NIH/3T3 cells labeling CDC42 with Rabbit anti-CDC42 antibody (ET1701-7) at 1/500 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-CDC42 antibody (ET1701-7) at 1/500 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.
HAK21079_5.jpg Fig5: Western blot analysis of Arp2 on different lysates with Rabbit anti-Arp2 antibody (HA721348) at 1/1,000 dilution.

Lane 1: Jurkat cell lysate
Lane 2: MCF7 cell lysate
Lane 3: SH-SY5Y cell lysate
Lane 4: HeLa cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 45 kDa
Observed band size: 40 kDa

Exposure time: 1 minute;

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 (HA721348) 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 1:200,000 dilution was used for 1 hour at room temperature.
HAK21079_6.jpg Fig6: Immunocytochemistry analysis of C6 cells labeling CDC42 with Rabbit anti-CDC42 antibody (ET1701-7) at 1/500 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-CDC42 antibody (ET1701-7) at 1/500 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.
HAK21079_7.jpg Fig7: Flow cytometric analysis of HepG2 cells labeling CDC42.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1701-7, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
HAK21079_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human heart tissue with Rabbit anti-Arp2 antibody (HA721348) 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 (HA721348) 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.
HAK21079_9.jpg Fig9: Immunocytochemistry analysis of SH-SY5Y cells labeling Arp2 with Rabbit anti-Arp2 antibody (HA721348) at 1/250 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-Arp2 antibody (HA721348) at 1/250 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.
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