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45394
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit #45394

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SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 1
Western blot analysis of extracts from various cell lines using Basigin/EMMPRIN (E1S1V) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 2
Western blot analysis of extracts from PC3 cells, mouse brain, rat brain, and His-tagged Neuropilin-1 recombinant protein(1-644) using Neuropilin-1 (D62C6) Rabbit mAb.
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 3
Western blot analysis of extracts from KARPAS 299, C2C12, and H-4-II-E cells using Furin Antibody (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 4
Immunoprecipitation of furin protein from KARPAS 299 cell extracts. Lane 1 is 10% input, lane 2 is Normal Rabbit IgG #2729, and lane 3 is Furin Antibody. Western blot analysis was performed using Furin Antibody. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody.
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 5
Western blot analysis of extracts from mock-infected Vero-E6 cells (lane 1) and SARS-CoV-2-infected Vero-E6 cells (lane 2), using SARS-CoV-2 Spike Protein (RBD) (E7B3E) Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The antibody detects full-length (uncleaved) SARS-CoV-2 spike protein, and the fragment corresponding to the S1 domain generated by endogenous protease cleavage. SARS-CoV-2 infected Vero-E6 cells courtesy of Dr. Mohsan Saeed, National Emerging Infectious Diseases Laboratories, Boston University.
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 6
Western blot analysis of extracts from 293T cells, mock transfected (lane 1) or transiently transfected with expression constructs encoding Myc/DDK-tagged SARS-CoV-2 Spike (lane 2), Myc/DDK-tagged SARS-CoV Spike (lane 3), or Myc/DDK-tagged MERS-CoV Spike (lane 4), using SARS-CoV-2 Spike Protein (RBD) (E7B3E) Rabbit mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The antibody detects full-length (uncleaved) SARS-CoV-2 spike protein, and the fragment corresponding to the S1 domain generated by endogenous protease cleavage. Proteolytic cleavage of SARS-CoV spike protein (middle panel, lane 3) is not observed in these experimental conditions (transient transfection).
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 7
Western blot analysis of purified SARS-CoV-2 Spike RBD (318-541) Recombinant Protein (8xHis-Tag) #48801 (lane 1), SARS-CoV-2 Spike RBD (multimeric) (319-591) Recombinant Protein (8xHis-Tag) #17862 (lane 2), SARS-CoV-2 Spike (trimeric) (16-1208) Recombinant Protein (8xHis-Tag) #65444 (lane 3), and SARS-CoV-2 Spike S1-NTD (16-316) Recombinant Protein (8xHis-Tag) #88587 (lane 4), using SARS-CoV-2 Spike Protein (RBD) (E7B3E) Rabbit mAb (upper) and His-Tag (D3I1O) XP® Rabbit mAb #12698 (lower). Due to the location of the epitope (surrounding Tyr473 of SARS-CoV-2 spike protein), the antibody detects recombinant proteins corresponding to the full-length ectodomain and the receptor binding domain (RBD) of SARS CoV-2 spike protein, but does not detect recombinant protein corresponding only to the SARS-CoV-2 S1 amino-terminal domain (S1-NTD).
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 8
Confocal immunofluorescent analysis of HCT 116 cells transiently transfected with SARS-CoV-2 spike protein, labeled with SARS-CoV-2 Spike Protein (RBD) (E7B3E) Rabbit mAb (left, green) and co-labeled with DyLight 650 Phalloidin #12956 (right, red), and DAPI #4083 (right, blue).
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 9
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.
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 10
Western blot analysis of extracts from various cell lines using Cathepsin L Antibody (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Expression levels of cathepsin L among cell lines are consistent with expectations based on publicly available bioinformatic databases, confirming specificity of the antibody for cathepsin L.
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 11
Western blot analysis of extracts from 293T cells, mock transfected (lane 1) or transiently transfected with an expression construct encoding Myc/DDK-tagged SARS-CoV-2 spike (lane 2), using Cleaved SARS-CoV-2 Spike Protein (Ser686) Antibody (upper), SARS-CoV-2 Spike Protein (S1-NTD) (E7M5X) Mouse mAb #42172 (upper, middle), Myc-Tag (71D10) Rabbit mAb #2278 (lower, middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower). As shown, the cleavage-specific antibody only detects the residual S2 fragment of SARS-CoV-2 spike protein after cleavage at the S1/S2 junction by endogenous proteases.
