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Mouse Anti-EBNA-1 Recombinant Antibody (clone 3D4)

CAT#: FAMAB-0523WJ

This product is a mouse monoclonal antibody that is specific for EBV EBNA1. This antibody can be used in a variety of applications, such as ELISA, WB, IF.

Published Data Tested Data

Figure 1 Anti-EBNA-1 ELISA.

ELISA

Figure 1 Anti-EBNA-1 ELISA.

3D4 was tested by ELISA, at increasing concentrations, for binding to EBNA-1, BSA, and the cystovirus, polymerase protein, P2 (negative control) coated on Costar plates. Plates were read at 405 nm at 20 minutes. 3D4 shows specificity for EBNA-1 under these conditions. Results are the averages of triplicates and standard deviations are indicated.

Yadav, P., Tran, H., Ebegbe, R., Gottlieb, P., Wei, H., Lewis, R. H., ... & Spatz, L. (2011). Antibodies elicited in response to EBNA-1 may cross-react with dsDNA. PLoS One, 6(1), e14488.

Figure 2 3D4 was tested by ELISA for binding to dsDNA coated on Immulon-2 plates.

ELISA

Figure 2 3D4 was tested by ELISA for binding to dsDNA coated on Immulon-2 plates.

Plates were read at 405 nm at 1 hour. Results in A and B are the average of triplicates and standard deviations are indicated.

Yadav, P., Tran, H., Ebegbe, R., Gottlieb, P., Wei, H., Lewis, R. H., ... & Spatz, L. (2011). Antibodies elicited in response to EBNA-1 may cross-react with dsDNA. PLoS One, 6(1), e14488.

Figure 3 3D4 antibody is of the IgG1 isotype.

ELISA

Figure 3 3D4 antibody is of the IgG1 isotype.

ELISA plates coated with unlabeled anti-IgG1, anti-IgG2a, anti-IgG2b, or anti-IgG3 were incubated with 1.5 µg/ml of 3D4 MAb followed by the respective polyclonal isotype specific antibodies conjugated to alkaline phosphatase.

Yadav, P., Tran, H., Ebegbe, R., Gottlieb, P., Wei, H., Lewis, R. H., ... & Spatz, L. (2011). Antibodies elicited in response to EBNA-1 may cross-react with dsDNA. PLoS One, 6(1), e14488.

Figure 4 Purified 3D4 at a concentration of 10 µg/ml was examined for binding to Crithidia luciliae slides.

IF

Figure 4 Purified 3D4 at a concentration of 10 µg/ml was examined for binding to Crithidia luciliae slides.

Left panel shows the binding of 3D4 to kinetoplasts of Crithidia luciliae (arrow). Right panel; IgG1 isotype control antibody, does not bind specifically to kinetoplasts.

Yadav, P., Tran, H., Ebegbe, R., Gottlieb, P., Wei, H., Lewis, R. H., ... & Spatz, L. (2011). Antibodies elicited in response to EBNA-1 may cross-react with dsDNA. PLoS One, 6(1), e14488.

Figure 5 3D4 adsorbed on dsDNA cellulose beads was completely depleted of antibody with reactivity for dsDNA and EBNA-1 as detected by ELISA.

ELISA

Figure 5 3D4 adsorbed on dsDNA cellulose beads was completely depleted of antibody with reactivity for dsDNA and EBNA-1 as detected by ELISA.

Results represent OD 405 nm values after subtraction of non specific binding to cellulose only beads. Standard deviations of triplicate wells are indicated. Anti-dsDNA and anti-EBNA-1 ELISAs were performed on Immulon-2 and Costar plates respectively and ELISAs were developed when ODs on each plate reached maximal values.

Yadav, P., Tran, H., Ebegbe, R., Gottlieb, P., Wei, H., Lewis, R. H., ... & Spatz, L. (2011). Antibodies elicited in response to EBNA-1 may cross-react with dsDNA. PLoS One, 6(1), e14488.

Figure 6 Post dsDNA cellulose adsorbed 3D4 shows reduced binding to EBNA-1 by Western blot.

