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biotinylated hepha2 (Thermo Fisher)

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Thermo Fisher biotinylated hepha2
$(A) FACS profiles of the original G4 library (left) and 4.5-fraction yeasts (right) obtained after final screening. Yeast cells were stained with 1 nM <t>hEphA2</t> and a chicken anti-cMyc antibody. (B) Amino acid sequence alignment of monobodies and wild-type Fn3 domain. Sequences of the BC, DE, and FG loops are indicated as black boxes. (C) FACS profiles of yeasts transformed with surface display vectors, pCT-Fn3-EphA2 (E1) and pCT-Fn3-EphA2 (E10). Yeast cells were stained with 10nM hEphA2 and chicken anti-cMyc antibody.$
Biotinylated Hepha2, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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biotinylated hepha2 - by Bioz Stars, 2026-11

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1) Product Images from "Isolation and Characterization of a Monobody with a Fibronectin Domain III Scaffold That Specifically Binds EphA2"

Article Title: Isolation and Characterization of a Monobody with a Fibronectin Domain III Scaffold That Specifically Binds EphA2

Journal: PLoS ONE

doi: 10.1371/journal.pone.0132976

$(A) FACS profiles of the original G4 library (left) and 4.5-fraction yeasts (right) obtained after final screening. Yeast cells were stained with 1 nM hEphA2 and a chicken anti-cMyc antibody. (B) Amino acid sequence alignment of monobodies and wild-type Fn3 domain. Sequences of the BC, DE, and FG loops are indicated as black boxes. (C) FACS profiles of yeasts transformed with surface display vectors, pCT-Fn3-EphA2 (E1) and pCT-Fn3-EphA2 (E10). Yeast cells were stained with 10nM hEphA2 and chicken anti-cMyc antibody.$
Figure Legend Snippet: (A) FACS profiles of the original G4 library (left) and 4.5-fraction yeasts (right) obtained after final screening. Yeast cells were stained with 1 nM hEphA2 and a chicken anti-cMyc antibody. (B) Amino acid sequence alignment of monobodies and wild-type Fn3 domain. Sequences of the BC, DE, and FG loops are indicated as black boxes. (C) FACS profiles of yeasts transformed with surface display vectors, pCT-Fn3-EphA2 (E1) and pCT-Fn3-EphA2 (E10). Yeast cells were stained with 10nM hEphA2 and chicken anti-cMyc antibody.

Techniques Used: Staining, Sequencing, Transformation Assay

The indicated concentrations of hEphA2 protein were incubated with yeasts expressing each monobody. After a simple wash, yeast cells were stained with mouse anti-hEphA2 and chicken anti-cMyc antibodies, followed by staining with Alexa 488-conjugated anti-mouse and Alexa 555-conjugated anti-chicken secondary antibodies. The mean fluorescence values of each sample were measured by FACS. Affinities (Kd values) were obtained by determining the mean ratio of Alexa 488/Alexa 555 fluorescence versus hEphA2 concentration. The results are representatives of at least three independent experiments.

Figure Legend Snippet: The indicated concentrations of hEphA2 protein were incubated with yeasts expressing each monobody. After a simple wash, yeast cells were stained with mouse anti-hEphA2 and chicken anti-cMyc antibodies, followed by staining with Alexa 488-conjugated anti-mouse and Alexa 555-conjugated anti-chicken secondary antibodies. The mean fluorescence values of each sample were measured by FACS. Affinities (Kd values) were obtained by determining the mean ratio of Alexa 488/Alexa 555 fluorescence versus hEphA2 concentration. The results are representatives of at least three independent experiments.

Techniques Used: Incubation, Expressing, Staining, Fluorescence, Concentration Assay

Streptavidin-coated magnetic beads bound with biotinylated human IgG, lysozyme, or hEphA2 were incubated with 10 nM monobody proteins with His6 (h) or AMH tags at room temperature. The bound monobodies were stained with a FITC-conjugated anti-His-tag antibody and analyzed by FACS. The results are representatives of at least three independent experiments.

Figure Legend Snippet: Streptavidin-coated magnetic beads bound with biotinylated human IgG, lysozyme, or hEphA2 were incubated with 10 nM monobody proteins with His6 (h) or AMH tags at room temperature. The bound monobodies were stained with a FITC-conjugated anti-His-tag antibody and analyzed by FACS. The results are representatives of at least three independent experiments.

