WB
Western blot analysis of HA tag on different lysates with Rabbit anti-HA tag antibody 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 at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.ICC/IF
Immunocytochemistry analysis of 293T cells labeling HA tag with Rabbit anti-HA tag antibody 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 at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.IP
HA tag was immunoprecipitated from 0.2 mg 293T transfected with HA-tagged Nanos homolog 3 (N-terminal) cell lysate with Rabbit anti-HA tag antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Rabbit anti-HA tag antibody at 1/20,000 dilution. Anti-Mouse IgG for IP Nano-secondary antibody 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: Rabbit anti-HA tag antibody IP in 293T transfected with HA-tagged Nanos homolog 3 (N-terminal) cell lysate, Lane 3: Mouse IgG instead of Rabbit anti-HA tag antibody in 293T transfected with HA-tagged Nanos homolog 3 (N-terminal) cell lysate Blocking/Dilution buffer: 5% NFDM/TBST. Exposure time: 35 second.| Product Name | HA tag Recombinant Rabbit Multiclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide within N Terminal fusion protein and C Terminal fusion protein. |
| Clonality | Polyclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ICC/IFIPWB |
| WB:1:5000 ICC/IF:1:1000 IP:1-2μg/sample |
|
| Species Reactivity | All |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | HA epitope tag antibody HA1 antibody HA2 antibody hemagglutinin antibody Hemagglutinin HA1 chain antibody Hemagglutinin HA2 chain antibody |
|---|---|
| Function | Human influenza hemagglutinin (HA) is a surface glycoprotein required for the infectivity of the human virus. The HA tag is derived from the HA molecule corresponding to amino acids 98-106 has been extensively used as a general epitope tag in expression vectors. Many recombinant proteins have been engineered to express the HA tag, which does not appear to interfere with the bioactivity or the biodistribution of the recombinant protein. This tag facilitates the detection, isolation, and purification of the proteins. |
WB
Western blot analysis of HA tag on different lysates with Rabbit anti-HA tag antibody 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 at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
ICC/IF
Immunocytochemistry analysis of 293T cells labeling HA tag with Rabbit anti-HA tag antibody 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 at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
IP
HA tag was immunoprecipitated from 0.2 mg 293T transfected with HA-tagged Nanos homolog 3 (N-terminal) cell lysate with Rabbit anti-HA tag antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Rabbit anti-HA tag antibody at 1/20,000 dilution. Anti-Mouse IgG for IP Nano-secondary antibody 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: Rabbit anti-HA tag antibody IP in 293T transfected with HA-tagged Nanos homolog 3 (N-terminal) cell lysate, Lane 3: Mouse IgG instead of Rabbit anti-HA tag antibody in 293T transfected with HA-tagged Nanos homolog 3 (N-terminal) cell lysate Blocking/Dilution buffer: 5% NFDM/TBST. Exposure time: 35 second.| Application Notes | WB:1:5000 ICC/IF:1:1000 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | PBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
