Anti-Klp2: rabbit polyclonal
This antibody is recommended for detection of human klp2, which is abundant in dividing cells. AKIN13 has been tested with extracts from other species, and klp2 was detected in mouse and rat cell extracts (Fig. 2). The following protocols have been tested with this antibody:
AKIN13 is provided as an affinity purified rabbit polyclonal antibody. The antibody was raised against the tail region of human kinesin Hklp2 (KIF15, KNSL7). Native Hklp2 has a predicted molecular weight of 160 kDa. Proteolytic breakdown products of Hklp2 (e.g. 55 kDa) may be be present in Western blots of some cell extracts. A HeLa cell extract is included as a positive control for antibody reactivity (see Fig. 1). AKIN13 (50 µg of protein) is supplied as a lyophilized white powder.
Figure 1. Western blot analysis of anti-Hklp2 antibody. Protein samples were separated by electrophoresis and transferred to PVDF membrane as described in the methods. AKIN13 antibody was diluted to 500 ng/ml (1:500) for Western blot analysis. Hklp2 was detected in 20 µg of HeLa cell extract (see arrow). Molecular weight markers are from Invitrogen.
Figure 2. Western blot of cell extracts probed with AKIN13 antibody. Chemiluminescence detection of Hklp2 in 20 µg HeLa cell extracts (lane 1, arrow; breakdown product at 55 kD shown as arrow head). Other lanes shown on the blot are mouse 3T3 (lane 2), rat NRK (lane 3), drosophila S2 (lane 4), and xenopus A6 (lane 5) cell extracts (50 µg each). (Lane 1 was from another region of the gel).
Figure 3. Immunoprecipitation of human Hklp2 from HeLa extracts using AKIN13. Western blot of HeLa cell lysate (Lane 1), IP supernatant after immunoprecipitation (Lane 2), Hklp2 immunoprecipitated with AKIN13 (160 kDa, see arrow) eluted from Protein A beads (lane 3), and control immunoprecipitation reaction with normal rabbit IgG (lane 4). Lanes 3 and 4 also show IgG heavy chain (55 kDa, arrow head). The Western blot was probed with 500 ng/ml (1:500 dilution) of AKIN13 antibody.
|Figure 4. Immunofluorescence images of Hela cells stained with AKIN13 antibody. Cells were grown to semi-confluency and fixed with methanol. Immunofluorescence staining of Hklp2 (red) in HeLa cells is shown using 1 µg/ml (1:250 dilution) of AKIN13 antibody. Primary antibody was detected with a 1:500 dilution of goat anti-rabbit rhodamine conjugated antibody (Cat. # RG05). DNA (blue) was stained with 100 nM DAPI in PBS. Photograph taken with a 100X objective lens.
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Question 1: What is the antigen that this antibody was raised against?
Answer 1: The antibody was raised against the tail region of human kinesin Hklp2 (also known as KIF15 and/or KNSL7).
Question 2: What species has this antibody been tested against?
Answer 2: This antibody is recommended for detection of human klp2, which is abundant in dividing cells. AKIN13 has been tested with extracts from other species, and klp2 was detected in mouse and rat cell/tissue extracts.
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