Techniques Used: Magnetic Beads, Incubation, Staining

The indicated concentrations of AMH-tagged monobody proteins were incubated for 2 hr on 96-well plates coated with recombinant Eph receptor proteins. The amounts of bound monobody proteins were measured in an ELISA reader after staining with horseradish peroxidase-conjugated anti-cMyc antibodies. Data represent mean ± S.D., and asterisks (*) indicate a significant difference compared hEphA2 and other EphA series (*, P < 0.05 or **, P < 0.01). The results are representatives of at least three independent experiments.

Figure Legend Snippet: The indicated concentrations of AMH-tagged monobody proteins were incubated for 2 hr on 96-well plates coated with recombinant Eph receptor proteins. The amounts of bound monobody proteins were measured in an ELISA reader after staining with horseradish peroxidase-conjugated anti-cMyc antibodies. Data represent mean ± S.D., and asterisks (*) indicate a significant difference compared hEphA2 and other EphA series (*, P < 0.05 or **, P < 0.01). The results are representatives of at least three independent experiments.

Techniques Used: Incubation, Recombinant, Enzyme-linked Immunosorbent Assay, Staining

(A) Western blotting for hEphA2. Cell lysates were separated by 10% SDS-PAGE and transferred onto nitrocellulose membranes. The proteins were detected with an anti-hEphA2 antibody. (B) Surface expression of hEphA2. Cells (2 × 10 5 ) were stained with a mouse anti-hEphA2 antibody and FITC-conjugated rat anti-mouse IgG antibody. Black line, unstained cells; red line, only second Ab; blue line, anti-EphA2. (C) Quantitation of hEphA2 on the cell surface. Concentrations of hEphA2 on the cell surface were measured against an anti-rat Bangs bead standard. Data represent mean ± S.D., and asterisks (*) indicate a significant difference compared SKBR3 and PC3 cells (*, P < 0.001). (D) Measurement of E1 monobody binding to cells. Cells were incubated with 100 nM Cy5.5-conjugated E1h monobody and analyzed by FACS. Black lines, unstained cells; red lines, E1h-Cy5.5. (F) Fluorescence microscopy images of cells stained with an anti-hEphA2 antibody or E1h-Cy5.5 monobody. Cell nuclei were stained with 4',6-diamidino-2-phenylindole (DAPI). Magnification of the inset with indivisual and merged images is given in the lower low. Scale bar, 20 μm. The results are representatives of at least three independent experiments.

Figure Legend Snippet: (A) Western blotting for hEphA2. Cell lysates were separated by 10% SDS-PAGE and transferred onto nitrocellulose membranes. The proteins were detected with an anti-hEphA2 antibody. (B) Surface expression of hEphA2. Cells (2 × 10 5 ) were stained with a mouse anti-hEphA2 antibody and FITC-conjugated rat anti-mouse IgG antibody. Black line, unstained cells; red line, only second Ab; blue line, anti-EphA2. (C) Quantitation of hEphA2 on the cell surface. Concentrations of hEphA2 on the cell surface were measured against an anti-rat Bangs bead standard. Data represent mean ± S.D., and asterisks (*) indicate a significant difference compared SKBR3 and PC3 cells (*, P < 0.001). (D) Measurement of E1 monobody binding to cells. Cells were incubated with 100 nM Cy5.5-conjugated E1h monobody and analyzed by FACS. Black lines, unstained cells; red lines, E1h-Cy5.5. (F) Fluorescence microscopy images of cells stained with an anti-hEphA2 antibody or E1h-Cy5.5 monobody. Cell nuclei were stained with 4',6-diamidino-2-phenylindole (DAPI). Magnification of the inset with indivisual and merged images is given in the lower low. Scale bar, 20 μm. The results are representatives of at least three independent experiments.

Techniques Used: Western Blot, SDS Page, Expressing, Staining, Quantitation Assay, Binding Assay, Incubation, Fluorescence, Microscopy

Protein Concentration:

Article Title: OT-Curtains: an approach for studying protein interactions with DNA ends using optical tweezers and confocal fluorescence microscopy
Article Snippet: .. In some experiments, bio-AddAB was pre-incubated with streptavidin-conjugated Qdot525 (Thermo Fisher Scientific, Q10143MP) at a 1:2 (AddAB:Qdot) molar ratio for 10 min on ice before dilution with experimental buffer to a final protein concentration of 10 nM (referred to as QD525-AddAB). ..

other:

Article Title: Interacting proteins of AMPK studied using TurboID proximity labeling technology
Article Snippet: Dynabeads MyOne streptavidin C1 (cat. no. 65001), biotin (cat. no. B20656 ) and liposomal transfection reagent (Lipofectamine 2000; cat. no. 11668500) were obtained from Thermo Fisher Scientific, Inc. RIPA buffer (cat. no. G2002-100ML), DAPI (cat. no. G1012-100ML) and anti-fluorescence quenching sealing agent (cat. no. G1401-25ML) were purchased from Wuhan Servicebio Technology Co., Ltd.

