Ras Recombinant Rabbit Monoclonal Antibody [SA39-05]
cat.: ET1601-16
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat, Zebrafish
Applications: WB, IF-Cell, FC, IP
Clonality: Monoclonal
Clone number: SA39-05
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: 21 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human Ras aa 7-56 / 189.
Positive control: 293T cell lysate, MCF7 cell lysate, NIH/3T3 cell lysate, Neuro-2a cell lysate, PC-12 cell lysate, Mouse brain tissue lysate, MCF7, NIH/3T3, PC-12.
Subcellular location: Cytoplasm, Cell membrane, Golgi apparatus.
Recommended Dilutions:
  WB
  IF-Cell
  FC
  IP
1:1,000-1:2,000
1:100
1:1,000
1-2μg/sample
Uniprot #: SwissProt: P01111 Human | P01112 Human | P01116 Human | P08556 Mouse | P32883 Mouse | Q61411 Mouse | P08644 Rat | P20171 Rat | Q04970 Rat
Alternative names: C-BAS/HAS c-H-ras C-HA-RAS1 CTLO GTPase HRas GTPase KRas GTPase NRas H-Ras-1 H-RASIDX Ha-Ras HAMSV HRAS HRAS1 K RAS2A K RAS2B K RAS4A K RAS4B K-RAS KRAS KRAS1 KRAS2 N-RAS N-terminally processed NRAS NRAS1 p21ras RASH_HUMAN RASH1 RASK2 Transforming protein p21 v Ha ras Harvey rat sarcoma viral oncogene homolog v Ki ras2 Kirsten rat sarcoma viral oncogene homolog v ras neuroblastoma RAS viral oncogene homolog
Images
ET1601-16_1.jpg Fig1: Western blot analysis of Ras on different lysates with Rabbit anti-Ras antibody (ET1601-16) at 1/1,000 dilution.

Lane 1: 293T cell lysate (10 µg/Lane)
Lane 2: MCF7 cell lysate (10 µg/Lane)
Lane 3: NIH/3T3 cell lysate (10 µg/Lane)
Lane 4: Neuro-2a cell lysate (10 µg/Lane)
Lane 5: PC-12 cell lysate (10 µg/Lane)
Lane 6: Mouse brain tissue lysate (20 µg/Lane)

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

Exposure time: 30 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 (ET1601-16) 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.
ET1601-16_2.jpg Fig2: Immunocytochemistry analysis of MCF7 cells labeling Ras with Rabbit anti-Ras antibody (ET1601-16) 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-Ras antibody (ET1601-16) 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.
ET1601-16_3.jpg Fig3: Immunocytochemistry analysis of NIH/3T3 cells labeling Ras with Rabbit anti-Ras antibody (ET1601-16) 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-Ras antibody (ET1601-16) 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.
ET1601-16_4.jpg Fig4: Immunocytochemistry analysis of PC-12 cells labeling Ras with Rabbit anti-Ras antibody (ET1601-16) 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-Ras antibody (ET1601-16) 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.
ET1601-16_5.jpg Fig5: Flow cytometric analysis of MCF7 cells labeling Ras.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1601-16, 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).
ET1601-16_6.jpg Fig6: Flow cytometric analysis of NIH/3T3 cells labeling Ras.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1601-16, 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).
ET1601-16_7.jpg Fig7: Flow cytometric analysis of PC-12 cells labeling Ras.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1601-16, 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).
ET1601-16_8.jpg Fig8: Ras was immunoprecipitated from 0.2 mg MCF7 cell lysate with ET1601-16 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1601-16 at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: MCF7 cell lysate (input)
Lane 2: ET1601-16 IP in MCF7 cell lysate
Lane 3: Rabbit IgG instead of ET1601-16 in MCF7 cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 10 seconds; ECL: K1801
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.