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Antikörper

Cleaved PARP Recombinant Rabbit monoclonal Antibody

Produktgrößen
100μl
£689,00
HA750136-100UL
Über dieses Produkt
SKU:
HA750136
Anwendung:
Western Blot, Immunoprecipitation
Buffer:
PBS (pH7.4).
translate.label.attr.clone:
SU0314
Klonalität:
Monoclonal
Konjugat:
Non-conjugated
Host:
Rabbit
Immunogen:
Synthetic peptide within Human PARP1 aa 200-249 / 1;014.
Isotyp:
IgG
Molekulargewicht:
Predicted band size: 89 kDa
Physischer Zustand:
Liquid
Reinheit:
Protein A affinity purified.
Versandbedingungen:
Blue Ice
Lagerbedingungen:
Store at +4[o]C after thawing. Aliquot store at -20[o]C or -80[o]C. Avoid repeated freeze / thaw cycles.
Hersteller:
Huabio
Typ:
Antibodies: Monoclonal Antibody
Western blot analysis of Cleaved PARP on different lysates with Rabbit anti-Cleaved PARP antibody (HA750136) at 1/2;000 dilution. Lane 1: HeLa whole cell lysate Lane 2: HeLa treated with 1μM staurosporine for 3 hours whole cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 89 kDa Observed band size: 89 kDa Exposure time: 1 minute 9 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 (HA750136) 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.
Western blot analysis of Cleaved PARP on different lysates with Rabbit anti-Cleaved PARP antibody (HA750136) at 1/500 dilution. Lane 1: Hela-si NT cell lysate (10 µg/Lane) Lane 2: Hela-si Cleaved PARP cell lysate (10 µg/Lane) Predicted band size: 89 kDa Observed band size: 89 kDa Exposure time: 3 minutes; 4-20% SDS-PAGE gel. ET1608-10 was shown to specifically react with Cleaved PARP in Hela-si NT cells. No band was observed when Hela-si Cleaved PARP sample was tested. Hela-si NT and Hela-si Cleaved PARP samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1608-10; 1/500) and Loading control antibody (Rabbit anti-GAPDH; ET1601-4; 1/10;000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-rabbit IgG-HRP Secondary Antibody (HA1001) at 1:300;000 dilution was used for 1 hour at room temperature.
Immunocytochemistry analysis of HeLa cells treated with 1μM staurosporine for 180 minutes labeling Cleaved PARP with Rabbit anti-Cleaved PARP antibody (HA750136) at 1/250 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature; permeabilized with 0.1% Triton X-100 in PBS for 5 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-Cleaved PARP antibody (HA750136) at 1/250 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 (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.
Cleaved PARP was immunoprecipitated from 0.2 mg Daudi cell lysate with HA750136 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA750136 at 1/500 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: Daudi cell lysate (input) Lane 2: HA750136 IP in Daudi cell lysate Lane 3: Rabbit IgG instead of HA750136 in Daudi cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 12 seconds; ECL: K1801