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68355
Host Cell Viral Restriction Factor Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Host Cell Viral Restriction Factor Antibody Sampler Kit #68355

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Simple Western™ analysis of HT-29 + hIFN⍺ -1 (10ng/mL, O/N) + poly(I:C )(2.5µg/mL, 7hr) + MG-132 (10µM, 7hr) lysates (0.1 mg/mL) using OAS1 (D1W3A) Rabbit mAb #14498. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.

Simple Western™ analysis of lysates (0.1 mg/mL) from HeLa cells using IFITM3 (D8E8G) XP® Rabbit mAb #59212. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Western blot analysis of extracts from K-562 cells, CD4+ T cells, and CD8+ T cells using IFITM1 Antibody.
Western blot analysis of extracts from various cell lines using TRIM5α (D6Z8L) Rabbit mAb.
Western blot analysis of extracts from HeLa, NCI-H460, and HT-29 cells, untreated (-) or treated with Human Interferon-α1 (hIFN-α1) #8927 (10 ng/ml, 16 hr; +), using OAS1 (D1W3A) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from various cell lines using BST2 (D5V5Z) Rabbit mAb.
Western blot analysis of extracts from various cell lines using RNase L (D4B4J) Rabbit mAb.
Western blot analysis of extracts from C2C12 cells, untreated or thapsigargin-treated, using Phospho-eIF2α (Ser51) (D9G8) XP® Rabbit mAb (upper) or eIF2α Antibody #9722 (lower).
Western blot analysis of extracts from HeLa cells, untreated (-) or treated with Human Interferon-α1 (hIFN-α1) #8927 (10 ng/ml, 16 hr; +), using MX1 (D3W7I) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from various cell lines using IFITM3 (D8E8G) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
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.
Western blot analysis of extracts from THP-1 cells, untreated (-) or treated with calf intestinal phosphatase (CIP) plus λ phosphatase (+), using Phospho-SAMHD1 (Thr592) (D7O2M) (upper) or SAMHD1 Antibody #12361 (lower).
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing full-length human IFITM1 (hIFITM1; +), using IFITM1 Antibody.
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human TRIM5α (hTRIM5α-Myc/DDK), using TRIM5α (D6Z8L) Rabbit mAb.
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing human OAS1 isoform p41 (hOAS1 p41; +), using OAS1 (D1W3A) Rabbit mAb.
Western blot analysis of extracts from 293T cells, untransfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human RNase L (hRNase L-Myc/DDK; +), using RNase L (D4B4J) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma, untreated (left) or λ phosphatase-treated (right), using Phopsho-eIF2α (Ser51) (D9G8) XP® Rabbit mAb.
Immunoprecipitation of MX1 from extracts of HeLa cells treated with Human Interferon-α1 #8927 (10 ng/ml, 16 hr). Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is MX1 (D3W7I) Rabbit mAb. Western blot analysis was performed using MX1 (D3W7I) Rabbit mAb.
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.
Western blot analysis of extracts from Jurkat and THP-1 cells, untreated (-) or treated with TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174 (80 nM, 16 hr; +), using Phospho-SAMHD1 (Thr592) (D7O2M) Rabbit mAb (upper), SAMHD1 Antibody #12361 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).
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).
Immunoprecipitation of TRIM5α from RL cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or TRIM5α (D6Z8L) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using TRIM5α (D6Z8L) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-eIF2α (Ser51) (D9G8) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human ductal carcinoma of the breast using MX1 (D3W7I) Rabbit mAb performed on the Leica® Bond Rx.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using IFITM3 (D8E8G) XP® Rabbit mAb.
Immunoprecipitation of phospho-SAMHD1 (Thr592) from THP-1 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-SAMDHD1 (Thr592) (D7O2M) Rabbit mAb. Western blot analysis was performed using Phospho-SAMDHD1 (Thr592) (D7O2M) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human lymphoma using Phospho-eIF2α (Ser51) (D9G8) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded T84 (left, positive) and HCT116 (right, negative) cell pellets using MX1 (D3W7I) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded normal human lymph node using IFITM3 (D8E8G) XP® Rabbit mAb.
Flow cytometric analysis of THP-1 cells, treated with TPA (80 nM, 16 hr; red) or untreated (green), using Phospho-SAMHD1 (Thr592) (D7O2M) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Immunohistochemical analysis of paraffin-embedded HeLa cell pellets, untreated (left) or treated with Human Interferon-alpha1 (hIFN-alpha1) #8927 (right) using MX1 (D3W7I) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded HeLa (left) and RPMI 8226 (right) cell pellets using IFITM3 (D8E8G) XP® Rabbit mAb.
Flow cytometric analysis of Jurkat cells (blue) and THP-1 cells (green) using Phospho-SAMHD1 (Thr592) (D7O2M) Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using MX1 (D3W7I) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded normal human spleen using IFITM3 (D8E8G) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using MX1 (D3W7I) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Immunohistochemical analysis of paraffin-embedded normal human spleen using MX1 (D3W7I) Rabbit mAb.
To Purchase # 68355
Cat. # Size Qty. Price
68355T
1 Kit  (9 x 20 microliters)