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 12
Western blot analysis of extracts from unmodified A549 cells (lane 1) and transgenic A549 cells stably expressing ACE2 protein (lane 2), using ACE2 (E5O6J) XP® Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Transgenic A549 cells were generously provided by Dr. Elena Piskounova, Weill Cornell Medicine.
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 13
Western blot analysis of extracts from various cell lines using ACE2 (E5O6J) XP® Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The absence of detectable ACE2 protein in Hep G2 and HeLa cells is consistent with data from both proteomic and molecular expression profiling studies, confirming specificity of the antibody for ACE2.
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 14
Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma using ACE2 (E5O6J) XP® Rabbit mAb performed on the Leica® BOND Rx.
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 15
Western blot analysis of extracts from 293T cells, mock transfected (lane 1) or transiently transfected with plasmid encoding Myc/DDK-tagged SARS-CoV-2 spike protein (lane 2), using SARS-CoV-2 Spike Protein (S1) (E5S3V) Rabbit mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The antibody detects full-length (uncleaved) SARS-CoV-2 spike protein, and the fragment corresponding to the S1 domain generated by endogenous protease cleavage.
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 16
Western blot analysis of SARS-CoV-2 Spike S1-NTD (16-316) Recombinant Protein (8xHis-Tag) #88587 (lane 1), SARS-CoV-2 Spike (trimeric) (16-1208) Recombinant Protein (8xHis-Tag) #65444 (lane 2), SARS-CoV-2 Spike RBD (318-541) Recombinant Protein (8xHis-Tag) #48801 (lane 3), or SARS-CoV-2 Spike RBD (multimeric) (319-591) Recombinant Protein (8xHis-Tag) #17862 (lane 4), using SARS-CoV-2 Spike Protein (S1) (E5S3V) Rabbit mAb (upper) and His-Tag (D3I1O) XP® Rabbit mAb #12698 (lower). Due to the location of the epitope (within the S1 domain of SARS-CoV-2 spike protein), the antibody detects recombinant proteins corresponding to the full-length ectodomain and the S1-NTD of SARS CoV-2 spike protein but does not detect recombinant protein corresponding only to the receptor binding domain (RBD).
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 17
Confocal immunofluorescent analysis of HCT 116 cells transiently transfected with SARS-CoV-2 spike protein, labeled with SARS-CoV-2 Spike Protein (S1) (E5S3V) Rabbit mAb (left, green) and co-labeled with DyLight 650 Phalloidin #12956 (right, red) and DAPI #4083 (right, blue).
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 18
Western blot analysis of extracts from mock-infected Vero-E6 cells (lane 1), SARS-CoV-2-infected Vero-E6 cells (lane 2), and isolated SARS-CoV-2 virions (lane 3), using SARS-CoV-2 Spike Protein (S1) (E5S3V) Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The antibody detects full-length (uncleaved) SARS-CoV-2 spike protein, and the fragment corresponding to the S1 domain generated by endogenous protease cleavage. SARS-CoV-2 virions and infected Vero-E6 cells courtesy of Dr. Mohsan Saeed, National Emerging Infectious Diseases Laboratories, Boston University.
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 19
Immunohistochemical analysis of paraffin-embedded SARS-CoV-2 positive human placenta (left, positive) and normal human placenta (right, negative) using SARS-CoV-2 Spike Protein (S1) (E5S3V) Rabbit mAb. 
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 20
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).
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 21
Western blot analysis of extracts from 293T cells, mock transfected (lane 1) or transiently transfected with plasmid encoding Myc/DDK-tagged cathepsin L protein (lane 2), using cathepsin L Antibody (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 22
Western blot analysis of extracts from 293T cells, mock transfected (lane 1) or transiently transfected with an expression construct encoding Myc/DDK-tagged SARS-CoV-2 spike (lane 2), mock-infected Vero-E6 cells (lane 3), SARS-CoV-2-infected Vero-E6 cells (lane 4), and SARS-CoV-2 virions (lane 5), using Cleaved SARS-CoV-2 Spike Protein (Ser686) Antibody (upper), DYKDDDDK Tag (D6W5B) Rabbit mAb  #14793 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower). As shown, the cleavage-specific antibody only detects the residual S2 fragment of SARS-CoV-2 spike protein after cleavage at the S1/S2 junction by endogenous proteases.
