Influenza A Matrix Protein antibody | GA2B
Mouse anti Influenza A Matrix Protein antibody, clone GA2B used for the evaluation of influenza matrix protein expression in infected cell lysates by western blotting.
Image caption:
Tyrosine kinase activity is required for efficient uptake of IAV. (A, B, C) A549 cells were treated with genistein (50 μM, [+]) or (D, E) a mixture of several RTK inhibitors (each 10 μM, [+] see experimental procedures) or the solvent control (DMSO,[−]) respectively for 60 min at 37°C or over night prior to infection. (A) Upon genistein treatment, cells were either mock infected or infected with influenza virus A/FPV/Bratislava/79 (H7N7) (FPV; MOI = 100) for further 30 min. Virus particles were visualized in immunofluorescence (IF) microscopy, via a hemagglutinin (HA)-specific rabbit antiserum and an Alexa 594-conjugated chicken anti-rabbit IgG. The nuclei were stained with DAPI. The white arrowheads point to virus particles lining at the cell border. (B, D) After infection with influenza virus A/Puerto-Rico/8/34 (H1N1) PR8 (MOI = 8) for further 30 min, an acidic wash (PBS, pH 1.3, 4°C) was performed before cell-lysis. Internalized virus particles were visualized with a M1 monoclonal antibody in Western-blot (WB) analysis. Equal protein load was verified by ERK2 detection. The relative amount of M1 was quantified (B and D lower panels). Relative M1 densities are expressed as mean ±SD of three independent experiments. (C, E) Cells were infected with FPV or PR8 (MOI = 4) for 8 h; upon the virus-attachment period an acidic wash (PBS-HCl, pH 5.5) was included. Progeny virus titers were determined by standard plaque assays. Data represent mean values of at least three independent experiments ±SD. Statistical significance was assessed by student's t-test (*) p<0.05, (**) p<0.005.
From: Eierhoff T, Hrincius ER, Rescher U, Ludwig S, Ehrhardt C (2010) The Epidermal Growth Factor Receptor (EGFR) Promotes Uptake of Influenza A Viruses (IAV) into Host Cells. PLoS Pathog 6(9): e1001099.
Mouse anti Influenza A Matrix Protein antibody, clone GA2B used for the evaluation of influenza matrix protein expression in infected cell lysates by western blotting.
Image caption:
EGFR is involved in IAV uptake. (A, B) An EGFR expressing construct was transfected into A549 cells. (A) After 24 h cells were infected with PR8 (MOI = 4) for the indicated time points. An acidic wash (PBS, pH 1.3, 4°C) was performed before cell-lysis to remove not internalized virus particles. In WB analysis M1 and ERK2 were detected. The relative amount of M1 was quantified (A, lower panel). Relative M1 densities are expressed as mean ±SD of three independent experiments. (B) Cells were infected with FPV (MOI = 0.5, left panel) or PR8 (MOI = 0.1, right panel) for 8 h; upon the attachment period an acidic wash (PBS-HCl, pH 5.5) was included. Progeny virus titers were determined by standard plaque assays. Data represent mean values of at least three independent experiments ±SD. Statistical significance was assessed by student's t-test (*) p<0.05. In WB analysis EGFR expression level of uninfected cells was detected. (C, D) NIH3T3 wild-type (wt) and NIH3T3 EGFR expressing (HER14) cells were incubated with (C) EGF (30 ng ml−1, 10 min, 37°C) or (D) were infected with PR8 (MOI = 4) for the indicated times. Upon infection an acidic wash (PBS, pH 1.3, 4°C) was performed before cell-lysis. WBs were probed with the indicated antibodies (pEGFR (Y1068) is the autophosphorylation site in the EGFR at tyrosine 1068). (E) NIH3T3 wild-type (wt) and NIH3T3-Her14 (HER14) cells were infected with PR8 and FPV (MOI = 1) for the indicated times. Upon the attachment period an acidic wash (PBS-HCl, pH 5.5) was included. Progeny virus titers were determined by standard plaque assays. Data represent mean values of three independent experiments ±SD.
From: Eierhoff T, Hrincius ER, Rescher U, Ludwig S, Ehrhardt C (2010) The Epidermal Growth Factor Receptor (EGFR) Promotes Uptake of Influenza A Viruses (IAV) into Host Cells. PLoS Pathog 6(9): e1001099.
Mouse anti Influenza A Matrix Protein antibody, clone GA2B used for the evaluation of influenza matrix protein expression in infected cell lysates by western blotting.