Article Title: A conserved mammalian mecciRNA, mecciATP6, regulates mitochondrial homeostasis through interaction with HNRNPA3
Article Snippet: For smFISH and immunofluorescence (IF) analyses, the colocalization ratio between smFISH/IF signals and mitochondria was quantified using the Coloc2 plugin in ImageJ.

Article Title: Targeting Mettl8-Tcf1 axis promotes CD8 + T PEX differentiation and antitumor immunity
Article Snippet: The DNA ends were blunted and labeled with biotin by Klenow DNA polymerase in the presence of dCTP, dGTP, dTTP, and biotin-14-dATP, followed by ligation using T4 DNA ligase (NEB).

Transfection:

Article Title: CircEif3c/miR-96–5p/PHF20L1/MEOX2 axis in perivascular preadipocyte exosomes mediates fibroblast dysfunction and vascular remodeling
Article Snippet: Biotin-labeled miRNAs were transfected into cells using Lipofectamine 3000 (#L3000-001, Invitrogen, Thermo Fisher Scientific, USA), and cell lysates were subsequently collected. .. Cell lysates were collected 48 h after transfection and incubated with streptavidin magnetic beads (#65306, Invitrogen, USA) for 2 h. After centrifugation to wash the beads, the pulled-down miRNAs were extracted using Trizol (#15596018, Invitrogen, USA) and subjected to qRT-PCR. .. Cell lysates were collected 48 h after transfection and incubated with streptavidin magnetic beads (#65306, Invitrogen, USA) for 2 h. After centrifugation to wash the beads, the pulled-down miRNAs were extracted using Trizol (#15596018, Invitrogen, USA) and subjected to qRT-PCR.

Incubation:

Article Title: HSF1 and YTHDC1 enhance DNA repair under heat stress by promoting repair-associated gene expression
Article Snippet: The samples were washed three more times in PBS containing 0.1% Tween 20, incubated with 4′,6-diamidino-2-phenylindole (DAPI) (1 μg/ml, Beyotime, C1002) for 5 min, and sealed. .. In addition, cells were incubated with streptavidin 488 (Invitrogen, S11223 ) simultaneously with secondary antibodies. .. Images were acquired using a Leica STELLARIS 5 microscope or a Leica TCS SP8 STED 3X microscope for high-resolution STED imaging.

Article Title: CircSMAD4 shapes matrix-remodeling TAMs in lung adenocarcinoma
Article Snippet: Samples were acquired on a BD LSRFortessa flow cytometer. .. Biotinylated circSMAD4 sense RNA (with antisense RNA as a negative control) was generated by in vitro transcription using Biotin-16-UTP (Beyotime, Cat# D7336), incubated with macrophage lysates, and captured using streptavidin beads (Invitrogen, Cat# 88816). ..

Magnetic Beads:

Centrifugation:

Cell-Signaling:

Article Title: Divalent siRNA for prion disease
Article Snippet: .. The assay uses antibodies EP1802Y (Abcam, ab52604) for capture and in-house biotinylated 8H4 (Abcam ab61409) for detection, followed by streptavidin–horseradish peroxidase (Thermo Fisher Scientific, 21130) and TMB (Cell Signaling, 7004P4). ..

Negative Control:

Generated:

In Vitro:

Image Search Results

(A) Western blot analysis of hEphA2. Lysates of PC3 and HeLa cells (8 × 10 5 ) were separated by SDS-PAGE and transferred to nylon membranes. Membranes were stained with rabbit anti-hEphA2 or β-actin antibodies. (B) Flow cytometric analysis of cells. Cells (5 × 10 5 ) were stained with mouse anti-hEphA2 and Alexa 488-conjugated anti-mouse IgG antibodies (anti-hEphA2), and fluorescence was measured by flow cytometry. Unstained cells or cells stained with Alexa 488-conjugated anti-mouse IgG antibody (2 nd Ab) alone were used as controls.