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 23
Immunohistochemical analysis of paraffin-embedded normal human testis using ACE2 (E5O6J) XP® Rabbit mAb performed on the Leica® BOND Rx.
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 24
Immunohistochemical analysis of paraffin-embedded SARS-CoV-2-infected hACE2 K18 mouse lung using SARS-CoV-2 Spike Protein (S1) (E5S3V) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right). Human ACE2 transgenic mouse lung tissue was generously provided by Dr. Nicholas Crossland and Dr. Florian Douam, National Emerging Infectious Diseases Laboratories, Boston University.
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 25
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Basigin/EMMPRIN (E1S1V) Rabbit mAb.
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 26
Immunoprecipitation of cleaved SARS-CoV-2 spike protein (corresponding to the S2 domain) from extracts of 293T cells transiently transfected with full-length SARS-CoV-2 spike protein. Lane 1 is 10% input, lane 2 is Normal Rabbit IgG #2729, and lane 3 is Cleaved SARS-CoV-2 Spike Protein (Ser686) Antibody. Western blot analysis was performed using Cleaved SARS-CoV-2 Spike Protein (Ser686) Antibody. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody. The SARS-CoV-2 spike protein is cleaved by endogenous proteases at the S1/S2 junction (Ser686) during processing of the mature protein.
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 27
Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma using ACE2 (E5O6J) XP® Rabbit mAb performed on the Leica® BOND Rx.
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 28
Immunohistochemical analysis of paraffin-embedded 293T cell pellet transfected with SARS-CoV-2 spike protein (left-top) and various paraffin-embedded human tissues: colon adenocarcinoma (right-top), prostate adenocarcinoma (right-bottom), and salivary gland small cell carcinoma (left-bottom) using SARS-CoV-2 Spike Protein (S1) (E5S3V) Rabbit mAb. Note the lack of staining in the tissues, all of which were procured prior to 2019, and therefore serve as reliable negative controls.
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 29
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).
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 30
Immunohistochemical analysis of paraffin-embedded human esophageal carcinoma using ACE2 (E5O6J) XP® Rabbit mAb performed on the Leica® BOND Rx.
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 31
Immunohistochemical analysis of paraffin-embedded 293T cell pellet, untransfected (left) or SARS-CoV-2 spike protein transfected (right), using SARS-CoV-2 Spike Protein (S1) (E5S3V) Rabbit mAb.
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 32
Immunohistochemical analysis of paraffin-embedded normal human kidney using ACE2 (E5O6J) XP® Rabbit mAb.
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 33
Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma using ACE2 (E5O6J) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 34
Immunohistochemical analysis of a xenograft derived from transgenic A549 cells stably expressing human ACE2 using ACE2 (E5O6J) XP® Rabbit mAb. Transgenic A549 cells were generously provided by Dr. Elena Piskounova, Weill Cornell Medicine.
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 35
Immunohistochemical analysis of paraffin-embedded brain (left), spleen (middle), and lung (right) from wild type (wt/wt, top) and hACE2 AC70 (tg/wt, bottom) mice (Taconic Biosciences, Inc. model #18222) using ACE2 (E5O6J) XP® Rabbit mAb.
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 36
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using ACE2 (E5O6J) XP® Rabbit mAb.
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 37
Immunohistochemical analysis of paraffin-embedded mouse kidney using ACE2 (E5O6J) XP® Rabbit mAb.
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 38
Immunohistochemical analysis of paraffin-embedded mouse forestomach using ACE2 (E5O6J) XP® Rabbit mAb.
SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit: Image 39
Immunohistochemical analysis of paraffin-embedded IGROV-1 cell pellet (left, positive) or Hep G2 cell pellet (right, negative) using ACE2 (E5O6J) XP® Rabbit mAb.
To Purchase # 45394T
製品番号 サイズ 価格 在庫
45394T
1 Kit  (8 x 20 microliters)

