Anti-Human EGFR Recombinant Antibody (Panitumumab)

Name: Anti-Human EGFR Recombinant Antibody (Panitumumab)
SKU: TAB-020
Rating: 5 (1 reviews)

CAT#: TAB-020

Recombinant monoclonal antibody to Human EGFR. Panitumumab (INN), formerly ABX-EGF, is a fully human monoclonal antibody specific to the epidermal growth factor receptor (also known as EGF receptor, EGFR, ErbB-1 and HER1 in humans).

Published Data Tested Data Gene Expression

Figure 1 EGFR G465R mutant reveals almost complete abrogation of panitumumab and cetuximab binding.

EGFR mutation G465R abrogates binding of panitumumab and cetuximab at the protein level. Wile-type and mutant EGFR-Fc proteins were expressed and the binding of therapeutic antibodies to immobilized proteins was assessed by ELISA. Data are means from 3 experiments +/- SEM. * p<0.05, ** p<0.01, n.s. = not significant (student's T-test comparing binding of respective antibody to mutant versus wt EGFR-Fc).

Braig, F., März, M., Schieferdecker, A., Schulte, A., Voigt, M., Stein, A., ... & Engel, E. (2015). Epidermal growth factor receptor mutation mediates cross-resistance to panitumumab and cetuximab in gastrointestinal cancer. Oncotarget, 6(14), 12035.

Figure 2 EGFR G465R mutant reveals almost complete abrogation of panitumumab and cetuximab binding.

Panitumumab and cetuximab binding is abrogated in CHO cells transfected with EGFR G465R . EGFR negative CHO cells were transfected with wild type EGFR or mutants thereof. Binding of panitumumab, cetuximab or a control polyclonal EGFR antibody was assessed by FACS analysis 48h after transfection. FSC = forward scatter.

Figure 3 EGFR G465R mutation induces cross-resistance to panitumumab and cetuximab in an EGF-dependent Ba/ F3 cellular model.

A: EGFR signaling in EGF-dependent Ba/F3 model. Wt and S492R or G465R mutant EGFR-expressing Ba/F3 cells were cultured in the presence or absence of EGF and with addition of cetuximab, panitumumab, rituximab or erlotinib. After 2 hours, cells were harvested and EGFR/pEGFR expression analyzed by western blot analysis. B: Sensitivity of EGFR wt or EGFR G465R mutant- transfected Ba/F3 cells to treatment with EGFR-targeted antibodies. Ba/F3 cells were transformed to IL-3 independence with EGFR wt or mutant constructs and subsequently cultured in the presence or absence of EGF or with EGF in combination with panitumumab, cetuximab or control antibody rituximab. The number of viable cells was determined by trypan blue exclusion every 12 hours beginning 24 hours after seeding and plotted. Data are means from triplicate experiments +/-SEM.

Figure 4 In vitro EGFR targeting of panitumumab.

Immunofluorescence staining of UM-SCC-22B tumor cells and tissues for EGFR using panitumumab as the primary antibody.

Liu, Z., Liu, Y., Jia, B., Zhao, H., Jin, X., Li, F., ... & Wang, F. (2010). Epidermal growth factor receptor–targeted radioimmunotherapy of human head and neck cancer xenografts using 90Y-labeled fully human antibody panitumumab. Molecular cancer therapeutics, 1535-7163.

Figure 5 In vitro EGFR targeting of panitumumab.

In vitro inhibition of 125I-panitumumab binding to EGFR on UM-SCC-22B human head and neck cancer cells by DOTA-panitumumab and panitumumab. Points, mean (n = 3); bars, SD.

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Figure 1 Anti-Human EGFR Recombinant Antibody (TAB-020) in WB

Western blot analysis of TAB-020 was performed with Human EGFR protein.

TAB-020 incubation concentration: 2 ng/μL.
The secondary antibody: HRP-IgG (H+L)
Lane 1: Reducing Antigen (0.1 μg)
Lane 2: Reducing Antigen (0.3 μg)
Lane 3: Reducing Antigen (0.6 μg)

ELISA

Figure 2 Anti-Human EGFR Recombinant Antibody (TAB-020) in ELISA

ELISA analysis of TAB-020 was performed by coating with Human EGFR protein.

The secondary antibody: HRP-anti-Human IgG

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Subcellular Location and Protein Expression Figure 1 IF staining of human cell line A-431

Immunofluorescent staining of human cell line A-431 shows localization to plasma membrane & cell junctions.

* Image credit: Image credit: Human Protein Atlas https://v21.proteinatlas.org/images/18530/144_E9_2_selected.jpg

Subcellular Location and Protein Expression Figure 2 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 https://v21.proteinatlas.org/images/18530/143_E9_1_blue_red_green.jpg

Normal Tissue Figure 3 Colon

Glandular cells
Staining:Medium
Intensity: Moderate
Quantity:>75%
Location: Cytoplasmic/membranous

* Image credit: Image credit: Human Protein Atlas https://v21.proteinatlas.org/images/18530/41191_A_9_3.jpg

Normal Tissue Figure 4 Liver

Cholangiocytes
Staining:Medium
Intensity: Moderate
Quantity: 75%-25%
Location: Cytoplasmic/membranous

* Image credit: Image credit: Human Protein Atlas https://v21.proteinatlas.org/images/18530/41191_A_7_4.jpg

Normal Tissue Figure 5 Kidney

Bowman's capsule
Staining:Medium
Intensity: Moderate
Quantity: 75%-25%
Collecting ducts
Staining:Medium
Intensity: Strong
Quantity: <25%
Distal tubules
Staining:Medium
Intensity: Strong
Quantity: <25%
Proximal tubules (cell body)
Staining:Medium
Intensity: Strong
Quantity: <25%

