HRP Conjugated Anti-mouse IgG for IP Nano-secondary antibody
Antibody Type
Secondary Antibodies
Immunogen
Mouse IgG
Conjugation
HRP
Key Feature
Clonality
Polyclonal
Isotype
IgG
Host Species
Alpaca
Tested Applications
IPWB
WB:1:1000-1:10000
Species Reactivity
Mouse
Concentration
1mg/ml
Purification
Affinity purified
Target Information
Function
Anti-mouse IgG Nano-secondary antibody for IP (HRP) is based on Monovalent, recombinant single domain antibodies to mouse IgG coupled to HRP, and the Anti-mouse IgG for IP Nano-secondary antibody (HRP) detects the non-reduced form of mouse IgG (IgG1, IgG2a, IgG2b, IgG3) selectively, no reactivity with the reduced, SDS-denatured forms. When performing immunopreciptiation (IP) followed by western blotting, the denatured mouse IgG light and heavy chains of the primary antibody used for IP run at approximately 25 and 50 kD, respectively, on the subsequent western blot and can often obscure bands of proteins that have similar molecular weights. Anti-mouse IgG for IP Nano-secondary antibody (HRP) detects the non-reduced form of mouse IgG, no reactivity with the reduced, SDS-denatured forms. When a protien was immunoprecipitated by an antibody derived from mouse, you can still use primary antibody derived from mouse to detect this protien. Using Anti-mouse IgG for IP Nano-secondary antibody (HRP) to detect the primary antibody, only native antibody can be staining, not denatured heavy and light chains.
Application
WB
Immunoprecipitating IgG and DYKDDDDK Tag (FLAG) in recombinant protein with falg on N-terminal. 25ng of Immunoprecipitated protein incubated with primary antibody (1/1000) for over night at 4℃. For western blotting a IP Detection Reagent(1/1000) was used to confirm successful immunoprecipation. Lane1/2: Immunoprecipitating IgG in recombinant protein with flag on N-terminal Lane3: Immunoprecipitating DYKDDDDK Tag (FLAG) in recombinant protein with flag on N-terminal
IP
PCNA was immunoprecipitated from 0.2 mg HeLa cell lysate with PCNA Recombinant Mouse Monoclonal Antibody at 2µg/25µl agarose. Western blot was performed from the immunoprecipitate using PCNA Recombinant Mouse Monoclonal Antibody at 1/10,000 dilution. Anti-Mouse IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input) Lane 2: PCNA Recombinant Mouse Monoclonal Antibody IP in HeLa cell lysate Lane 3: Mouse IgG instead of PCNA Recombinant Mouse Monoclonal Antibody in HeLa cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 24 seconds; ECL: K1801
