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Antibodies

B7-H6 Recombinant Rabbit monoclonal Antibody

Product Sizes
100μl
£689.00
HA751381-100UL
About this Product
SKU:
HA751381
Application:
Western Blot, Flow Cytometry, Immunoprecipitation
Buffer:
PBS (pH7.4).
translate.label.attr.clone:
PSH10-89
Clonality:
Monoclonal
Conjugate:
Non-conjugated
Host:
Rabbit
Immunogen:
Recombinant protein within human B7-H6 aa 1-262.
Isotype:
IgG
Molecular Weight:
Predicted band size: 51 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. Avoid repeated freeze / thaw cycles.
Supplier:
Huabio
Type:
Antibodies: Monoclonal Antibody
Western blot analysis of B7-H6 on different lysates with Rabbit anti-B7-H6 antibody (HA751381) at 1/2;000 dilution. Lane 1: K-562 cell lysate Lane 2: HepG2 cell lysate Lane 3: MCF7 cell lysate (negative) Lane 4: HeLa cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 51 kDa Observed band size: 75 kDa Exposure time: 2 minutes 41 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 (HA751381) 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 K-562 (positive) and Daudi (negative) labeling B7-H6 with Rabbit anti-B7-H6 antibody (HA751381) 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-B7-H6 antibody (HA751381) 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.
Flow cytometric analysis of MCF7 (left; negative) and K-562 (right; positive) cells labeling B7-H6. Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (HA751381; 1/5;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).
B7-H6 was immunoprecipitated from 0.2 mg K-562 cell lysate with HA751381 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751381 at 1/1;000 dilution. Mouse Anti-Rabbit IgG kappa light chain secondary antibody (M1208-2) at 1/5;000 dilution was used for 1 hour at room temperature. Lane 1: K-562 cell lysate (input) Lane 2: HA751381 IP in K-562 cell lysate Lane 3: Rabbit IgG instead of HA751381 in K-562 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 3 minutes; ECL: K1801