Image caption:
EGFR knock-down impairs efficient uptake of IAV into A549 cells. (A) A549 cells were treated with an EGFR-specific antibody (5 μg ml−1) or mouse serum at the indicated times before and during infection with FPV (MOI = 0.5) for 8 h. Progeny virus titers were determined by standard plaque assays; virus yields of serum treated cells were arbitrarily set as 100%. Calculation of virus titers revealed 5.65×105 PFU ml−1 (±1.2×105) for -30 min pre-incubation, 10×105 PFU ml−1 (±1×105) for anti EGFR- and for serum pre-incubation. Statistical significance was assessed by student's t-test (*) p<0.05. (B, C) A549 cells were transfected with a control siRNA or a specific siRNA targeting the EGFR. 48 h post transfection cells were infected with (B) PR8 (MOI = 4) for 30 min or (C) PR8 or FPV (MOI = 4) for 8 h. (C) Upon the attachment period an acidic wash (PBS-HCl, pH 5.5) was included. (D, E) A549 cells were incubated with EGF (100 ng ml−1) 90 min before infection with (D) PR8 (MOI = 4) for 30 min or (E) PR8 or FPV (MOI = 4) for 8 h. (B, D) Before cell-lysis an acidic wash (PBS-HCl, pH 1.3, 4°C) was performed and cell-lysates were analysed in WB. Relative M1 densities are expressed as mean ±SD of three independent experiments. (C, E) Progeny virus yields were determined by standard plaque assays. Data represent mean values of three independent experiments ±SD. Statistical significance was assessed by student's t-test (*) p<0.05. EGFR knock-down was verified by WB before infection (C and E, lower panels). Cells were transiently transfected with an EGFR construct during RNAi mediated knock-down (B) or 24 h prior to EGF induced EGFR degradation (D).
From: Eierhoff T, Hrincius ER, Rescher U, Ludwig S, Ehrhardt C (2010) The Epidermal Growth Factor Receptor (EGFR) Promotes Uptake of Influenza A Viruses (IAV) into Host Cells. PLoS Pathog 6(9): e1001099.
Mouse anti Influenza A Matrix Protein antibody, clone GA2B used for the evaluation of influenza matrix protein expression in infected cell lysates by western blotting.
Image caption:
The RTK c-Met is involved in IAV internalization. (A, B) A549 cells were incubated with a c-Met kinase inhibitor (50 μM) at 37°C over night or (C, D) were transfected with control or c-Met specific siRNA 48 h prior to infection. Internalized influenza virus particles were visualized as described in Figure 1B upon infection with the influenza virus strain PR8 (A) (MOI = 8) or (C) (MOI = 4). Progeny virus titers were determined 8 h upon infection with the influenza A virus strain FPV (B, D) (MOI = 4). Data represent mean values of three independent experiments ±SD. Statistical significance was assessed by student's t-test (*) p<0.05, c-Met knock-down was verified by WB before infection (D, lower panel). (E) A549 cells were transfected either with a control siRNA or with specific siRNAs against c-Met and EGFR, as indicated 48 h prior to infection. Progeny virus titers were determined 8 h upon infection with FPV or PR8 (MOI = 4). Data represent mean values of three independent experiments ±SD. Statistical significance was assessed by student's t-test (*) p<0.05, (**) p<0.005. EGFR and c-Met knock-down was verified by WB before infection (E, lower panels). (F) A549 cells were treated with an inhibitor mix containing the indicated RTK inhibitors (each 10 μM, see experimental procedures) or the solvent control (DMSO) 16 hours prior to infection at 37°C. Subsequently cells were infected with FPV (MOI = 4) for 8 h and progeny virus titers were determined. Data represent mean values of three independent experiments ±SD. Statistical significance was assessed by student's t-test (*) p<0.05.
From: Eierhoff T, Hrincius ER, Rescher U, Ludwig S, Ehrhardt C (2010) The Epidermal Growth Factor Receptor (EGFR) Promotes Uptake of Influenza A Viruses (IAV) into Host Cells. PLoS Pathog 6(9): e1001099.
Mouse anti Influenza A Matrix Protein antibody, clone GA2B used for the evaluation of influenza matrix protein expression in infected cell lysates by western blotting.
