- 44-790G - Invitrogen Antibodies EGFR antibody

Submitted references 21-Benzylidene digoxin decreases proliferation by inhibiting the EGFR/ERK signaling pathway and induces apoptosis in HeLa cells.

Pessôa MTC, Valadares JMM, Rocha SC, Silva SC, McDermott JP, Sánchez G, Varotti FP, Scavone C, Ribeiro RIMA, Villar JAFP, Blanco G, Barbosa LA
Steroids 2020 Mar;155:108551

HCaRG/COMMD5 inhibits ErbB receptor-driven renal cell carcinoma.

Matsuda H, Campion CG, Fujiwara K, Ikeda J, Cossette S, Verissimo T, Ogasawara M, Gaboury L, Saito K, Yamaguchi K, Takahashi S, Endo M, Fukuda N, Soma M, Hamet P, Tremblay J
Oncotarget 2017 Sep 19;8(41):69559-69576

Tumor endothelial cells promote metastasis and cancer stem cell-like phenotype through elevated Epiregulin in esophageal cancer.

Sun L, Pan J, Yu L, Liu H, Shu X, Sun L, Lou J, Yang Z, Ran Y
American journal of cancer research 2016;6(10):2277-2288

Hydrocortisone and indomethacin negatively modulate EGF-R signaling in human fetal intestine.

Kajanne R, Leppä S, Luukkainen P, Ustinov J, Thiel A, Ristimäki A, Miettinen PJ
Pediatric research 2007 Nov;62(5):570-5

Modulation of survival signaling pathways and persistence of the genotoxic stress as a basis for the synergistic interaction between the atypical retinoid ST1926 and the epidermal growth factor receptor inhibitor ZD1839.

Zanchi C, Zuco V, Lanzi C, Supino R, Zunino F
Cancer research 2005 Mar 15;65(6):2364-72

Aplidin induces apoptosis in human cancer cells via glutathione depletion and sustained activation of the epidermal growth factor receptor, Src, JNK, and p38 MAPK.

Cuadrado A, Garcia-Fernandez LF, Gonzalez L, Suarez Y, Losada A, Alcaide V, Martinez T, Fernandez-Sousa JM, Sanchez-Puelles JM, Munoz A
The Journal of biological chemistry 2003 Jan 3;278(1):241-50

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Peptide Competition. Extracts of A431 cells unstimulated (1) or stimulated with 200 ng/mL EGF for 15 minutes (2-5) were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 5% BSA-TBST buffer overnight at 4°C, then incubated with the EGFR [pY1086] antibody for two hours at room temperature in a 1% BSA-TBST buffer, following prior incubation with: no peptide (1), the non-phosphopeptide corresponding to the phosphopeptide immunogen (2), a generic phosphotyrosine-containing peptide (3), or the phosphopeptide immunogen (4). After washing, the membrane was incubated with goat F(ab’)2 anti-rabbit IgG HRP conjugate (Product # ALI4404) and signals were detected using the Pierce SuperSignal™ method. The data show that only the phosphopeptide corresponding to EGFR [pY1086] blocks the antibody signal, demonstrating the specificity of the antibody. The data also show the induction of EGFR [pY1086] phosphorylation by the addition of EGF to this cell system.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of EGFR (pT1086) was performed by loading 30 µg of A549 (lane1), A549 treated for 10 minutes with 200 ng/mL of EGF (lane2), A431 (lane4) and A431 treated for 10 minutes with 200 ng/mL of EGF (lane5) cell lysate using Novex®NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800), and proteins were transferred to a PVDF membrane and blocked with 5% skim milk for 1 hour at room temperature. EGFR (pT1086) was detected at ~180 kDa using EGFR (pT1086) Rabbit Polyclonal Antibody (Product # 44-790G) at 1:1000 dilution in 5% skim milk at 4°C overnight on a rocking platform. Goat Anti-Rabbit IgG - HRP Secondary Antibody (G21234) at 1:5000 dilution was used and chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of A-431 (Lane 1), A-431 treated with EGF (200 ng/mL for 10 minutes) (Lane 2), A-431 treated with Gefitinib followed by EGF (1uM for 16 hours, 200 ng/mL for 10 minutes) (Lane 3), A-431 treated with Afatinib followed by EGF (0.5 uM for 6 hours, 200 ng/mL for 10 minutes) (Lane 4), A549 (Lane 5), A549 treated with EGF (200 ng/mL for 10 minutes) (Lane 6) and A549 treated with Afatinib followed by EGF (0.5 uM for 6 hours, 200 ng/mL for 10 minutes) (Lane 7). The blot was probed with Anti-Phospho-EGFR (Tyr1086) Rabbit Polyclonal Antibody (Product # 44-790G, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 170 kDa band corresponding to Phospho-EGFR (Tyr1086) was detected and observed to increase upon EGF treatment across cell lines tested. Pre-treatment with EGFR-antagonists, Gefitinib and Afatinib, resulted in inhibition of Phospho-EGFR in A-431 and A549 cell lines. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane using the wet transfer s

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of EGFR was performed using 90% confluent log phase A-431 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Phospho-EGFR(Tyr 1086) Polyclonal Antibody (Product # 44-790G) at the dilution of 1:100 in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing membranous localization. Panel e represents cells treated with antagonist, Afatinib (1uM for 6hrs) followed by EGF (200 ng/mL for 10 minutes), showing no signal. Panel f shows untreated cells with no signal. Panel g represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 HCaRG inhibits tumor growth in a mouse homograft RCC model ( A ) Renca clones were implanted by subcutaneous injection (1 x 10 6 cells/mouse). HCaRG overexpression significantly inhibited tumor growth of homografted Renca cells as seen after 8 days. ++ P < 0.05, * P < 0.01. ( B ) Tumor lysates obtained 28 days after implantation were analyzed by western blot using appropriate antibodies. HCaRG overexpression led to more differentiated tumor cells in experimental RCCs, as indicated by more E-cadherin and less of alphaSMA compared to Neo-controls. Representative blots showed further that HCaRG suppresses EGFR and ErbB3. ErbB2 protein level was not changed by HCaRG overexpression, while its phosphorylation was diminished. The subsequent MAPK and PI3K/AKT pathways were inactivated in HCaRG-RCCs. ( C ) Representative images of H&E stain and immunostaining with two different proliferation markers, PCNA and Ki-67. HCaRG-RCCs showed less cell proliferation with increased multinucleated giant cells relative to Neo-RCCs. The black arrow indicates a multinucleated giant cell. Scale bars, 100 um. ( D ) The mRNA expressions of ErbB receptors and their downstream genes were demonstrated by Real-Time PCR. ++ P < 0.05, + P < 0.005. NS, not significant. ( E ) Representative images and quantitative data of immunostaining with anti-CD34-antibody. HCaRG overexpression markedly decreased CD34-positive microvessels and endothelial cells in experimental RCCs at day 28. * P

44-790G

Antibody data