Figure 6 Post dsDNA cellulose adsorbed 3D4 shows reduced binding to EBNA-1 by Western blot.

Left panel: Coomassie stained polyacrylamide gel. Right panel: Western blot: filters were immunostained with pre (lanes 2 and 3) or post dsDNA cellulose adsorbed 3D4 MAb (lane 4).

Yadav, P., Tran, H., Ebegbe, R., Gottlieb, P., Wei, H., Lewis, R. H., ... & Spatz, L. (2011). Antibodies elicited in response to EBNA-1 may cross-react with dsDNA. PLoS One, 6(1), e14488.

Figure 7 Binding of 3D4 to EBNA-1 is compared to that of 0211 by ELISA at two different concentrations of MAbs.

ELISA

Figure 7 Binding of 3D4 to EBNA-1 is compared to that of 0211 by ELISA at two different concentrations of MAbs.

3D4 binds more strongly to EBNA-1 than 0211.

Yadav, P., Tran, H., Ebegbe, R., Gottlieb, P., Wei, H., Lewis, R. H., ... & Spatz, L. (2011). Antibodies elicited in response to EBNA-1 may cross-react with dsDNA. PLoS One, 6(1), e14488.

Figure 8 Binding of 3D4 to several antigens is examined by ELISA.

ELISA

Figure 8 Binding of 3D4 to several antigens is examined by ELISA.

3D4 does not show significant binding to other antigens tested.

Yadav, P., Tran, H., Ebegbe, R., Gottlieb, P., Wei, H., Lewis, R. H., ... & Spatz, L. (2011). Antibodies elicited in response to EBNA-1 may cross-react with dsDNA. PLoS One, 6(1), e14488.

Figure 9 Binding of 0211 to several antigens is examined by ELISA.

ELISA

Figure 9 Binding of 0211 to several antigens is examined by ELISA.

0211 binds moderately to Sm but not to the other antigens tested.

Yadav, P., Tran, H., Ebegbe, R., Gottlieb, P., Wei, H., Lewis, R. H., ... & Spatz, L. (2011). Antibodies elicited in response to EBNA-1 may cross-react with dsDNA. PLoS One, 6(1), e14488.

Figure 10 3D4 binds to the carboxyl region while 0211 binds to both the carboxyl and amino regions of EBNA-1.

ELISA

Figure 10 3D4 binds to the carboxyl region while 0211 binds to both the carboxyl and amino regions of EBNA-1.

(B) MAb 3D4 is strongly reactive with LS9 but not LS8 or LS7 as detected by ELISA. (C) MAb 0211 is reactive with LS7, LS8 and LS9 as detected by ELISA. Results in B and C are the average of triplicate wells.

Yadav, P., Tran, H., Ebegbe, R., Gottlieb, P., Wei, H., Lewis, R. H., ... & Spatz, L. (2011). Antibodies elicited in response to EBNA-1 may cross-react with dsDNA. PLoS One, 6(1), e14488.

Figure 11 3D4 recognizes a 148 aa core domain in the carboxyl region of EBNA-1.

ELISA

Figure 11 3D4 recognizes a 148 aa core domain in the carboxyl region of EBNA-1.

3D4 was tested by ELISA for binding to the 3 truncated fragments of EBNA-1. 3D4 binds strongly to all 3 fragments, the smallest being EBNA459-607.

Yadav, P., Tran, H., Ebegbe, R., Gottlieb, P., Wei, H., Lewis, R. H., ... & Spatz, L. (2011). Antibodies elicited in response to EBNA-1 may cross-react with dsDNA. PLoS One, 6(1), e14488.

Figure 12 IgG antibody to EBNA‐1 cross‐reacts with dsDNA.

ELISA

Figure 12 IgG antibody to EBNA‐1 cross‐reacts with dsDNA.

IgG MAb, 3D4 binds strongly to EBNA-1 (A) and cross-reacts with dsDNA (B) as determined by ELISA. ELISAs are representative of three experiments. ELISA values were obtained in triplicate. Error bars indicate standard deviations of triplicates.