Image caption:
Attachment of IAV clusters plasma-membrane lipids. (A, B) A549 cells were infected with FPV (MOI = 100), stimulated with EGF (100 ng ml−1) or left untreated for 1 h at 4°C. Subsequently cells were incubated with the FITC-conjugated Choleratoxin beta subunit (CtxB) (30 μg ml−1) for 1 h at 4°C to visualize ganglioside M1 (GM1). (B) FPV was visualized by detection of HA via an HA-specific rabbit antiserum and an Alexa 594-conjugated chicken anti-rabbit IgG (right panel) or via an H7-HA-specific mouse antiserum and a Texas-Red conjugated goat anti-mouse IgG (middle panel). EGFR was detected by an EGFR-specific mouse antiserum followed by a Texas-Red conjugated goat anti-mouse IgG (left panel) or a Alexa 488-conjugated chicken anti-mouse IgG (right panel). For co-patching analysis CtxB was cross-linked by a rabbit antiserum against CtxB. Cells were examined by confocal laser scanning-microscopy. The colocalization was quantified as described in the experimental procedure section. (C, D, E) A549 cells were incubated with methyl-β-cyclodextrin (MCD) (40 μg ml−1) for 1 h at 37°C and subsequently washed with PBS to withdraw MCD. (C) Cells were stained for GM1 with FITC-conjugated CtxB for 1 h at 4°C and subsequently incubated with rabbit antiserum against CtxB. (D) Cells were infected with PR8 (MOI = 4) for 1 h at 37°C; an acidic wash (PBS-HCl; pH 1.3, 4°C) was performed. In WB analysis M1 and ERK2 were detected. Relative M1 densities are expressed as mean values ±SD of at least three independent experiments. (E) Cells were infected with FPV or PR8 (MOI = 4); upon the attachment period an acidic wash (PBS-HCl, pH 5.5) was performed. Progeny virus titers were determined by plaque assays. Data represent mean values of three independent experiments ±SD. Statistical significance was assessed by student's t-test (*) p<0.05, (**) p<0.005.
From: Eierhoff T, Hrincius ER, Rescher U, Ludwig S, Ehrhardt C (2010) The Epidermal Growth Factor Receptor (EGFR) Promotes Uptake of Influenza A Viruses (IAV) into Host Cells. PLoS Pathog 6(9): e1001099.
Mouse anti Influenza A Matrix Protein antibody, clone GA2B used for the evaluation of influenza matrix protein expression in infected cell lysates by western blotting.
Image caption:
IAV attachment induces EGFR endocytosis. (A) Upon treatment with sialidase (± sialidase; 0.01 units ml−1, 3 h, 37°C), A549 cells were infected with FPV (MOI = 100) or were stimulated with EGF, each for 1 h at 4°C and 30 min at 37°C. An EGFR-specific mouse antiserum and Alexa 488-conjugated chicken anti-mouse IgG as well as a HA-specific rabbit antiserum and an Alexa 594-conjugated chicken anti-rabbit IgG were employed. Cells were examined by IF microscopy. (B, C) A549 cells were infected with FPV (MOI = 100) or (B) incubated with EGF (30 ng ml−1) for 1 h at 4°C upon treatment with sialidase (± sialidase; 0.01 units ml−1, 3 h, 37°C). After a temperature shift cells were further kept at 37°C for the indicated times. (B) Surface resident EGFR was detected by FACS analysis. Fluorescence of uninfected/-stimulated cells was arbitrarily set as 100%. (C) After an acidic wash (PBS-HCl, pH 1.3), cells were permeabilized with saponin (0.2% w/v). Infected cells were assessed by detection of viral HA in FACS analysis. (D, E, F, G) A549 cells were transfected with a control or specific siRNAs targeting caveolin-1 (CAV-1) or clathrin heavy chain (CHC). (D) 48 h post transfection, knock-down was verified by WB using the indicated antibodies. (E, F) upon knock-down cells were (E) infected with FPV or (F) incubated with EGF as described in (B), without sialidase treatment. Cell surface EGFR was detected as mentioned in (B). (G) FPV internalization upon knock-down of the indicated proteins was analysed as described in (C).
From: Eierhoff T, Hrincius ER, Rescher U, Ludwig S, Ehrhardt C (2010) The Epidermal Growth Factor Receptor (EGFR) Promotes Uptake of Influenza A Viruses (IAV) into Host Cells. PLoS Pathog 6(9): e1001099.
Mouse anti Influenza A Matrix Protein antibody, clone GA2B used for the evaluation of influenza matrix protein expression in infected cell lysates by western blotting.
Image caption:
Forced expression of influenza viral proteins does not result in impaired IFNβ-induced STAT1 and STAT2 phosphorylation. HEK293 cells were transfected with 500 ng plasmid DNA for expression of viral NP, M, NS, (A) PA, PB1 and PB2 (C) genes (see Table 1 for accession numbers of viral genes) using L2000 according to manufacturer's instructions. Note that the Pol II constructs in use also give rise to expression of second reading frames in the NS, M and PB1 genes (NS2, M2, PB1-F2). 48 h post transfection cells were stimulated with human IFNβ (500 U/ml) for 15 minutes. Total protein lysates were subjected to Western blot analysis using anti-phospho-STAT1, anti-phospho-STAT2, anti-STAT1 antibodies. Expression of influenza viral proteins was monitored with antibodies against NP, M1, NS1, PA, PB1 or PB2. (E) HEK293 cells were infected with the human influenza virus PR8 (H1N1) (MOI = 5) for the indicated time points and were subsequently stimulated for 15 min with either human IFNβ at a concentration of 100 U/ml. Cell lysates were subjected to Western blots as described. (B, D, F) Quantification of relative pSTAT1 and pSTAT2 band intensities in A, C and E using AIDA software and 2D densitometry (Fuji).