* Image credit: Image credit: Human Protein Atlas https://v21.proteinatlas.org/images/18530/41191_A_9_5.jpg

Normal Tissue Figure 6 Testis

Leydig cells
Staining:Medium
Intensity: Moderate
Quantity:>75%
Pachytene spermatocytes
Staining:Medium
Intensity: Moderate
Quantity: 75%-25%
Round or early spermatids
Staining:Medium
Intensity: Moderate
Quantity: 75%-25%

* Image credit: Image credit: Human Protein Atlas https://v21.proteinatlas.org/images/18530/41191_A_6_6.jpg

Normal Tissue Figure 7 Placenta

Cytotrophoblasts
Staining:High
Intensity: Strong
Quantity:>75%
Decidual cells
Staining:Medium
Intensity: Moderate
Quantity:>75%
Hofbauer cells
Staining:High
Intensity: Strong
Quantity:>75%
Syncytiotrophoblasts - cell body
Staining:High
Intensity: Strong
Quantity:>75%
Syncytiotrophoblasts - microvilli
Staining:High
Intensity: Strong

* Image credit: Image credit: Human Protein Atlas https://v21.proteinatlas.org/images/18530/41191_A_1_7.jpg

Normal Tissue Figure 8 Lymph node

Non-germinal center cells
Staining:Medium
Intensity: Strong
Quantity: <25%
Location: Cytoplasmic/membranous

* Image credit: Image credit: Human Protein Atlas https://v21.proteinatlas.org/images/18530/41191_A_7_8.jpg

RNA Expression Figure 9 RNA cell line category: Cell line enriched (A-431)

Cell lines ordered by descending RNA expression order.

* Image credit: Image credit: Human Protein Atlas https://v21.proteinatlas.org/ENSG00000146648-EGFR

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Specifications

Immunogen
EGFR expressed on the surface of cells (A431 cells)

Host Species
Human

Derivation
Human

Type
IgG2 - kappa

Specificity
Tested positive against native human antigen

Species Reactivity
Human

Applications
ELISA, IP, FC, FuncS, Neut, IF, ICC

Trade name
vectibix

CAS
339177-26-3

Generic Name
Panitumumab

Biological Half-Life
9.4 days (range: 4-11 days)

ATC Code
L01XC08

DrugBank
DB01269

UNII
6A901E312A

ChEMBL
CHEMBL1201827

MW
144,324.12 g/mol

Related Disease
Colorectal cancers

Product Property

Purity
>95.0%. Determined by analysis by RP-HPLC & analysis by SDS-PAGE.

Storage
4°C. For long term storage, aliquot and store at -20°C. Repeated thawing and freezing must be avoided.

Applications

Application Notes
The EGFR antibody has been reported in applications of ELISA, FC, WB, IF, Inhib.
IF: UM-SCC-22B cell staining study was done as previously described with some modifications (17). Briefly, 70% to 85% confluent tumor cells grown in 35-mm MatTek glass-bottomed culture dishes were fixed using 4% paraformaldehyde for 10 minutes. After blocking with 10% FBS in PBS for 30 minutes, the cells were incubated with panitumumab (5 μg/mL) for 1 hour. The cells were then washed with PBS and stained with FITC-coupled mouse anti–human IgG (hIgG) for 30 minutes. After the final wash with PBS, the cells were visualized using a Leica TCS-NT confocal microscope (Wetzler).

Target

Alternative Names
Panitumumab;vectibix;339177-26-3;ABX-EGF;ABX-EGF;Vectibix;DB01269EGFR;epidermal growth factor receptor;epidermal growth factor receptor (avian erythroblastic leukemia viral (v erb b) oncogene homolog) , ERBB;ERBB1;erythroblastic leukemia viral (v erb b) o

Gene ID
1956

UniProt ID
P00533

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Related Diseases

Bladder Cancer

Non-small Cell Lung Cancer

Pancreatic Cancer

Hepatocellular Carcinoma

Cushing Syndrome

Related Signaling Pathways

JAK-STAT 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

Datasheet - TAB-020.pdf

MSDS

MSDS - TAB-020.pdf

COA

Certificate of Analysis Lookup

To download a Certificate of Analysis, please enter a lot number in the search box below. Note: Certificate of Analysis not available for kit components.

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Protocol & Troubleshooting

Immunofluorescence (IF)

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.

See other products for "Panitumumab"

Afuco™ Anti-EGFR ADCC Recombinant Antibody, ADCC Enhanced (AFC-TAB-020)
This product is an ADCC enhanced antibody produced by our Afuco™ platform. Recombinant monoclonal antibody to Human EGFR. It is a fully human monoclonal antibody specific to the epidermal growth factor receptor (also known as EGF receptor, EGFR, ErbB-1 and HER1 in humans).

DrugMonitor™ Anti-Panitumumab Antibody (VS-1224-YC884)
Panitumumab is a recombinant humanized monoclonal antibody used in the treatment of EGFR-expressing metastatic colorectal carcinoma that is refractory to chemotherapy regimens containing fluoropyrimidine, oxaliplatin, and irinotecan. The DrugMonitor™ Anti-Panitumumab Antibody (VS-1224-YC884) is an anti-drug antibody (ADA) against Panitumumab. This drug-based antibody is raised in mice immunized with the Panitumumab. The anti-Panitumumab antibody may be used in ELISA, pharmacokinetics (PK), and pharmacodynamics (PD) analyses, or serves as a reference standard in ADA assays. It also is an excellent tool for therapeutic drug monitoring, allowing to evaluate the drug efficacy and determine the drug concentration of the Panitumumab in samples.

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