Yadav, P., Carr, M. T., Yu, R., Mumbey‐Wafula, A., & Spatz, L. A. (2016). Mapping an epitope in EBNA‐1 that is recognized by monoclonal antibodies to EBNA‐1 that cross‐react with dsDNA. Immunity, inflammation and disease, 4(3), 362-375.

Figure 13 Comparison of the binding of 3D4 and 16D2 to EBNA‐1.

ELISA

Figure 13 Comparison of the binding of 3D4 and 16D2 to EBNA‐1.

The binding of 16D2 and 3D4 to EBNA-1were compared by ELISA. Goat anti-mouse kappa-AP was used as secondary antibody to detect the binding of 16D2 (an IgM MAb) and 3D4 (an IgG MAb) in the same assay. Results represent the average of two experiments. Standard deviation is indicated by error bars.

Yadav, P., Carr, M. T., Yu, R., Mumbey‐Wafula, A., & Spatz, L. A. (2016). Mapping an epitope in EBNA‐1 that is recognized by monoclonal antibodies to EBNA‐1 that cross‐react with dsDNA. Immunity, inflammation and disease, 4(3), 362-375.

Figure 14 Peptide mapping. 3D4 was tested by ELISA for binding to peptides spanning the entire 148 amino acid VBS region.

ELISA

Figure 14 Peptide mapping. 3D4 was tested by ELISA for binding to peptides spanning the entire 148 amino acid VBS region.

Yadav, P., Carr, M. T., Yu, R., Mumbey‐Wafula, A., & Spatz, L. A. (2016). Mapping an epitope in EBNA‐1 that is recognized by monoclonal antibodies to EBNA‐1 that cross‐react with dsDNA. Immunity, inflammation and disease, 4(3), 362-375.

Figure 15 3D4 was examined by ELISA for binding to GC‐15, GL‐15, and PFM‐15.

ELISA

Figure 15 3D4 was examined by ELISA for binding to GC‐15, GL‐15, and PFM‐15.

3D4 was used at a concentration of 5 μg/ml for all above ELISAs. ELISAs are representative of two experiments. ELISA values were obtained in triplicate.

Yadav, P., Carr, M. T., Yu, R., Mumbey‐Wafula, A., & Spatz, L. A. (2016). Mapping an epitope in EBNA‐1 that is recognized by monoclonal antibodies to EBNA‐1 that cross‐react with dsDNA. Immunity, inflammation and disease, 4(3), 362-375.

Figure 16 Peptide inhibition of MAbs binding to EBNA‐1 and dsDNA.

Inhib

Figure 16 Peptide inhibition of MAbs binding to EBNA‐1 and dsDNA.

3D4 (0.05 μg/ml) were incubated with increasing concentrations of peptides PFM-15 or GC-15 and examined by ELISA for binding to EBNA-1 (B). 3D4 (2.5 μg/ml) were incubated with increasing concentrations of peptides PFM-15 or GC-15 and examined by ELISA for binding dsDNA (D).

Yadav, P., Carr, M. T., Yu, R., Mumbey‐Wafula, A., & Spatz, L. A. (2016). Mapping an epitope in EBNA‐1 that is recognized by monoclonal antibodies to EBNA‐1 that cross‐react with dsDNA. Immunity, inflammation and disease, 4(3), 362-375.

Figure 17 MAbs that bind to the carboxyl region of EBNA‐1 but do not cross‐react with dsDNA, recognize a different epitope than the cross‐reactive antibodies.

ELISA

Figure 17 MAbs that bind to the carboxyl region of EBNA‐1 but do not cross‐react with dsDNA, recognize a different epitope than the cross‐reactive antibodies.

