Rabbit anti-Rat IgG F(ab')2 Secondary Antibody [HRP]

Western Blot: Rabbit F(ab')2 anti-Rat IgG Secondary Antibody [HRP] [NBP1-73682] - Lane 1: Rat IgG whole molecule. Lane 2: Rat IgG F(c) Fragment. Lane 3: Rat IgG Fab Fragment. Lane 4: Rat IgM Whole Molecule. All samples were reduced. Load: 50 ng per lane. Block: incubated with blocking buffer for 30 min at RT. Primary Antibody: Anti-Rat IgG F(ab')2 (RABBIT) Antibody 1:1,000 for 60 min at RT. Secondary Antibody: Anti-Rabbit IgG (GOAT) Peroxidase Conjugated Antibody 1:40,000 in incubated with blocking buffer for 30 min at RT. Predicted/Observed Size: 25 and 55 kDa for Rat IgG, 25 kDa for F(c) and Fab, 78 and 25 kDa for IgM. Rat F(c) migrates slightly higher.

Product Discontinued

Rabbit anti-Rat IgG F(ab')2 Secondary Antibody [HRP] Summary

• Immunogen: Rat IgG F(ab')2 fragment
• Specificity: No reaction was observed against Rat IgG F(c).
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • ELISA 1:10000 - 1:50000
  • Immunohistochemistry 1:500 - 1:2500
  • Western Blot 1:1000 - 1:10000
• Application Notes: This secondary antibody has been assayed against 1.0 ug of Rat IgG in a standard capture ELISA using ABTS (2,2'-azino-bis-[3-ethylbenthiazoline-6-sulfonic acid]) as a substrate for 30 minutes at room temperature. A working dilution of 1:500 to 1:3,000 of the reconstitution concentration is suggested for this product.
  
  NOTE: Do NOT add Sodium Azide (it inhibits HRP irreversibly).
• Readout System:
  • NovaLume Plus Chemiluminescent Reagent LUM-555

Packaging, Storage & Formulations

• Storage: Store lyophilized antibody at 4C in the dark. Aliquot reconstituted liquid and store at -20C. Avoid freeze-thaw cycles.
• Buffer: Lyophilized from 0.01 M Sodium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 10 mg/mL Bovine Serum Albumin (BSA) - Immunoglobulin and Protease free
• Preservative: 0.01% Gentamicin Sulfate
• Concentration: LYOPH
• Purity: Immunogen affinity purified
• Reconstitution Instructions: Reconstitute with 1.0 ml deionized water (or equivalent).

Notes

This product was prepared from monospecific antiserum by immunoaffinity chromatography using Rat IgG coupled to agarose beads. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Peroxidase, anti-Rabbit Serum, Rat IgG, Rat IgG F(ab')2 and Rat Serum

Store vial at 4C prior to restoration. For extended storage aliquot contents and freeze at -20C 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 4C as an undiluted liquid. Dilute only prior to immediate use.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Secondary Antibodies are guaranteed for 1 year from date of receipt.