HER2 Fab antibody

• Catalog Number: orb420301
• Category: Antibodies
• Description: HER2 Fab antibody
• Clonality: Recombinant
• Clone Number: Abz518
• Tested applications: ELISA, FC, WB
• Reactivity: Human
• Isotype: IgG Fab
• Concentration: 0.59 mg/ml
• Dilution range: ELISA: 1:10,000 to 1:100,000, FC: 1:100, WB: 1:2,000
• Form/Appearance: Liquid (sterile filtered)
• Purity: Humanized Recombinant HER2 Fab fragment was expressed in E. coli cells. The purity was estimated to be > 90% by SDS-PAGE analysis.
• Conjugation: Unconjugated
• UniProt ID: P04626
• NCBI: NP_001005862.1
• Storage: Store vial at 2-8° C. Dilute only prior to immediate use.
• Buffer/Preservatives: 0.01% (w/v) Sodium Azide
• Alternative names: CD340, cluster of differentiation 340, ERBB2 (huma
• Note: For research use only
• Application notes: Humanized Recombinant Anti-HER2 Fab fragment Antibody has been tested for use in Flow Cytometry, Western Blot, and ELISA. This antibody recognizes structured HER2 and will work in western blot when the protein has not been denatured with DTT or bMe. Although not tested, this antibody could be useful in in IHC and in in-vivo and other cellular assays. Specific conditions for reactivity should be optimized by the end user.
• Expiration Date: 12 months from date of receipt.

ELISA results show the sensitivity of Humanized Recombinant Anti-human HER2 Fab fragment Antibody tested against purified HER2 protein. Each well was coated in duplicate with 1.0 µg of HER2. The starting dilution of antibody was 5.0 µg/ml.

Flow Cytometry of Humanized Recombinant Anti-human HER2 Fab fragment Antibody. Primary Antibody: Recombinant Anti-HER2 Fab fragment (red line) recognizes HER2 on SK-OV-3 cells while using Anti-human kappa FITC secondary antibody at 0.2 µL with a 56.8% positive cells. Controls used: (black line) no primary antibody, no secondary antibody, 0.6%; (green line) no primary antibody, Anti-human kappa FITC secondary antibody at 0.2 µL, 1.0%.

SDS-PAGE of Humanized Recombinant Anti-human HER2 Fab fragment Antibody. Lane 1: Molecular Weight Ladder. Lane 2: reduced recombinant HER2 Fab fragment. Lane 3: non-reduced recombinant HER2 Fab fragment.

Western Blot of Humanized Recombinant Anti-human HER2 Fab fragment Antibody. Lane 1 and 8: Molecular Weight Marker. Lane 2 and 6: 0.02 µg HER2. Lane 3 and 7: 20 µg non-spiked SKOV-3 lysate. Lane 4 and 9: 20 µg SKOV-3 lysate spiked with 0.05 µg HER2. Lane 5 and 10: 20 µg SKOV-3 lysate spiked with 0.02 µg HER2. Primary Antibody: Anti-HER2 Fab incubated with lanes 1-5 only. Secondatry Antibody: Anti-F(ab)'2 HRP conjugate at 1:5000. Anti-HER2 Fab antibody observed in native HER2 and spiked HER2 in SKOV-3 cell lysates.

Western Blot of Humanized Recombinant Anti-human HER2 Fab fragment Antibody. Lane 1: Molecular Weight Ladder. Lane 2, 5, and 8: Empty. Lane 3, 6, and 9: HER2 reduced. Lane 4, 7, 10: HER2 non-reduced. Protein Load: Lane 3-4: 0.5 µg. Lane 6-7: 0.1 µg. Lane 9-10: 0.02 µg. Primary Antibody: anti-HER2 Fab at 1:2000 for 2 hours at 4°C. Secondary Antibody: Anti-human kappa FITC conjugate at 1:1000 for 2 hours at 4°C. Exposure time 60s.