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Antibodies

Phospho-Raf1 (S259) Recombinant Rabbit monoclonal Antibody

Product Sizes
100μl
£689.00
HA750304-100UL
About this Product
SKU:
HA750304
Application:
Western Blot, IHC-Paraffin, Flow Cytometry
Buffer:
PBS (pH7.4).
translate.label.attr.clone:
SD85-07
Clonality:
Monoclonal
Conjugate:
Non-conjugated
Host:
Rabbit
Immunogen:
Synthetic phospho-peptide corresponding to residues surrounding Ser259 of Human Raf1 aa 236-285 / 648.
Isotype:
IgG
Molecular Weight:
Predicted band size: 73 kDa
Physical State:
Liquid
Purity:
Protein A affinity purified.
Shipping Conditions:
Blue Ice
Storage Conditions:
Store at +4[o]C after thawing. Aliquot store at -20[o]C or -80[o]C. Avoid repeated freeze / thaw cycles.
Supplier:
Huabio
Type:
Antibodies: Monoclonal Antibody
Western blot analysis of Phospho-Raf1 (S259) on different lysates with Rabbit anti-Phospho-Raf1 (S259) antibody (HA750304) at 1/2;000 dilution. Lane 1: HEK-293 cell lysate Lane 2: HeLa cell lysate Lane 3: NIH/3T3 cell lysate Lane 4: C6 cell lysate Lane 5: HEK-293 cell lysate; the membrane treated with λpp for 1 hour Lysates/proteins at 20 µg/Lane. Predicted band size: 73 kDa Observed band size: 65 kDa Exposure time: 59 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 (HA750304) 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.
Immunocytochemistry analysis of HEK-293 cells labeling Phospho-Raf1 (S259) with Rabbit anti-Phospho-Raf1 (S259) antibody (HA750304) 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-Phospho-Raf1 (S259) antibody (HA750304) 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.
Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue with Rabbit anti-Phospho-Raf1 (S259) antibody (HA750304) 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 (HA750304) 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.
Immunohistochemical analysis of paraffin-embedded mouse skeletal muscle tissue with Rabbit anti-Phospho-Raf1 (S259) antibody (HA750304) 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 (HA750304) 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.