HA601059

KRAS Mouse Monoclonal Antibody [A8E5]

cat.: HA601059

Product Type:	Mouse monoclonal IgG1, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P, FC
Clonality:	Monoclonal
Clone number:	A8E5

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 21 kDa
Isotype:	IgG1
Immunogen:	Recombinant protein within human KRAS aa 2-186.
Positive control:	Rat brain tissue lysate, PC-12 cell lysate, human kidney tissue lysate, Jurkat cell lysate, NIH/3T3, human lung carcinoma tissue, Hela.
Subcellular location:	Cell membrane. Cytoplasm.
Recommended Dilutions: WB IF-Cell IHC-P FC	1:1,000 1:100 1:1100 1:500-1:1,000
Uniprot #:	SwissProt: P01116 Human \| P32883 Mouse \| P08644 Rat
Alternative names:	c Ki ras2 c Kirsten ras protein c-K-ras c-Ki-ras Cellular c Ki ras2 proto oncogene Cellular transforming proto oncogene CFC2 cK Ras GTPase KRas K RAS p21 protein K RAS2A K RAS2B K RAS4A K RAS4B K-Ras 2 KI RAS Ki-Ras KIRSTEN MURINE SARCOMA VIRUS 2 Kirsten rat sarcoma 2 viral (v Ki ras2) oncogene homolog Kirsten rat sarcoma viral oncogene homolog KRAS KRAS proto oncogene, GTPase KRAS1 KRAS2 N-terminally processed NS NS3 Oncogene KRAS2 p21ras PR310 c K ras oncogene PR310 cK ras oncogene RALD RASK_HUMAN RASK2 Transforming protein p21 v Ki ras2 Kirsten rat sarcoma 2 viral oncogene homolog v Ki ras2 Kirsten rat sarcoma viral oncogene homolog

Images

	Fig1: Western blot analysis of KRAS on different lysates with Mouse anti-KRAS antibody (HA601059) at 1/1,000 dilution. Lane 1: Rat brain tissue lysate (20 µg/Lane) Lane 2: PC-12 cell lysate (10 µg/Lane) Lane 3: Human kidney tissue lysate (20 µg/Lane) Lane 4: Jurkat cell lysate (10 µg/Lane) Predicted band size: 21 kDa Observed band size: 21 kDa Exposure time: 2 minutes; 15% 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 (HA601059) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:100,000 dilution was used for 1 hour at room temperature.
	Fig2: Immunocytochemistry analysis of NIH/3T3 cells labeling KRAS with Mouse anti-KRAS antibody (HA601059) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 30 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 2% BSA for 30 minutes at room temperature. Cells were then incubated with Mouse anti-KRAS antibody (HA601059) at 1/100 dilution in 2% BSA overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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.
	Fig3: Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue with Mouse anti-KRAS antibody (HA601059) at 1/1,100 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 (HA601059) at 1/1,100 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.

Fig4: Flow cytometric analysis of Hela cells labeling KRAS.

Cells were fixed and permeabilized, and then blocked with 2% negative goat serum for 15 minutes at room temperature.Then stained with the primary antibody (HA601059, 1ug/ml) (red) compared with Mouse IgG1 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-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Fig5: Flow cytometric analysis of NIH/3T3 cells labeling KRAS.

Cells were fixed and permeabilized, and then blocked with 2% negative goat serum for 15 minutes at room temperature.Then stained with the primary antibody (HA601059, 1ug/ml) (red) compared with Mouse IgG1 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-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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