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75352
Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit #75352

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Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 1

Western blot analysis of extracts from various cell lines using ACE2 Antibody (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 2

Confocal immunofluorescent analysis of Caco-2 (positive, left) and HT-1080 (negative, right) cells using DPP4 (D6D8K) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 3

Flow cytometric analysis of peripheral blood using CD13/APN (D6V1W) Rabbit mAb (blue) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red). Anti-Rabbit IgG (H+L), F(ab')2Fragment (Alexa Fluor® 488 Conjugate) #4412 was used a secondary antibody.

   

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 4

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Basigin/EMMPRIN (E1S1V) Rabbit mAb.

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 5

Confocal immunofluorescent analysis of HeLa cells using EEA1 (C45B10) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 6

Western blot analysis of extracts from K-562 cells, CD4+ T cells, and CD8+ T cells using IFITM1 Antibody.

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 7

Flow cytometric analysis of Daudi cells (blue) and U266 cells (green) using Cathepsin B (D1C7Y) XP® Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 8

Western blot analysis of extracts from various cell lines using IFITM3 (D8E8G) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 9

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using IFITM3 (D8E8G) XP® Rabbit mAb.

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 10

Western blot analysis of extracts from HeLa, NIH/3T3, C6 and COS cells, using VCP (7F3) Rabbit mAb.

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 11

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 12

Western blot analysis of extracts from human testis and kidney tissue using ACE2 Antibody.

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 13

Immunoprecipitation of DPP4 protein from LoVo cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is DPP4 (D6D8K) Rabbit mAb. Western blot analysis was performed using DPP4 (D6D8K) Rabbit mAb.

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 14

Confocal immunofluorescent analysis of PANC-1 (left) or HEK293 (right) cells using CD13/APN (D6V1W) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 15

Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using Basigin/EMMPRIN (E1S1V) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 16

Western blot analysis of extracts from various cell lines using EEA1 (C45B10) Rabbit mAb.

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 17

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing full-length human IFITM1 (hIFITM1; +), using IFITM1 Antibody.

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 18

Confocal immunofluorescent analysis of Malme-3M (left) and MCF7 (right) cells using Cathepsin (D1C7Y) XP® Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 19

Immunohistochemical analysis of paraffin-embedded normal human lymph node using IFITM3 (D8E8G) XP® Rabbit mAb.

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 20

Western blot analysis of extracts from Lovo, Caco-2 and IGROV-1 cell lines using DPP4 (D6D8K) Rabbit mAb.

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 21

Immunohistochemical analysis of paraffin-embedded normal human kidney using CD13/APN (D6V1W) Rabbit mAb.

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 22

Western blot analysis of extracts from various cell lines using Basigin/EMMPRIN (E1S1V) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 23

Western blot analysis of extracts from HeLa cells, untreated (-) or treated with Human Interferon-α1 (hIFN-α1) #8927 (10 ng/ml, 16 hr; +), using IFITM1 Antibody (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 24

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Cathepsin B (D1C7Y) XP(R) Rabbit mAb.

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 25

Immunohistochemical analysis of paraffin-embedded HeLa (left) and RPMI 8226 (right) cell pellets using IFITM3 (D8E8G) XP® Rabbit mAb.

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 26

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using CD13/APN (D6V1W) Rabbit mAb.

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 27

Western blot analysis of extracts from Ramos cells, untreated (-) or PNGase F-treated (+), using Basigin/EMMPRIN (E1S1V) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 28

Immunohistochemical analysis of paraffin-embedded normal human kidney using Cathepsin B (D1C7Y) XP(R) Rabbit mAb.

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 29

Immunohistochemical analysis of paraffin-embedded normal human spleen using IFITM3 (D8E8G) XP® Rabbit mAb.

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 30

Immunohistochemical analysis of paraffin-embedded PANC-1 (left) and HEK-293 (right) cell pellets using CD13/APN (D6V1W) Rabbit mAb.

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 31

Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using Cathepsin B (D1C7Y) XP(R) Rabbit mAb.

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 32

Immunoprecipitation of CD13/APN from PANC-1 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is CD13/APN (D6V1W) Rabbit mAb. Western blot analysis was performed using CD13/APN (D6V1W) Rabbit mAb.

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 33

Western blot analysis of extracts from 293T cells untransfected (-), or transfected with a construct expressing Myc-tagged full-length human IFITM3 (hIFITM3-Myc; +), using IFITM3 (D8E8G) XP® Rabbit mAb.

