Specificity |
GenScript Anti-Rabbit IgG Antibody (M205) recognizes the native rabbit IgG. This antibody may cross react with denatured IgG at high concentrations. The monoclonal antibody has no cross-reactivity with mouse, rat, bovine, goat, or sheep immunoglobulins and limited cross-reactivity with human immunoglobulins. |
Host Species |
Mouse |
Immunogen |
Rabbit IgG(H&L) |
Species Reactivity |
Rabbit |
Conjugate |
Horseradish peroxidase (HRP) |
Working concentrations for specific applications should be determined by the investigator. The appropriate concentration may be affected by primary antibody affinity, antigen concentration, the sensitivity of the method of detection, temperature, the length of the incubations, and other factors. The suitability of this antibody for applications other than those listed below has not been determined. The following concentration ranges are recommended starting points for this product.Optimal working dilutions must be determined by the end user. |
Application |
Recommended Usage |
Western Blot |
1:5,000 -1:20,000 |
ELISA |
1:5,000 -1:20,000 |
Immunohistochemistry (IHC) |
User-optimized |
|
Form |
Liquid |
Storage Buffer |
Supplied in PBS (pH 7.4), containing 1% BSA, 50% glycerol and 0.01% thimerosal. |
Concentration |
Not applicable |
Storage Instructions |
The lyophilized product remains stable for up to 1 year at -20 °C from the date of receipt. Upon reconstitution, ti can be stored for 2-3 weeks at 2-8 °C or for up to 12 months at -20 °C or below. Avoid repeated freezing and thawing cycles. |
Purification |
Protein A affinity column |
Isotype |
Mouse IgG1,κ |
Clonality |
Monoclonal |
Clone ID |
M205 |
Western blot of IP samples with A01827 or conventional secondary antibodies.
Loading:
Lane 1,3: Protein A beads (40 µl) + Glutamine Synthetase Antibody, pAb, Rabbit (10 µg) + Mouse brain lysate (200 µg)
Lane 2,4: Mouse brain lysate (20 µg)
Primary antibody:
Lane 1~4: Glutamine Synthetase Antibody, pAb, Rabbit
Secondary antibody:
Lane 1,2: Mouse Anti-Rabbit IgG Antibody (M205) [HRP], mAb (GenScript, A01827), 1:5,000
Lane 3,4: Anti-RABBIT IgG (H&L) (GOAT) Antibody Peroxidase Conjugated (Min X Bv Ch Gt GP Ham Hs Hu Ms Rt & Sh Serum Proteins), 1:5,000
Western blot of reduced and non-reduced rabbit IgG with A01827.
Loading:
Lane 1: Rabbit IgG (Reduced), 200 ng
Lane 2: Rabbit IgG (Non-reduced), 200 ng
We recommend loading no more than 200 ng of purified sample per lane
Detection antibody:
Lane 1~2: Mouse Anti-Rabbit IgG Antibody (M205) [HRP], mAb (GenScript, A01827), 1:5,000
Comparison of A01827 and Competitor Antibodies in Western Blot Analysis of IP Samples.
Loading:
Lanes 1,2,3: 40 µl Protein A beads + 10 µg Glutamine Synthetase Antibody, pAb, Rabbit + 200 µg Mouse brain lysate
Primary antibody:
Lanes 1,2,3: Glutamine Synthetase Antibody, pAb, Rabbit
Secondary antibody:
Lane 1: Mouse Anti-Rabbit IgG Antibody (M205) [HRP], mAb (GenScript, A01827), diluted 1:5,000
Lane 2: Competitor antibody (Conformation Specific), diluted 1:2,000
Lane 3: Conventional peroxidase-conjugated Goat Anti-RABBIT IgG (H&L) secondary antibody, diluted 1:5,000
Target Background |
Generally, goat or donkey anti-rabbit IgG polyclonal antibodies are used as secondary antibodies to detect rabbit primary antibodies. However, since polyclonal antibodies recognize many epitopes, the background and cross reactivity in many assays is usually high, making results hard to interpret. To minimize background, cross reactivity and improve the lot-to-lot consistency, a unique Anti-Rabbit IgG Antibody (M205) has been developed to use as a secondary antibody. Simply substitute conventional HRP or fluorescent reagents for unparalleled clarity in results and high quality results in immunoprecipitation, Western Blot, ELISA, Immunohistochemistry and Immunofluorescence. Anti-Rabbit IgG Antibody (M205) only recognizes native rabbit primary IgG antibodies. This antibody does not react with denatured or reduced rabbit IgG heavy or light chain. Immunoprecipitation and Western blots often suffer from primary antibody heavy and light chain contamination at approximately 50 and 25 kDa, which is caused by poor specificity of conventional polyclonal secondary antibodies derived from goat or other species. |
Synonyms |
Anti-Rabbit IgG [HRP] |
For laboratory research use only. Direct human use, including taking orally and injection and clinical use are forbidden.