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Antibodies

HA tag Recombinant Rabbit multiclonal Antibody

Product Sizes
100μl
£425.00
HA721750-100UL
About this Product
SKU:
HA721750
Application:
Western Blot, Immunoprecipitation
Buffer:
PBS (pH7.4); 0.05% BSA; 40% Glycerol. Preservative: 0.05% Sodium Azide.
translate.label.attr.clone:
PSH01-92
Conjugate:
Non-conjugated
Host:
Rabbit
Immunogen:
Synthetic peptide within N Terminal fusion protein and C Terminal fusion protein.
Isotype:
IgG
Physical State:
Liquid
Purity:
Protein A affinity purified.
Shipping Conditions:
Blue Ice
Storage Conditions:
Shipped at 4[o]C. Store at +4[o]C short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20[o]C long term.
Supplier:
Huabio
Type:
Antibodies
Western blot analysis of HA tag on different lysates with Rabbit anti-HA tag antibody (HA721750) at 1/5;000 dilution. Lane 1: 293T cell lysate Lane 2: 293T transfected with HA-tagged Nanos homolog 3 (N-terminal) cell lysate Lane 3: 293T transfected with HA-tagged LIPT1 (N-terminal) cell lysate Lane 4: 293T transfected with HA-tagged CXCL13 (C-terminal) cell lysate Lane 5: 293T transfected with HA-tagged CLK4 (C-terminal) cell lysate Lysates/proteins at 10 µg/Lane. Exposure time: 3 minutes; 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 (HA721750) at 1/5;000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50;000 dilution was used for 1 hour at room temperature.
Immunocytochemistry analysis of 293T cells labeling HA tag with Rabbit anti-HA tag antibody (HA721750) at 1/1;000 dilution. 293T cells; transfected with HA-tagged empty control; Nanos homolog 3 (N-terminal) or CLK4 (C-terminal) expression vector; respectively; were fixed in 4% paraformaldehyde for 10 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-HA tag antibody (HA721750) at 1/1;000 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.
HA tag was immunoprecipitated from 0.2 mg 293T transfected with HA-tagged Nanos homolog 3 (N-terminal) cell lysate with HA721750 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using M1008-1 at 1/20;000 dilution. Anti-Mouse IgG for IP Nano-secondary antibody (NBI02H) at 1/5;000 dilution was used for 1 hour at room temperature. Lane 1: 293T transfected with HA-tagged Nanos homolog 3 (N-terminal) cell lysate (input) Lane 2: HA721750 IP in 293T transfected with HA-tagged Nanos homolog 3 (N-terminal) cell lysate Lane 3: Mouse IgG instead of HA721750 in 293T transfected with HA-tagged Nanos homolog 3 (N-terminal) cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 35 seconds; ECL: K1801