Journal: PLoS ONE

Article Title: Engineering of monobody conjugates for human EphA2-specific optical imaging

doi: 10.1371/journal.pone.0180786

Figure Lengend Snippet: (A) Western blot analysis of hEphA2. Lysates of PC3 and HeLa cells (8 × 10 5 ) were separated by SDS-PAGE and transferred to nylon membranes. Membranes were stained with rabbit anti-hEphA2 or β-actin antibodies. (B) Flow cytometric analysis of cells. Cells (5 × 10 5 ) were stained with mouse anti-hEphA2 and Alexa 488-conjugated anti-mouse IgG antibodies (anti-hEphA2), and fluorescence was measured by flow cytometry. Unstained cells or cells stained with Alexa 488-conjugated anti-mouse IgG antibody (2 nd Ab) alone were used as controls.

Article Snippet: Cells (1 × 10 4 ) were cultured on cover glasses on 6-well plates for 24 h. To detect bound E1-Rluc8, cells were incubated with 25.5 nM E1-Rluc8 and rabbit anti-hEphA2 antibody (1:10000 dilution, Santa Cruz Biotechnology), in 3 mL of culture medium containing 3% FBS at 37°C for 2 h. After the plates were washed with 1% BSA in PBS-T, they were incubated in FITC-conjugated anti-6×His (1:1000 dilution) and Alexa 555-conjugated anti-rabbit IgG antibodies (1:1000 dilution) for 2 h in the dark at room temperature (RT).

Techniques: Western Blot, SDS Page, Staining, Fluorescence, Flow Cytometry

(A) Flow cytometric analysis of cells treated with E1-Rluc8 and E1-EGFP. PC3 and HeLa cells (5 × 10 5 ) detached from culture plates were treated with E1-Rluc8 and E1-EGFP. The cells treated with E1-Rluc8 were stained with FITC-conjugated mouse anti-6×His antibody. Unstained cells or cells stained with FITC-conjugated secondary antibody (2 nd Ab) alone were used as controls. (B) Fluorescence microscopy images of the cells. PC3 and HeLa cells were stained with mouse anti-hEphA2 antibody and E1-Rluc8, and then detected with Alexa 555-conjugated anti-mouse IgG (red) and FITC-conjugated anti-6×His antibodies (green). Cell nuclei were stained with DAPI (blue). Scale bar, 10 μm. (C) Fluorescence microscopy images of xenograft tumor tissues. PC3 tumor tissues from transplanted nude mice were stained with the antibody combinations used in (B). Scale bar, 10 μm.

Journal: PLoS ONE

Article Title: Engineering of monobody conjugates for human EphA2-specific optical imaging

doi: 10.1371/journal.pone.0180786

Figure Lengend Snippet: (A) Flow cytometric analysis of cells treated with E1-Rluc8 and E1-EGFP. PC3 and HeLa cells (5 × 10 5 ) detached from culture plates were treated with E1-Rluc8 and E1-EGFP. The cells treated with E1-Rluc8 were stained with FITC-conjugated mouse anti-6×His antibody. Unstained cells or cells stained with FITC-conjugated secondary antibody (2 nd Ab) alone were used as controls. (B) Fluorescence microscopy images of the cells. PC3 and HeLa cells were stained with mouse anti-hEphA2 antibody and E1-Rluc8, and then detected with Alexa 555-conjugated anti-mouse IgG (red) and FITC-conjugated anti-6×His antibodies (green). Cell nuclei were stained with DAPI (blue). Scale bar, 10 μm. (C) Fluorescence microscopy images of xenograft tumor tissues. PC3 tumor tissues from transplanted nude mice were stained with the antibody combinations used in (B). Scale bar, 10 μm.

Techniques: Staining, Fluorescence, Microscopy

(A) Measurement of luminescence in mice injected with different concentrations of E1-Rluc8. PC3 and HeLa cells were transplanted into 6-week-old Balb/c nude mice via subcutaneous injection (n = 3). After tumor formation, the indicated amounts of E1-Rluc8 and coelenterazine were intravenously injected via the tail vein, and luminescence images were acquired at the indicated times with the NightOWL in vivo imaging system (left). The luminescence intensities measured in tumor areas at 6 h were graphed (right). (B) Luminescence maintenance in mice injected with E1-Rluc8 for 24 h. E1-Rluc8 (60 μg) and coelenterazine were intravenously injected via the tail vein into PC3 tumor mice (n = 5), and luminescence images were obtained at the indicated times (left). The luminescence intensities in tumor tissues were measured with the NightOWL in vivo imaging system and graphed (right). (C) Detection of the remaining E1-Rluc8 in PC3 tumor tissues. Tumor tissues were collected at the indicated times from mice injected with E1-Rluc8 and stained with FITC-conjugated anti-6×His antibodies (green). hEphA2 was stained with mouse anti-hEphA2 antibody and Alexa 555-conjugated anti-mouse IgG (red). Cell nuclei were stained with DAPI (blue). Scale bar, 10 μm.

