Anti-Human HER2/neu Recombinant Antibody (HuA21)
CAT#: TAB-050CT
Recombinant monoclonal antibody to HER2/neu. This antibody intended for the prophylaxis and treatment of HER2-positive breast cancer.
Published Data
Gene Expression
Inhib
Figure 1 HuA21 suppressed the proliferation of breast cancer cells.
The inhibitory effects of HuA21, trastuzumab (Tra), or the combination on the growth of the: BT-474 (A); BT-474/HR (B); and SKBR3 (C) cells. The cells were incubated with HuA21 or Tra at the indicated concentrations for 72 h. Cell viability was measured by the the cell counting kit-8 (CCK-8) assay. The curve charts represent the cell viability of the: BT-474 (A); BT-474/HR (B); and SKBR3 cells (C). The experiments are performed in triplicate, and the data are representative of three separate experiments. # p < 0.05 vs. control; & p < 0.05 vs. HuA21-treated cells.
Li, R., Hu, S., Chang, Y., Zhang, Z., Zha, Z., Huang, H., ... & Wei, W. (2016). Development and characterization of a humanized anti-HER2 antibody HuA21 with potent anti-tumor properties in breast cancer cells. International journal of molecular sciences, 17(4), 563.
Cty
Figure 2 HuA21-mediated ADCC against HER2-positive breast cancer cells.
HuA21-mediated ADCC against HER2-positive breast cancer cells BT-474 cells were pre-incubated with the indicated concentrations of HuA21 or trastuzumab (Tra). The ADCC experiments were performed in triplicate, and the data are representative of three separate experiments.
Li, R., Hu, S., Chang, Y., Zhang, Z., Zha, Z., Huang, H., ... & Wei, W. (2016). Development and characterization of a humanized anti-HER2 antibody HuA21 with potent anti-tumor properties in breast cancer cells. International journal of molecular sciences, 17(4), 563.
IHC
Figure 3 HuA21 suppressed the migration of breast cancer cells.
The representative images and bar graph indicate the inhibitory effects of HuA21 (10 µg/mL), trastuzumab (Tra, 10 µg/mL), or HuA21 (10 µg/mL) + Tra (10 µg/mL) on the migration of SKBR3 breast cancer cells. The cells were treated with HuA21, Tra, or HuA21 + Tra for 24 h. Magnification: 100×. * p < 0.05 vs. control; & p < 0.05 vs. HuA21-treated cells. The experiments were performed in triplicate, and the data are representative of three separate experiments.
Li, R., Hu, S., Chang, Y., Zhang, Z., Zha, Z., Huang, H., ... & Wei, W. (2016). Development and characterization of a humanized anti-HER2 antibody HuA21 with potent anti-tumor properties in breast cancer cells. International journal of molecular sciences, 17(4), 563.
WB
Figure 4 HuA21 inhibited HER2 receptor-related signaling pathways.
Western blot and the representative picture showing the bands of EGFR, p-HER2, total HER2, total HER3, p-Akt, total Akt, p-Erk1/2, and total Erk1/2 expressed in the BT-474 treated with HuA21, trastuzumab (Tra), or HuA21 + Tra. β-Actin was used as the loading control. * p < 0.05 vs. control; & p < 0.05 vs. HuA21-treated cells. The experiments were performed in triplicate, and the data are representative of three separate experiments.
Li, R., Hu, S., Chang, Y., Zhang, Z., Zha, Z., Huang, H., ... & Wei, W. (2016). Development and characterization of a humanized anti-HER2 antibody HuA21 with potent anti-tumor properties in breast cancer cells. International journal of molecular sciences, 17(4), 563.
IF
Figure 5 HuA21 enabled internalization in HER2-positive cancer cells.