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
MX1 (D3W7I) Rabbit mAb 37849 20 µl
  • WB
  • IP
  • IHC
H 76 Rabbit IgG
OAS1 (D1W3A) Rabbit mAb 14498 20 µl
  • WB
  • IP
H 40, 44 Rabbit IgG
RNase L (D4B4J) Rabbit mAb 27281 20 µl
  • WB
H M 80 Rabbit IgG
IFITM3 (D8E8G) XP® Rabbit mAb 59212 20 µl
  • WB
  • IP
  • IHC
H 15 Rabbit IgG
BST2 (D5V5Z) Rabbit mAb 19277 20 µl
  • WB
H 28-40 Rabbit IgG
TRIM5α (D6Z8L) Rabbit mAb 14326 20 µl
  • WB
  • IP
H Mk 56 Rabbit IgG
Phospho-eIF2α (Ser51) (D9G8) XP® Rabbit mAb 3398 20 µl
  • WB
  • IP
  • IHC
H M R Mk Dm 38 Rabbit IgG
Phospho-SAMHD1 (Thr592) (D7O2M) Rabbit mAb 89930 20 µl
  • WB
  • IP
  • F
H M R 72 Rabbit IgG
IFITM1 Antibody 13126 20 µl
  • WB
H 14 Rabbit 
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

The Host Cell Viral Restriction Factor Antibody Sampler Kit provides an economical means of detecting the expression of various host cell viral restriction factors using phospho-specific and total protein antibodies. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody in the Host Cell Viral Restriction Factor Antibody Sampler Kit detects endogenous levels of its target protein. MX1 (D3W7I) Rabbit mAb recognizes endogenous levels of total MX1 protein. OAS1 (D1W3A) Rabbit mAb recognizes endogenous levels of total OAS1 protein. This antibody cross-reacts with an unidentified protein of 100 kDa in some cell lines. RNase L (D4B4J) Rabbit mAb recognizes endogenous levels of total RNase L protein. IFITM3 (D8E8G) XP® Rabbit mAb recognizes endogenous levels of total IFITM3 protein. This antibody does not cross-react with IFITM1 or IFITM2 proteins. BST2 (D5V5Z) Rabbit mAb recognizes endogenous levels of total BST2 protein. TRIM5α (D6Z8L) Rabbit mAb recognizes endogenous levels of total TRIM5α protein. This antibody does not cross-react with human TRIM5β protein and is not predicted to cross-react with other human TRIM5 isoforms. Phospho-eIF2α (Ser51) (D9G8) XP® Rabbit mAb detects endogenous levels of eIF2α protein only when phosphorylated at Ser51. Phospho-SAMHD1 (Thr592) (D7O2M) Rabbit mAb recognizes endogenous levels of SAMHD1 protein only when phosphorylated at Thr592. IFITM1 Antibody recognizes endogenous levels of total IFITM1 protein. This antibody does not cross-react with IFITM2 or IFITM3 proteins.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu292 of human MX1 protein, Asp90 of human OAS1 protein, Pro717 of human RNase L protein, Val5 of human IFITM3 protein, Val134 of human BST2 protein, Pro395 of human TRIM5α protein, Ser51 of human eIF2α protein, and Thr592 of human SAMHD1 protein. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro20 of human IFITM1 protein. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

Background

Viral restriction factors are proteins produced by host cells that function, in part, to negatively impact various stages of viral life cycles in order to prevent propagation. MX1 (Myxovirus resistance protein 1/MxA) is an interferon-inducible antiviral protein that confers resistance to RNA viruses by blocking transcription of the viral genome (1,2).

2’-5’-oligoadenylate synthetase 1 (OAS1) is an antiviral protein induced by type 1 interferon that plays a key role in the cellular innate immune response (3). The OAS1 enzyme produces a second messenger, 2’-5’-linked oligoadenylate, which binds to RNase L, which then degrades viral and cellular RNA (4). Research studies indicate that the OAS1 system inhibits protein synthesis and induces apoptosis in virally infected cells, which limits viral infection (5).

Interferon-induced transmembrane protein (IFITM) family members, IFITM1 and IFITM3, appear to function as viral restriction factors by preventing fusion of viral and host membranes (6,7).

BST2 (CD317, Tetherin, HM1.24) is a type II transmembrane glycoprotein functioning as a major mediator of the innate immune defense against the dissemination of enveloped viruses by tethering viron on the cell surface, preventing viral release (8).

TRIM5α blocks viral infection by interacting with the incoming viral capsid and promoting its premature disassembly (9).

PKR-induced phosphorylation of the eukaryotic initiation factor 2 (eIF2) α subunit at Ser51 is a well-documented mechanism to downregulate protein synthesis upon viral infection (10).

SAM domain and HD domain-containing protein 1 (SAMHD1) prevents autoimmunity and HIV infection by hydrolyzing intracellular deoxynucleoside triphosphates (dNTPs), thereby limiting inappropriate immune activation by self nucleic acid and inhibiting reverse transcription of the HIV genome (11). Phosphorylation of SAMHD1 at Thr592 by cyclin A2/CDK1 was identified as a regulatory mechanism that controls SAMHD1 activity. SAMHD1 is phosphorylated in proliferating cells, which inhibits its ability to block HIV infection (12).

Pathways

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Limited Uses

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For Research Use Only. Not for Use in Diagnostic Procedures.
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