Journal: PLoS ONE

Article Title: Engineering of monobody conjugates for human EphA2-specific optical imaging

doi: 10.1371/journal.pone.0180786

Figure Lengend Snippet: (A) Measurement of luminescence in mice injected with different concentrations of E1-Rluc8. PC3 and HeLa cells were transplanted into 6-week-old Balb/c nude mice via subcutaneous injection (n = 3). After tumor formation, the indicated amounts of E1-Rluc8 and coelenterazine were intravenously injected via the tail vein, and luminescence images were acquired at the indicated times with the NightOWL in vivo imaging system (left). The luminescence intensities measured in tumor areas at 6 h were graphed (right). (B) Luminescence maintenance in mice injected with E1-Rluc8 for 24 h. E1-Rluc8 (60 μg) and coelenterazine were intravenously injected via the tail vein into PC3 tumor mice (n = 5), and luminescence images were obtained at the indicated times (left). The luminescence intensities in tumor tissues were measured with the NightOWL in vivo imaging system and graphed (right). (C) Detection of the remaining E1-Rluc8 in PC3 tumor tissues. Tumor tissues were collected at the indicated times from mice injected with E1-Rluc8 and stained with FITC-conjugated anti-6×His antibodies (green). hEphA2 was stained with mouse anti-hEphA2 antibody and Alexa 555-conjugated anti-mouse IgG (red). Cell nuclei were stained with DAPI (blue). Scale bar, 10 μm.

Techniques: Injection, In Vivo Imaging, Staining

Phage supernatants of the seven clones with different loop DE sequences were tested by ELISA on plates coated with mEphA2-Fc, hEphA2, DR6-Fc and BSA to asses specificity of binding and crossreactivity.

Journal: PLoS ONE

Article Title: High Affinity Binders to EphA2 Isolated from Abdurin Scaffold Libraries; Characterization, Binding and Tumor Targeting

doi: 10.1371/journal.pone.0135278

Figure Lengend Snippet: Phage supernatants of the seven clones with different loop DE sequences were tested by ELISA on plates coated with mEphA2-Fc, hEphA2, DR6-Fc and BSA to asses specificity of binding and crossreactivity.

Article Snippet: Chinese Hamster Ovary K1 cells (CHOK1), (ATCC, Manassas, Virginia, USA) were transfected with transfection-ready hEphA2 cDNA (OriGene Technologies, Rockville, Maryland, USA) according to the manufacturer’s instructions.

Techniques: Clone Assay, Enzyme-linked Immunosorbent Assay, Binding Assay

Phage supernatants of the seven clones with different loop DE sequences were tested by cell-based ELISA on HEK293 cells expressing doxycycline-inducible hEphA2. All the Abdurin variants specifically recognize the hEphA2 receptor in its native conformation on cells. Assays were done in the absence or presence of doxycycline with concentrations of the phage ranging from 25 ng/ml to 100 ng/ml.

Journal: PLoS ONE

Article Title: High Affinity Binders to EphA2 Isolated from Abdurin Scaffold Libraries; Characterization, Binding and Tumor Targeting

doi: 10.1371/journal.pone.0135278

Figure Lengend Snippet: Phage supernatants of the seven clones with different loop DE sequences were tested by cell-based ELISA on HEK293 cells expressing doxycycline-inducible hEphA2. All the Abdurin variants specifically recognize the hEphA2 receptor in its native conformation on cells. Assays were done in the absence or presence of doxycycline with concentrations of the phage ranging from 25 ng/ml to 100 ng/ml.

Techniques: Clone Assay, In-Cell ELISA, Expressing

Affinity matured binders to EphA2, selected by CIS display, were tested in end-point titration ELISA assays in 96-well plates coated with hEphA2-Fc (a), mEphA2-Fc (b) or strepavidin (c). The purified matured Abdurin clones D2, G7, B6 and B11 were tested alongside the purified non-matured parental EphA2 binding clones selected by phage display, E10par and H3par, as detected by anti-FLAG M2-HRP conjugated antibody. Binding of Abdurin binders and the truncated wild-type human CH2 (shWTCH2) to CHO cells transfected with human EphA2 (d) or to non-transfected cells (e), was assessed by FACS analysis.