(A) The IgG isotype of the non-cross-reactive MAbs 2F8, 14E10, 4F6, and 4B8 were determined by ELISA. (B) The non cross-reactive MAbs, were tested by ELISA at a range of concentrations, for binding to EBNA-1. 3D4 was used as the reference standard. MAbs were tested by ELISA for binding to dsDNA (C), to the amino and carboxyl regions of EBNA-1 (D) and to two truncated carboxyl fragments; aa 452-641 and aa 459-607 (E). MAb concentrations used were 12.5 μg/ml in (C) and 1.25 μg/ml in (D) and (E). ELISAs are representative of two experiments. ELISA values were obtained in triplicate. Error bars represent the standard deviations of triplicates.

Yadav, P., Carr, M. T., Yu, R., Mumbey‐Wafula, A., & Spatz, L. A. (2016). Mapping an epitope in EBNA‐1 that is recognized by monoclonal antibodies to EBNA‐1 that cross‐react with dsDNA. Immunity, inflammation and disease, 4(3), 362-375.

❮

Figure 1 Anti-EBNA-1 ELISA.

Figure 2 3D4 was tested by ELISA for binding to dsDNA coated on Immulon-2 plates.

Figure 3 3D4 antibody is of the IgG1 isotype.

Figure 4 Purified 3D4 at a concentration of 10 µg/ml was examined for binding to Crithidia luciliae slides.

Figure 5 3D4 adsorbed on dsDNA cellulose beads was completely depleted of antibody with reactivity for dsDNA and EBNA-1 as detected by ELISA.

Figure 6 Post dsDNA cellulose adsorbed 3D4 shows reduced binding to EBNA-1 by Western blot.

Figure 7 Binding of 3D4 to EBNA-1 is compared to that of 0211 by ELISA at two different concentrations of MAbs.

Figure 8 Binding of 3D4 to several antigens is examined by ELISA.

Figure 9 Binding of 0211 to several antigens is examined by ELISA.

Figure 10 3D4 binds to the carboxyl region while 0211 binds to both the carboxyl and amino regions of EBNA-1.

Figure 11 3D4 recognizes a 148 aa core domain in the carboxyl region of EBNA-1.

Figure 12 IgG antibody to EBNA‐1 cross‐reacts with dsDNA.

Figure 13 Comparison of the binding of 3D4 and 16D2 to EBNA‐1.

Figure 14 Peptide mapping. 3D4 was tested by ELISA for binding to peptides spanning the entire 148 amino acid VBS region.

Figure 15 3D4 was examined by ELISA for binding to GC‐15, GL‐15, and PFM‐15.

Figure 16 Peptide inhibition of MAbs binding to EBNA‐1 and dsDNA.

Figure 17 MAbs that bind to the carboxyl region of EBNA‐1 but do not cross‐react with dsDNA, recognize a different epitope than the cross‐reactive antibodies.

❯

Figure 1 Mouse Anti-EBNA-1 Recombinant Antibody (clone 3D4) (FAMAB-0523WJ) in SDS-PAGE

SDS-PAGE

Figure 1 Mouse Anti-EBNA-1 Recombinant Antibody (clone 3D4) (FAMAB-0523WJ) in SDS-PAGE

SDS-PAGE analysis of FAMAB-0523WJ in non-reduced (Lane 1) and reduced (Lane 2) conditions. Gel stained for 30 minutes with Coomassie Blue. As a result of different β-mercaptoethanol-reduced proteins (Heavy chain and Light chain) migrate as about 50 kDa and 25 kDa, respectively.

Figure 2 Mouse Anti-EBNA-1 Recombinant Antibody (clone 3D4) (FAMAB-0523WJ) in SEC-HPLC

SEC-HPLC

Figure 2 Mouse Anti-EBNA-1 Recombinant Antibody (clone 3D4) (FAMAB-0523WJ) in SEC-HPLC

SEC-HPLC analysis of FAMAB-0523WJ shows that the purity is more than 95%

Figure 3 Mouse Anti-EBNA-1 Recombinant Antibody (clone 3D4) (FAMAB-0523WJ) in WB

ELISA

Figure 3 Mouse Anti-EBNA-1 Recombinant Antibody (clone 3D4) (FAMAB-0523WJ) in WB

Western blot analysis of FAMAB-0523WJ was performed with Recombinant Epstein-Barr virus Epstein-Barr nuclear antigen 1 (EBNA1), partial-His-Tag.
Lane 1: Reducing antigen (0.1 μg)
Lane 2: Reducing antigen (0.3 μg)
Lane 3: Reducing antigen (0.6 μg)
The secondary antibody: HRP Goat Anti-Mouse IgG (H+L)