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
SARS-CoV-2 Spike Protein (S1) (E5S3V) Rabbit mAb 99423 20 µl
  • WB
  • IHC
  • IF
Vir 110, 220 Rabbit IgG
SARS-CoV-2 Spike Protein (RBD) (E7B3E) Rabbit mAb 63847 20 µl
  • WB
  • IF
Vir 110, 220 Rabbit IgG
Cleaved SARS-CoV-2 Spike Protein (Ser686) Antibody 84534 20 µl
  • WB
  • IP
Vir 100 Rabbit 
ACE2 (E5O6J) XP®Rabbit mAb 92485 20 µl
  • WB
  • IHC
H M 120-135 Rabbit IgG
Neuropilin-1 (D62C6) Rabbit mAb 3725 20 µl
  • WB
  • IP
H M R 120 Rabbit IgG
Basigin/EMMPRIN (E1S1V) Rabbit mAb 13287 20 µl
  • WB
  • IHC
H 38-58 Rabbit IgG
Furin Antibody 43996 20 µl
  • WB
  • IP
H M R 90 Rabbit 
Cathepsin L Antibody 71298 20 µl
  • WB
H Mk 25-42 Rabbit 
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

The SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit provides an economical means of detecting key viral and host proteins involved in SARS-CoV-2 infection of human host cells. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody in the SARS-CoV-2 Virus-Host Interaction Antibody Sampler Kit detects endogenous levels of its target protein. For viral proteins, endogenous expression is defined as levels of target protein present in host cells following infection with the SARS-CoV-2 virus. SARS-CoV-2 Spike Protein (S1) (E5S3V) Rabbit mAb and SARS-CoV-2 Spike Protein (RBD) (E7B3E) Rabbit mAb detect both the full-length SARS-CoV-2 spike protein and the S1 fragment (containing the receptor-binding domain) generated by furin cleavage. Cleaved SARS-CoV-2 Spike Protein (Ser686) Antibody specifically detects the S2 fragment of SARS-CoV-2 spike protein only after cleavage at the S1/S2 junction. The antibodies targeting SARS-CoV-2 proteins do not detect orthologous spike proteins from SARS or MERS coronaviruses. Neuropilin-1 (D62C6) Rabbit mAb detects endogenous levels of total neuropilin-1 protein, but also detects an 80 kDa protein of unknown identity.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Ala254 of human EMMPRIN protein, Asp201 of human ACE2 protein, and Ser459 of SARS-CoV-2 spike protein, with a recombinant protein corresponding to the S1 domain of the SARS-CoV-2 spike protein, and with a GST-fusion protein corresponding to residues of mouse neuropilin-1 protein. Polyclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to the amino terminus (Ser668) of the SARS-CoV-2 spike protein S2 domain, Pro229 of human cathepsin L, and Ala237 of human furin protein.

Background

The cause of the COVID-19 pandemic is a novel and highly pathogenic coronavirus, termed SARS-CoV-2 (severe acute respiratory syndrome coronavirus-2). SARS-CoV-2 is a member of the Coronaviridae family of viruses (1). The SARS-CoV-2 virion is comprised of four key structural proteins: spike (S), envelope (E), membrane (M), and nucleocapsid (N) (2). Coronavirus spike proteins are class I fusion proteins and harbor an ectodomain, a transmembrane domain, and an intracellular tail (3,4). The highly glycosylated ectodomain projects from the viral envelope surface and facilitates attachment and fusion with the host cell membrane. The ectodomain can be further subdivided into the receptor-binding domain (RBD) S1 and membrane-fusion (S2) subunits, which are produced upon proteolysis by host proteases. S1 and S2 subunits are reassociated after cleavage, assembling into crown-like homotrimers (2,4).

The SARS-CoV-2 spike protein contains a novel tetrabasic "furin cleavage site" (FCS) at the S1/S2 junction. Research studies suggest this site is cleaved by proprotein convertases (e.g., furin) or lysosomal proteases (e.g., cathepsin L) (5,6). S1/S2 cleavage elicits a confirmational change in the spike protein that positions elements of the trimeric RBD in an exposed "up" position, priming it for interaction with host receptor proteins. Cleavage can occur at multiple steps of the viral lifecycle, including during viral packaging, or upon contact of the intact virion with the host cell surface. This novel cleavage event has been suggested to contribute to the high infectivity rate of the SARS-CoV-2 virus (7).

The SARS-CoV-2 virus has been shown to utilize the angiotensin-converting enzyme 2 (ACE2) protein as its primary receptor for cellular entry (8). However, research studies have suggested that other cell surface proteins may serve as receptors or co-receptors for SARS-CoV-2. These include neuropilin-1 (NPN1), a single-pass transmembrane receptor that can function as part of a semaphorin receptor complex, and as a vascular endothelial growth factor (VEGF) receptor (9), and Basigin/EMMPRIN (CD147), a type I integral membrane receptor belonging to the immunoglobulin superfamily (10).
  1. Zhou, P. et al. (2020) Nature 579, 270-273.
  2. Tortorici, M.A. and Veesler, D. (2019) Adv Virus Res 105, 93-116.
  3. Li, F. et al. (2006) J Virol 80, 6794-800.
  4. Li, F. (2016) Annu Rev Virol 3, 237-261.
  5. Coutard, B. et al. (2020) Antiviral Res 176, 104742.
  6. Jaimes, J.A. et al. (2020) iScience 23, 101212.
  7. Hasan, A. et al. (2021) J Biomol Struct Dyn 39, 3025-3033.
  8. Shang, J. et al. (2020) Nature 581, 221-224.
  9. Cantuti-Castelvetri, L. et al. (2020) Science 370, 856-860.
  10. Wang, K. et al. (2020) Signal Transduct Target Ther 5, 283.

Pathways & Proteins

Explore pathways + proteins related to this product.

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