The BT-474cells were incubated with FITC-conjugated HuA21 or FITC-conjugated trastuzumab (Tra) for the indicated time periods, and then the presence of HuA21 or Tra was revealed by fluorescence microscope. Representative photomicrographs showing HuA21 or Tra (Green: FITC-conjugated HuA21 or Tra, scale bar = 50 µm).
Li, R., Hu, S., Chang, Y., Zhang, Z., Zha, Z., Huang, H., ... & Wei, W. (2016). Development and characterization of a humanized anti-HER2 antibody HuA21 with potent anti-tumor properties in breast cancer cells. International journal of molecular sciences, 17(4), 563.
IF
Figure 6 HuA21 enabled internalization in HER2-positive cancer cells.
The BT-474 cells were incubated with HuA21 or Tra for 4 h, FITC-conjugated secondary antibody was added and the signal was detected by confocal microscopy (Green: FITC-conjugated HuA21 or Tra; nuclei (blue) was stained by 4',6-diamidino-2-phenylindole (DAPI), scale bar = 50 µm)
Li, R., Hu, S., Chang, Y., Zhang, Z., Zha, Z., Huang, H., ... & Wei, W. (2016). Development and characterization of a humanized anti-HER2 antibody HuA21 with potent anti-tumor properties in breast cancer cells. International journal of molecular sciences, 17(4), 563.
FC
Figure 7 HuA21 enabled internalization in HER2-positive cancer cells.
The BT-474cells were incubated with Tra or HuA21 for the indicated periods. HuA21 or Tra on the cellsurface was labeled with a FITC-conjugated secondary antibody and analyzed by flow cytometry.
Li, R., Hu, S., Chang, Y., Zhang, Z., Zha, Z., Huang, H., ... & Wei, W. (2016). Development and characterization of a humanized anti-HER2 antibody HuA21 with potent anti-tumor properties in breast cancer cells. International journal of molecular sciences, 17(4), 563.
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Subcellular Location
Figure 1 IF staining of human cell line U-251 MG
Immunofluorescent staining of human cell line U-251 MG shows localization to plasma membrane.
* Image credit: Image credit: Human Protein Atlas v21.proteinatlas.org/images/1060/169_D12_1_selected.jpg
Subcellular Location
Figure 2 IF staining of human cell line A-431
Immunofluorescent staining of human cell line A-431 shows localization to plasma membrane.
* Image credit: Image credit: Human Protein Atlas v21.proteinatlas.org/images/1338/if_selected.jpg
Normal Tissue
Figure 3 High expression in esophagus
* Image credit: Image credit: Human Protein Atlas v21.proteinatlas.org/images/1383/4682_B_5_1_val_selected.jpg
Normal Tissue
Figure 4 IHC staining of human rectum
Immunohistochemical staining of human rectum shows weak cytoplasmic positivity in glandular cells.
* Image credit: Image credit: Human Protein Atlas v21.proteinatlas.org/images/43/ihc_selected.jpg
Normal Tissue
Figure 5 IHC staining of human urinary bladder
Immunohistochemical staining of human urinary bladder shows moderate cytoplasmic and membranous positivity in urothelial cells.
* Image credit: Image credit: Human Protein Atlas v21.proteinatlas.org/images/20416/ihc_selected.jpg
Normal Tissue
Figure 6 Kidney
Collecting ducts Staining: Low Intensity: Moderate Quantity: <25% Distal tubules Staining: Low Intensity: Moderate Quantity: <25%
* Image credit: Image credit: Human Protein Atlas v21.proteinatlas.org/images/43/1054_A_7_5.jpg
Normal Tissue
Figure 7 Breast
Glandular cells Staining: Medium Intensity: Moderate Quantity: 75%-25% Location: Cytoplasmic/ membranous Myoepithelial cells Staining: Medium Intensity: Moderate Quantity: 75%-25% Location: Cytoplasmic/ membranous
* Image credit: Image credit: Human Protein Atlas v21.proteinatlas.org/images/20416/45831_B_3_4.jpg
RNA Expression
Figure 8 RNA cell line category: Group enriched (OE19, SK-BR-3)
Cell lines ordered by descending RNA expression order
* Image credit: Image credit: Human Protein Atlas v21.proteinatlas.org/ENSG00000141736-ERBB2
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Specifications
- Host Species
- Human
- Type
- Humanized antibody
- Specificity
- Human
- Species Reactivity
- Human
- Clone
- HuA21
- Applications
- Inhib, Cty, IHC, WB, IF, FC, ELISA
- Related Disease
- HER2-positive breast cancer
Applications
- Application Notes
- The HER2/neu antibody has been reported in applications of Inhibition, Cytotoxicity, Immunohistochemistry, Western Blot, Immunofluorescence, Flow Cytometry, Enzyme-linked Immunosorbent Assay.