Figure 4 Mouse Anti-EBNA-1 Recombinant Antibody (clone 3D4) (FAMAB-0523WJ) in Dot Blot

WB

Figure 4 Mouse Anti-EBNA-1 Recombinant Antibody (clone 3D4) (FAMAB-0523WJ) in Dot Blot

Dot Blot analysis of FAMAB-0523WJ was performed with Recombinant Epstein-Barr virus Epstein-Barr nuclear antigen 1 (EBNA1), partial-His-Tag.
The secondary antibody: HRP Goat Anti-Mouse IgG (H+L)

Figure 5 Mouse Anti-EBNA-1 Recombinant Antibody (clone 3D4) (FAMAB-0523WJ) in ELISA

DB

Figure 5 Mouse Anti-EBNA-1 Recombinant Antibody (clone 3D4) (FAMAB-0523WJ) in ELISA

Antigen: Recombinant Epstein-Barr virus Epstein-Barr nuclear antigen 1 (EBNA1), partial-His-Tag
The secondary antibody: HRP Goat Anti-Mouse IgG (H+L)

❮

Figure 1 Mouse Anti-EBNA-1 Recombinant Antibody (clone 3D4) (FAMAB-0523WJ) in SDS-PAGE

Figure 2 Mouse Anti-EBNA-1 Recombinant Antibody (clone 3D4) (FAMAB-0523WJ) in SEC-HPLC

Figure 3 Mouse Anti-EBNA-1 Recombinant Antibody (clone 3D4) (FAMAB-0523WJ) in WB

Figure 4 Mouse Anti-EBNA-1 Recombinant Antibody (clone 3D4) (FAMAB-0523WJ) in Dot Blot

Figure 5 Mouse Anti-EBNA-1 Recombinant Antibody (clone 3D4) (FAMAB-0523WJ) in ELISA

❯

Specifications

Immunogen
rEBNA-1 in complete Freund's adjuvant

Host Species
Mouse

Derivation
Hybridoma

Type
Mouse IgG, κ

Specificity
EBV EBNA1

Species Reactivity
EBV

Clone
3D4

Applications
ELISA, WB, IF

Related Disease
the autoimmune disease, Systemic Lupus Erythematosus (SLE)

Product Property

Purity
>95% as determined by SDS-PAGE

Concentration
Please refer to the vial label for the specific concentration.

Buffer
PBS

Preservative
No preservatives

Storage
Centrifuge briefly prior to opening vial. Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C long term. Avoid repeated freeze/thaw cycles.

Target

Alternative Names
Epstein-Barr virus EBNA-1; EBNA-1

Gene ID
3783709

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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 - FAMAB-0523WJ.pdf

MSDS

MSDS - FAMAB-0523WJ.pdf

COA

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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 "EBNA-1"

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CAT	Product Name	Application	Type
MHC-YF066	DRB1*01:01/EBV EBNA1 (TSLYNLRRGTALA) MHC Monomer	MHC Multimer
MHC-YF068	DRB1*04:01/EBV EBNA1 (AEGLRALLARSHVER) MHC Monomer	MHC Multimer
MHC-YF072	DRB1*07:01/EBV EBNA1 (VYGGSKTSLYNLRRGTALAI) MHC Monomer	MHC Multimer
MHC-YF073	DRB1*07:01/EBV EBNA1 (PGPLRESIVCYFMVFLQTHI) MHC Monomer	MHC Multimer
MHC-YF403	B*35:01/EBV EBNA-1 (4A) (HPVAEADYFEY) MHC Pentamer	FCM

CAT	Product Name	Application	Type
FAMAB-0524WJ	Mouse Anti-EBNA-1 Recombinant Antibody (clone 16D2)	ELISA, WB, IF	Mouse IgM, κ

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