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 34

Western blot analysis of extracts from OVCAR8, SK-MEL-2, mouse kidney, and rat liver using CD13/APN (D6V1W) Rabbit mAb (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 35

Western blot analysis of extracts from various cell lines using Cathepsin B (D1C7Y) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit: Image 36

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing full-length human Cathepsin B (hCTSB; +) or mouse Cathespin B (mCTSB; +) using Cathepsin B (D1C7Y) XP® Rabbit mAb.

To Purchase # 75352T
製品番号 サイズ 価格 在庫
75352T
1 Kit  (9 x 20 µl)

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
ACE2 Antibody 4355 20 µl
  • WB
  • IP
H 120-135 Rabbit 
DPP4/CD26 (D6D8K) Rabbit mAb 67138 20 µl
  • WB
  • IP
  • IF
H 90, 120 Rabbit IgG
CD13/APN (D6V1W) Rabbit mAb 32720 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H M R 160 Rabbit IgG
Basigin/EMMPRIN (E1S1V) Rabbit mAb 13287 20 µl
  • WB
  • IHC
H 38-58 Rabbit IgG
EEA1 (C45B10) Rabbit mAb 3288 20 µl
  • WB
  • IP
  • IF
H M R 170 Rabbit IgG
IFITM1 Antibody 13126 20 µl
  • WB
H 14 Rabbit 
Cathepsin B (D1C7Y) XP® Rabbit mAb 31718 20 µl
  • WB
  • IHC
  • IF
  • F
H M R 44, 27, 24 Rabbit IgG
IFITM3 (D8E8G) XP® Rabbit mAb 59212 20 µl
  • WB
  • IP
  • IHC
H 15 Rabbit IgG
VCP (7F3) Rabbit mAb 2649 20 µl
  • WB
H M R Mk 89 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

The Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit provides an economical means of detecting key host cell proteins involved in the attachment and cellular entry of coronaviruses. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody in the Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit detects endogenous levels of its target protein. ACE2 Antibody recognizes endogenous levels of glycosylated ACE2 protein. This antibody also cross-reacts with 55 and 75 kDa proteins of unknown origin in some cells. IFITM1 Antibody recognizes endogenous levels of total IFITM1 protein. This antibody does not cross-react with IFITM2 or IFITM3 proteins. Cathepsin B (D1C7Y) XP® Rabbit mAb recognizes endogenous levels of total cathepsin B protein. This antibody detects the heavy chain subunit of cathepsin B. IFITM3 (D8E8G) XP® Rabbit mAb recognizes endogenous levels of total IFITM3 protein. This antibody does not cross-react with IFITM1 or IFITM2 proteins.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu491 of human DPP4 protein, Ala254 of human EMMPRIN protein, Ser70 of human EEA1 protein, Val5 of human IFITM3 protein, and a synthetic peptide corresponding to the sequence of human VCP. Monoclonal antibodies are produced by immunizing animals with a recombinant protein specific to the carboxy terminus of human CD13/APN protein and a recombinant protein specific to the heavy chain subunit of human cathepsin B protein. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human ACE2 protein and surrounding Pro20 of human IFITM1 protein. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

Background

Coronaviruses are a group of viruses that contain single-stranded, positive-sense RNA genomes. Several members of this group, which include severe acute respiratory syndrome coronaviruses (SARS-CoV and SARS-CoV-2) and Middle East respiratory syndrome coronavirus (MERS-CoV), are highly pathogenic and have caused significant disease outbreaks in human hosts. In order for human coronaviruses to transcribe and replicate their genomes within host cells, they must first attach and gain entry into host cells using a variety of cell surface receptors and components of the endocytic machinery.

ACE2 is a carboxypeptidase that catalyses the conversion of angiotensin I to angiotensin 1-9, or of angiotensin II to the vasodilator angiotensin 1-7 (1). Research studies have identified ACE2 as the receptor for SARS and SARS-CoV-2 coronaviruses (2-4).

DPP4 (CD26) is a type II transmembrane glycoprotein expressed ubiquitously in most tissues and different cell types (5,6). In addition to its peptidase activity, DPP4 interacts with multiple important cell surface ligands, such as adenosine deaminase, fibronectin, and IGF2 receptor to influence processes like T cell activation, cell migration, and proliferation (7). Research studies have shown that DPP4 serves as a cellular receptor for the MERS-CoV spike protein (8).