For Cytotoxicity: A standard lactate dehydrogenase (LDH) release assay was used to calculate the BT-474 or SKBR3 cells in the presence of the peripheral blood mononuclear cells (PBMC) isolated from fresh whole blood obtained from a healthy donor at an effector: target ratio of 80:1/well. In brief, aliquots of 0.1 mL of breast cancer target cells (2 × 10⁴ cells) were dispensed into round-bottomed tubes, and 0.1 mL of the effector cells was added to the target cells to give a desired effector-to-target (E:T) cell ratios. The indicated concentrations of Tra and HuA21 were added and were incubated for 5.5 h. The LDH release was evaluated by measuring absorbance at 490 nm on a Multiskan Ascent 96-well plate reader.
For Western Blot: The cells in each group were collected. The total protein was extracted and its concentration was measured using a bicinchoninic acid assay. The total protein (30 µg) was separated by a 10% sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis and was electrotransformed onto a polyvinylidene difluoride membrane. The membranes were blocked with 5% non-fat milk and were then incubated with a primary antibody at 4 °C overnight. An HRP-conjugated secondary antibody and an HRP-conjugated Actin were added to the membrane, which was incubated at 37 °C for 2 h. The immunoreactive proteins were visualized by enhanced chemiluminescence, and the signal intensity was detected and quantified by Image Quant LAS4000mini.
Target
- Alternative Names
- ERBB2; erb-b2 receptor tyrosine kinase 2; NEU; NGL; HER2; TKR1; CD340; HER-2; MLN 19; HER-2/neu; receptor tyrosine-protein kinase erbB-2; herstatin; p185erbB2; proto-oncogene Neu; c-erb B2/neu protein; proto-oncogene c-ErbB-2; metastatic lymph node gene 19 protein; human epidermal growth factor receptor 2; neuro/glioblastoma derived oncogene homolog; tyrosine kinase-type cell surface receptor HER2; neuroblastoma/glioblastoma derived oncogene homolog; v-erb-b2 avian erythroblastic leukemia viral oncoprotein 2; v-erb-b2 avian erythroblastic leukemia viral oncogene homolog 2; v-erb-b2 erythroblastic leukemia viral oncogene homolog 2, neuro/glioblastoma derived oncogene homolog;
- Gene ID
- 2064
- UniProt ID
- P04626
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Related Diseases
Bladder Cancer
Non-small Cell Lung Cancer
Related Signaling Pathways
ErbB Signaling Pathway
Downloadable Resources
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Product Notes
This is a product of Creative Biolabs' Hi-Affi™ recombinant antibody portfolio, which has several benefits including:
• Increased sensitivity
• Confirmed specificity
• High repeatability
• Excellent batch-to-batch consistency
• Sustainable supply
• Animal-free production
See more details about Hi-Affi™ recombinant antibody benefits.
Datasheet
MSDS
COA
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Protocol & Troubleshooting
We have outlined the assay protocols, covering reagents, solutions, procedures, and troubleshooting tips for common issues in order to better assist clients in conducting experiments with our products. View the full list of Protocol & Troubleshooting.
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