Journal: PLoS ONE

Article Title: High Affinity Binders to EphA2 Isolated from Abdurin Scaffold Libraries; Characterization, Binding and Tumor Targeting

doi: 10.1371/journal.pone.0135278

Figure Lengend Snippet: Affinity matured binders to EphA2, selected by CIS display, were tested in end-point titration ELISA assays in 96-well plates coated with hEphA2-Fc (a), mEphA2-Fc (b) or strepavidin (c). The purified matured Abdurin clones D2, G7, B6 and B11 were tested alongside the purified non-matured parental EphA2 binding clones selected by phage display, E10par and H3par, as detected by anti-FLAG M2-HRP conjugated antibody. Binding of Abdurin binders and the truncated wild-type human CH2 (shWTCH2) to CHO cells transfected with human EphA2 (d) or to non-transfected cells (e), was assessed by FACS analysis.

Techniques: Titration, Enzyme-linked Immunosorbent Assay, Purification, Clone Assay, Binding Assay, Transfection

Journal: PLoS ONE

Article Title: High Affinity Binders to EphA2 Isolated from Abdurin Scaffold Libraries; Characterization, Binding and Tumor Targeting

doi: 10.1371/journal.pone.0135278

Figure Lengend Snippet: K D measured using Biacore 3000.

Techniques:

Affinity measurement of binding to hEphA2 transfected cells.

Journal: PLoS ONE

Article Title: High Affinity Binders to EphA2 Isolated from Abdurin Scaffold Libraries; Characterization, Binding and Tumor Targeting

doi: 10.1371/journal.pone.0135278

Figure Lengend Snippet: Affinity measurement of binding to hEphA2 transfected cells.

Techniques: Binding Assay, Transfection

A) HEK293-hEphA2 cells were seeded in 96 well plates and either, not treated (top panel), or treated (bottom panel) with 100 nM doxycycline for 24 hours at 37°C to induce over-expression of the transfected EphA2 receptor. Cells were incubated with 1μg/well B11 for 30 minutes at 4°C after which time cells were fixed in 2% PFA. B11 binding was monitored by anti-FLAG antibody and anti-mouse AF488 secondary antibody. B) PC-3 cells seeded in 96 well plates were incubated with 1μg/well B11 and 10 μg/well of the anti-EphA2 monoclonal antibody (mAb) (molar ratio 1:1; bottom panel) or an unrelated mAb (top panel) for 30 minutes at 4°C followed by 1 hour incubation at 37°C to allow internalization. Cells were then fixed in 2% PFA and B11 binding monitored by anti-FLAG-AF488 antibody. Nuclei were stained with DAPI. Images were acquired at IN Cell 2000.

Journal: PLoS ONE

Article Title: High Affinity Binders to EphA2 Isolated from Abdurin Scaffold Libraries; Characterization, Binding and Tumor Targeting

doi: 10.1371/journal.pone.0135278

Figure Lengend Snippet: A) HEK293-hEphA2 cells were seeded in 96 well plates and either, not treated (top panel), or treated (bottom panel) with 100 nM doxycycline for 24 hours at 37°C to induce over-expression of the transfected EphA2 receptor. Cells were incubated with 1μg/well B11 for 30 minutes at 4°C after which time cells were fixed in 2% PFA. B11 binding was monitored by anti-FLAG antibody and anti-mouse AF488 secondary antibody. B) PC-3 cells seeded in 96 well plates were incubated with 1μg/well B11 and 10 μg/well of the anti-EphA2 monoclonal antibody (mAb) (molar ratio 1:1; bottom panel) or an unrelated mAb (top panel) for 30 minutes at 4°C followed by 1 hour incubation at 37°C to allow internalization. Cells were then fixed in 2% PFA and B11 binding monitored by anti-FLAG-AF488 antibody. Nuclei were stained with DAPI. Images were acquired at IN Cell 2000.

Techniques: Over Expression, Transfection, Incubation, Binding Assay, Staining

Review

biotinylated hepha2 (Thermo Fisher)

Structured Review

Images

1) Product Images from "Isolation and Characterization of a Monobody with a Fibronectin Domain III Scaffold That Specifically Binds EphA2"

Related Articles

Protein Concentration:

other:

Transfection:

Incubation:

Magnetic Beads:

Centrifugation:

Cell-Signaling:

Negative Control:

Generated:

In Vitro:

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