Aminopeptidase N (APN, CD13) is a widely expressed, membrane-bound proteolytic enzyme that breaks down peptides during digestion, cleaves cell surface antigens during antigen presentation, and acts as a receptor for human viruses, including several coronaviruses. This multifunctional protein is implicated in the regulation of many biological processes, including angiogenesis, cell proliferation, cell migration, inflammation, and immune response (9,10).

Basigin (EMMPRIN, CD147) is a type I integral membrane receptor protein belonging to the immunoglobulin superfamily (11). Multiple functions have been ascribed to Basigin; foremost among these is stimulating the secretion of extracellular matrix metalloproteinases by adjacent fibroblasts, a function which has been implicated in promoting tumor progression (12-14). Research studies have suggested that Basigin serves as a novel host cell surface receptor for SARS-CoV-2 (15).

EEA1 is an early endosomal marker and a Rab5 effector protein essential for early endosomal membrane fusion and trafficking (16,17). Research studies have shown that efficient coronavirus host cell entry and replication relies upon early endosomes containing EEA1 (18).

Interferon-induced transmembrane protein (IFITM) family members are composed of short amino- and carboxy-termini, two transmembrane domains, and a cytoplasmic domain (19). The primary function of IFITM family proteins appears to be viral restriction, as IFITM proteins inhibit cytosolic entry of coronaviruses by preventing fusion of viral and host membranes (20,21).

Valosin-containing protein (VCP) is a highly conserved and abundant 97 kDa protein that belongs to the AAA family of proteins. These protein complexes participate in many cellular functions, including vesicle transport and fusion, fragmentation and reassembly of the golgi stacks during mitosis, nuclear envelope formation and spindle disassembly following mitosis, cell cycle regulation, DNA damage repair, apoptosis, B and T cell activation, NF-κB-mediated transcriptional regulation, endoplasmic reticulum (ER)-associated degradation, and protein degradation (22). Research studies have shown that VCP facilitates the release of some coronaviruses from the early endosomal compartment (23).

Cathepsin B, part of the papain family of proteases, is a widely expressed lysosomal cysteine endopeptidase (24,25). Research studies have suggested that cathepsin B facilitates host cell entry of SARS-CoV by promoting fusion of viral and endosomal membranes (26).

  1. Schmidt, B.L. et al. (2000) J Clin Microbiol 38, 1279-82.
  2. Li, W. et al. (2005) EMBO J 24, 1634-43.
  3. Hoffmann, M. et al. (2020) Cell 181, 271-280.e8.
  4. Lan, J. et al. (2020) Nature 581, 215-20.
  5. Mentzel, S. et al. (1996) J Histochem Cytochem 44, 445-61.
  6. Röhrborn, D. et al. (2015) Front Immunol 6, 386.
  7. Zhong, J. et al. (2015) J Diabetes Res 2015, 606031.
  8. Wang, N. et al. (2013) Cell Res 23, 986-93.
  9. Luan, Y. and Xu, W. (2007) Curr Med Chem 14, 639-47.
  10. Mina-Osorio, P. (2008) Trends Mol Med 14, 361-71.
  11. Biswas, C. et al. (1995) Cancer Res 55, 434-9.
  12. Liao, C.G. et al. (2011) Mol Cell Biol 31, 2591-604.
  13. Sweeny, L. et al. (2012) Exp Cell Res 318, 1788-98.
  14. Lescaille, G. et al. (2012) BMC Cancer 12, 115.
  15. Wang, K. et al. (2020) Signal Transduct Target Ther 5, 283.
  16. Mu, F.T. et al. (1995) J Biol Chem 270, 13503-11.
  17. Christoforidis, S. et al. (1999) Nature 397, 621-5.
  18. Burkard, C. et al. (2014) PLoS Pathog 10, e1004502.
  19. Diamond, M.S. and Farzan, M. (2013) Nat Rev Immunol 13, 46-57.
  20. Brass, A.L. et al. (2009) Cell 139, 1243-54.
  21. Feeley, E.M. et al. (2011) PLoS Pathog 7, e1002337.
  22. Wang, Q. et al. J Struct Biol 146, 44-57.
  23. Wong, H.H. et al. (2015) J Virol 89, 11116-28.
  24. Chan, S.J. et al. (1986) Proc Natl Acad Sci U S A 83, 7721-5.
  25. Fong, D. et al. (1986) Proc Natl Acad Sci U S A 83, 2909-13.
  26. Simmons, G. et al. (2005) Proc Natl Acad Sci U S A 102, 11876-81.

Pathways & Proteins

Explore pathways + proteins related to this product.

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For Research Use Only. Not For Use In Diagnostic Procedures.
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U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
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