Antibody Type: Mouse Monoclonal

Application: WB, ELISA, FCM, ICC/IF, IP

Reactivity: other

Mouse IgG1

6T48

other

1. This antibody can be used at 0.5-1 μg/mL with the appropriate secondary reagents to detect HA-tagged protein (amino-terminus HA tag or carboxy-terminus HA tag).
2. Profile of anti-HA Tag reactivity on 293 transfected cells analyzed by flow cytometry. 293 cells transfected with HA-ARG1 (Figure A), HA-mFABP4 (Figure B), ARG1-HA (Figure C) and mFABP4-HA (Figure D) were fixed and permeabilized according to the manufacturer's manual and subsequently stained with Purified Mouse Anti-HA Tag antibody Or positive control antibody for 30 min on ice. Cells were washed twice and incubated with 1 μg of a FITC Goat Anti-Mouse Ig secondary antibody for 30 min on ice. Cells were washed twice and analyzed by flow cytometry.
3. Immunofluorescence staining of HA-Tag in 293 cells, transfected with HA-ARG1 (L) or mock-transfected (R). Cells were fixed with 4% PFA, permeabilzed with 1% Triton X-100 in PBS, blocked with 10% serum, and incubated with SBI Mouse anti-HA-tag monoclonal antibody at 37°C 1 hour. Then cells were stained with the Alexa Fluor® 488-conjugated Goat Anti-mouse IgG secondary antibody (green).
4. Immunofluorescence staining of HA-Tag in 293 cells, transfected with HA-ARG1 (Figure A), HA-mFABP4 (Figure B), ARG1-HA (Figure C) and mFABP4-HA (Figure D). Cells were fixed with 4% PFA, permeabilzed with 1% Triton X-100 in PBS, blocked with 10% serum, and incubated with Mouse anti-HA-Tag monoclonal antibody at 37°C 1 hour. Then cells were stained with the Alexa Fluor® 488-conjugated Goat Anti-mouse IgG secondary antibody (green) and counterstained with DAPI (blue).
5. Anti-HA Tag was immunoprecipitated using:
-Lane A: HA-ARG1-myc transfected 293 cell lysate (0.5mg)
-Lane B: myc-ARG1-HA transfected 293 cell lysate (0.5mg)
-2 μL anti-HA Tag mouse monoclonal antibody and 60 μg of Immunomagnetic beads Protein G.
-Primary antibody:
-Anti-HA Tag rabbit monoclonal antibody, at 1:100 dilution
-Secondary
-Goat Anti-Mouse IgG H&L (Dylight800) at 1/15000 dilution.
-Developed using the Odyssey technique.
-Performed under reducing conditions.
6. Anti-HA Tag mouse monoclonal antibody at 1:1000 dilution.
-Lane A: HA-GST (Recombinant protein)(30ng)
-Lane B: GST-HA (Recombinant protein)(10ng)
-Lane C: HA-ARG1-myc transfected 293 cell lysate (2ug)
-Lane D: HA-mFABP4-myc transfected 293 cell lysate (2ug)
-Lane E: myc-mFABP4-HA transfected 293 cell lysate (0.5ug)
-Lane F: myc-ARG1-HA transfected 293 cell lysate (2ug)
-Secondary
-Goat Anti-Mouse IgG H&L (Dylight800) at 1/15000 dilution.
-Developed using the Odyssey technique.
-Performed under reducing conditions.

WB,ELISA,FCM,ICC/IF,IP

WB: 1:2000-1:5000; ELISA: 1:2000-1:5000; ICC-IF: 1:200-1:500; FCM: 1:25-1:100; IP: 2-8 μL/mg of lysate

Monoclonal

Unconjugated

HRP

2

A synthetic peptide: HA-tag sequence (YPYDVPDYA).

other