Rabbit IgG (H&L) Secondary Antibody Peroxidase Conjugated Pre-adsorbed

SKU: orb347654

Description

RABBIT IgG (H&L) antibody (Peroxidase)

Images & Validation

• Tested Applications: DOT, ELISA, IHC, WB
• Dilution range: ELISA: 1:20,000 - 1:100,000, IHC: 1:1,000 - 1:5,000, WB: 1:2,000 - 1:10,000
• Reactivity: Rabbit
• Application Notes:
  Anti-Rabbit secondary antibody conjugated to horseradish peroxidase (HRP) generated in goat detects specifically rabbit IgG. This anti-rabbit HRP mx10 secondary antibody has been tested in ELISA, Dot Blot, and Western Blot. Although not tested, this product could be useful for Sandwich ELISA, titration assays, immunoprecipitation, immunohistochemistry as well as other HRP antibody based assays. Specific conditions for reactivity and signal detection should be optimized by the end user. 0.440 moles Peroxidase (HRP) per mole. Reconstitution Buffer: Restore with deionized water (or equivalent). Reconstitution Volume: 1.0 mL

Key Properties

• Antibody Type: Secondary Antibody
• Host: Goat
• Clonality: Polyclonal
• Isotype: IgG
• Immunogen: Rabbit IgG whole molecule
• Purity: Conjugated Second Antibody was prepared from monospecific antiserum by immunoaffinity chromatography using Rabbit IgG coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Peroxidase, anti-Goat Serum, Rabbit IgG and Rabbit Serum. No reaction was observed against Bovine, Chicken, Goat, Guinea Pig, Hamster, Horse, Human, Mouse, Rat and Sheep Serum Proteins.

Storage & Handling

• Storage: Store secondary antibody at 4° C prior to restoration. For extended storage aliquot contents and freeze at -20° C or below. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
• Form/Appearance: Lyophilized
• Buffer/Preservatives: 0.01% (w/v) Gentamicin Sulfate. Do NOT add Sodium Azide!
• Concentration: 2 mg/mL
• Disclaimer: For research use only

Alternative Names

goat anti-rabbit hrp, HRP conjugated secondary antibody, goat anti-rabbit secondary antibody, goat anti-rabbit IgG peroxidase conjugated secondary

ELISA Results of Goat Anti-Rabbit IgG Antibody Peroxidase Conjugated (Min X Bv, Ch, Gt, GP, Ham, Hs, Hu, Ms, Rt, Sh Serum Proteins) tested against purified Goat Anti-Rabbit IgG Antibody HRP Conjugated MX10. Each well was coated in duplicate with 10 µg of Rabbit IgG (p/n orb2652747) [Red Line], Bovine IgG (p/n orb2652780) [Green Line], Chicken IgG (p/n orb2652774) [Blue Line], Goat IgG (p/n orb2652767) [Purple Line], Guinea Pig, Hamster, Horse, Human, Mouse, Rat, and Sheep tested with similar results. The working dilution for Rabbit IgG is 1:109000. The starting dilution of antibody was 5 µg/ml and the X-axis represents the Log10 of a 3-fold dilution. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. Assay performed using 3% Fish Gel/PBS Blocking buffer, and TMB substrate (p/n orb348651).

HRP-conjugated Goat-Anti-Rabbit (p/n orb347654) secondary antibody was used at 1:40000 in orb348637 blocking buffer to detect a rabbit primary antibody by Western Blot. Anti-p27 antibody (orb344686, 1:1000 RT 30 minutes) showed detection of 0.1 µg of recombinant p27 protein. Lane 1: Molecular weight markers. Lane 2: MBP-p27 fusion protein (arrow; expected MW: 73.3 kDa). Lane 3: MBP alone. Protein was run on a 4-20% gel, then transferred to 0.45 µm nitrocellulose and blocked with 1% BSA-TTBS (p/n orb348540, diluted to 1X) overnight at 4°C.

Biorbyt secondary antibodies detect rabbit primary antibodies in a variety of platforms. Shown here is a serial 1:1 dilution of control rabbit IgG protein (orb2652747, 250 ng starting total load) co incubated with Biorbyt HRP conjugated Goat anti Rabbit IgG (orb347654) and Dylight 649 conjugated goat anti Rabbit 1:20K in orb348637.

Western Blot of Unconjugated Anti-Rabbit IgG (H&L) (GOAT) Antibody (Min X Bv, Ch, Gt, GP, Ham, Hs, Hu, Ms, Rt & Sh Serum Proteins) (p/n orb347649). Lane M: 3 µl Molecular Ladder. Lane 1: Rabbit IgG whole molecule (p/n orb2652747). Lane 2: Rabbit IgG F(ab) Fragment (p/n orb2652745). Lane 3: Rabbit IgG F(c) Fragment (p/n orb346280). Lane 4: Rabbit IgM Whole Molecule (p/n orb346313). Lane 5: Normal Rabbit Serum. All samples were reduced. Load: 50 ng per lane. Block: orb348637 for 30 min at RT. Primary Antibody: Anti-Rabbit IgG (H&L) (GOAT) Antibody (Min X Bv, Ch, Gt, GP, Ham, Hs, Hu, Ms, Rt & Sh Serum Proteins) (p/n orb347649) 1:1000 for 60 min at RT. Secondary antibody: Anti-Goat IgG (DONKEY) Peroxidase Conjugated Antibody 1:40000 in orb348637 for 30 min at RT. Predicted/Observed Size: 25 and 50 kDa for Rabbit IgG and Serum, 25 kDa for F(c) and F(ab), 70 and 23 kDa for IgM. Rabbit F(c) migrates slightly higher.