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HER2 Human Recombinant Antibody

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  • HER2 Human Recombinant Antibody
  • Capture ELISA of HER2: As validation of the antibody, 1ug/mL of HER2 antigen and control proteins (Fc and BSA) are adsorbed onto Maxisorp plates, blocked with PBS + 0.5% BSA (PB), and washed 3 times with PBS + 0.05% Tween-20 (PT); 0.2ug/mL of anti-HER2 hIgG1 antibody in PBS were incubated for 30 min shaking RT, washed 6 times with PT. Anti-human kappa-HRP (1:5000 in PB) was incubated for 30 min, shaking RT, washed 6 times with PT. Colorimetric HRP reagents allowed for absorbance readings at 450nM. Capture ELISA of HER2: As validation of the antibody, 1ug/mL of HER2 antigen and control proteins (Fc and BSA) are adsorbed onto Maxisorp plates, blocked with PBS + 0.5% BSA (PB), and washed 3 times with PBS + 0.05% Tween-20 (PT); 0.2ug/mL of anti-HER2 hIgG1 antibody in PBS were incubated for 30 min shaking RT, washed 6 times with PT. Anti-human kappa-HRP (1:5000 in PB) was incubated for 30 min, shaking RT, washed 6 times with PT. Colorimetric HRP reagents allowed for absorbance readings at 450nM.
  • Isoform-specificity of HER2:  1µg/ml of antigens (EGFR, HER2, HER3) and control proteins (Fc and BSA) are adsorbed onto Maxisorp plates, blocked with PBS + 0.5% BSA (PB), and washed 3 times with PBS + 0.05% Tween-20 (PT); 0.2µg/mL of anti-HER2 hIgG1 antibody in PBS were incubated for 30 min, shaking RT, washed 6 times with PT. Anti-human kappa-HRP (1:5000 in PB) was incubated for 30 min, shaking RT, washed 6 times with PT. Colorimetric HRP reagents allowed for absorbance readings at 450nM. Isoform-specificity of HER2: 1µg/ml of antigens (EGFR, HER2, HER3) and control proteins (Fc and BSA) are adsorbed onto Maxisorp plates, blocked with PBS + 0.5% BSA (PB), and washed 3 times with PBS + 0.05% Tween-20 (PT); 0.2µg/mL of anti-HER2 hIgG1 antibody in PBS were incubated for 30 min, shaking RT, washed 6 times with PT. Anti-human kappa-HRP (1:5000 in PB) was incubated for 30 min, shaking RT, washed 6 times with PT. Colorimetric HRP reagents allowed for absorbance readings at 450nM.
  • Flow Cytometry of HER2: MCF-7 and SKBR-3 cells, with low and high levels of surface expressed HER2, were collected using a mild EDTA solution and washed in PBS.  5x105 cells were incubated with 1ug of anti-HER2 schIgG1 antibody in 100 uL PBS + 0.1% BSA for 30 min on ice, then washed twice with cold PBS + 0.1 % BSA. Sample was then incubated with Goat anti-Human Fab-488 (Jackson ImmunoResearch, diluted with 1:1000 PBS + 0.1% BSA) for 15-30 min, washed twice with cold PBS + 0.1% BSA and fixed with 2% paraformaldehyde and filtered prior to reading on a FACS machine. Flow Cytometry of HER2: MCF-7 and SKBR-3 cells, with low and high levels of surface expressed HER2, were collected using a mild EDTA solution and washed in PBS. 5x105 cells were incubated with 1ug of anti-HER2 schIgG1 antibody in 100 uL PBS + 0.1% BSA for 30 min on ice, then washed twice with cold PBS + 0.1 % BSA. Sample was then incubated with Goat anti-Human Fab-488 (Jackson ImmunoResearch, diluted with 1:1000 PBS + 0.1% BSA) for 15-30 min, washed twice with cold PBS + 0.1% BSA and fixed with 2% paraformaldehyde and filtered prior to reading on a FACS machine.
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HER2 Human Recombinant Antibody

SKU:
MM-0315

USD 275.00 /100 µg

Clonality: 
Monoclonal
Host: Human; made using CCAB phage display library
Reactivity: Human
Application: Flow Cytometry

QTY:

OVERVIEW

Target: 
Human HER2
Target background: 
This member of the epidermal growth factor (EGF) receptor family of receptor tyrosine kinases has no ligand binding domain of its own and therefore cannot bind growth factors. However, it binds tightly to other ligand-bound EGF receptor family members to form a heterodimer, stabilizing ligand binding and enhancing kinase-mediated activation of downstream signalling pathways, such as those involving mitogen-activated protein kinase and phosphatidylinositol-3 kinase. Amplification and/or overexpression of this gene has been reported in numerous cancers, including breast and ovarian tumours. [provided by RefSeq, Jul 2008]
Target alias: 
CD340, ERBB2, HER2, NEU, NGL
Immunogen: 
ECD of recombinant HER2, aa: Thr23-Thr652
Specificity: This antibody recognizes the ECD of HER2 (aa: Thr23-Thr652) and does not recognize EGFR and HER3
Clone ID: 
CR - 015 (generated by CCAB)
Isotype: 
hIgG1; also available as mIgG1, scIgG1, schmIgG2a
Preservative: 
None
Format: 
Lyophilized protein A purified in PBS pH7.4
Recommend starting dilution: 
Reconstitute with deionized water. Optimal dilution has to be determined by the user.
Limitations: 
Research Use Only; Manufactured by CCAB under a license using Life Technologies proprietary cell lines.
Storage: 
Lyophilized antibodies can be kept at 4ºC for up to 3 months and should be kept at -20ºC for long-term storage (2 years). To avoid freeze-thaw cycles, reconstituted antibodies should be aliquoted before freezing for long-term (1 year) storage (-80ºC) or kept at 4ºC for short-term usage (2 months). For maximum recovery of product, centrifuge the original vial prior to removing the cap. Further dilutions can be made with the assay buffer. After the maximum long-term storage period (2 years lyophilized or 1 year reconstituted) antibodies should be tested in your assay with a standard sample to verify if you have noticed any decrease in their efficacy.