From: Pauli E-K, Schmolke M, Wolff T, Viemann D, Roth J, Bode JG, et al. (2008) Influenza A Virus Inhibits Type I IFN Signaling via NF-κB-Dependent Induction of SOCS-3 Expression.
PLoS Pathog 4(11): e1000196.
Mouse anti Influenza A Matrix Protein antibody, clone GA2B used for the evaluation of influenza matrix protein expression in infected cell lysates by western blotting.
Image caption:
Characterization of influenza H5N1 VLPs. A) A schematic diagram of wild type and mutant H5 HA. The mutant H5 HA is showing a deletion of polybasic amino acids (RRRKK) in the cleavage region of HA (ΔH5 HA). B) Western blot analysis of purified H5N1 VLPs. Lane 1, Influenza H5N1 VLPs. Lane 2, M1 VLPs lacking H5 HA. C) Cleavage of HA in VLPs. H5N1 VLPs were incubated without (−) or with (+) TPCK-treated trypsin (2.0 μg/ml trypsin), resolved on SDS-PAGE, and probed by western blot. D) Negative stain electron microscopy of influenza H5N1 VLPs.
From: Kang S-M, Yoo D-G, Lipatov AS, Song J-M, Davis CT, Quan F-S, et al. (2009) Induction of Long-Term Protective Immune Responses by Influenza H5N1 Virus-Like Particles.
PLoS ONE 4(3): e4667.
Mouse anti Influenza A matrix protein 1 antibody, clone GA2B recognizes an epitope within the influenza A matrix protein 1. In both strains of virus used as immunogen to isolate clone GA2B, the matrix protein 1 is a 252 amino acid, highly conserved viral protein playing a crucial role in replication.
Mouse anti Influenza A matrix protein 1 antibody, clone GA2B can be used in influenza A IFA typing in conjunction with Mouse anti Influenza A matrix protein, clone AA5H.
Mouse anti Influenza A matrix protein 1 antibody, clone GA2B can be used in influenza A IFA typing in conjunction with Mouse anti Influenza A matrix protein, clone AA5H.
Product Details
- Target Species
- Viral
- Product Form
- Purified IgG - liquid
- Preparation
- Purified IgG prepared by affinity chromatography on Protein A from tissue culture supernatant.
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
0.09% Sodium Azide - Immunogen
- Influenza A / Puerto Rico / 8 / 34 (H1N1) and A/Bangkok / 1 / 79 (H3N2) viruses.
- Purity
- >90% IgG content as established by SDS PAGE
- Approx. Protein Concentrations
- IgG concentration 1.0 mg/ml
- Fusion Partners
- Spleen cells from immunised BALB/c mice were fused with cells of the P3 Ag8.653 mouse myeloma cell line.
Storage Information
- Storage
- Store at +4oC or at -20oC if preferred.
This product should be stored undiluted.
Storage in frost free freezers is not recommended. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use. - Shelf Life
- 18 months from date of despatch.
More Information
- UniProt
- P03485 Related reagents
- P03487 Related reagents
- Entrez Gene
- M1 Related reagents
- GO Terms
- GO:0005886 plasma membrane
- GO:0003723 RNA binding
- GO:0005829 cytosol
- GO:0005198 structural molecule activity
- GO:0005654 nucleoplasm
- GO:0019064 viral envelope fusion with host membrane
- GO:0019061 uncoating of virus
- GO:0019065 receptor-mediated endocytosis of virus by host
- GO:0019070 viral genome maturation
- GO:0019072 viral genome packaging
- GO:0019076 release of virus from host
- GO:0019083 viral transcription
- GO:0030666 endocytic vesicle membrane
- GO:0030683 evasion by virus of host immune response
- GO:0031904 endosome lumen
- GO:0042025 host cell nucleus
- GO:0046796 viral genome transport in host cell
- GO:0055036 virion membrane
- GO:0016020 membrane
- Regulatory
- For research purposes only
Applications of Influenza A Matrix Protein antibody
| Application Name | Verified | Min Dilution | Max Dilution |
|---|---|---|---|
| Immunofluorescence | 1/100 | ||
| Immunohistology - Paraffin | |||
| Western Blotting |
Where this antibody has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the antibody for use in their own system using appropriate negative/positive controls.
Copyright © 2018 Bio-Rad Antibodies (formerly AbD Serotec)
Secondary Antibodies Available
Product Specific References
References for Influenza A Matrix Protein antibody
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