Anti-VPS35 Antibody [11H10]

Mouse Anti-Human VPS35 Monoclonal IgG2b

Catalog No. SMC-606

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Species Reactivity Hu, Ms, Rt
Applications WB IHC ICC/IF FCM IP
SKU: SMC-606 Categories: ,

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SMC-606_VPS35_Antibody_11H10_IHC_Human_Intestinal-Cancer_1.png
Mouse Anti-VPS35 Antibody [11H10] used in Immunoprecipitation (IP) on Human A549 cells (SMC-606)Mouse Anti-VPS35 Antibody [11H10] used in Immunoprecipitation (IP) on Human A549 cells (SMC-606)Mouse Anti-VPS35 Antibody [11H10] used in Western Blot (WB) on Human, Mouse A549, MEF (SMC-606)Mouse Anti-VPS35 Antibody [11H10] used in Immunoprecipitation (IP) on Human A549 cells (SMC-606)Mouse Anti-VPS35 Antibody [11H10] used in Immunoprecipitation (IP) on Mouse embryonic fibroblast (SMC-606)
Product Name VPS35 Antibody
Description

Mouse Anti-Human VPS35 Monoclonal IgG2b
Species Reactivity Human, Mouse, Rat
Applications WB, IP, IHC
Antibody Dilution WB (1:1000); IP (1:200); IHC (1:100); optimal dilutions for assays should be determined by the user.; optimal dilutions for assays should be determined by the user.
Host Species Mouse
Immunogen Species Human
Immunogen Full length recombinant human VSP35
Concentration 1 mg/ml
Conjugates APC, ATTO 390, ATTO 488, ATTO 594, Biotin, FITC, HRP, PerCP, RPE, Unconjugated
Dylight 488
Overview:

  • High fluorescence yield
  • High photostability
  • Less pH-sensitive
  • Excellent batch-to-batch reproducibility
  • Stringently QC tested
  • Molecular weight: 1011 g/mol

Dylight 488 Datasheet

 Dylight 488 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 493 nm

λem = 518 nm

εmax = 7.0×104

Laser = 488 nm

 

APC/Cy7
Overview:

  • High quantum yield
  • Excellent batch-to-batch reproducibility
  • Stringently QC tested

APC-Cy7 Datasheet

 

 ACP-Cy7 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 652 nm

λem = 790 nm

Laser = 594 or 633 nm

 

 

  Dylight 350
Overview:

  • High fluorescence intensity
  • High photostability
  • Less pH-sensitive
  • Excellent solubility in water
  • Stringently QC tested
  • Excellent batch-to-batch reproducibility
  • Molecular weight: 874 g/mol

Dylight 350 Datasheet

 Dylight 350 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 353 nm

λem = 432 nm

εmax = 1.5×104

 

 

  Dylight 405
Overview:

  • High fluorescence intensity
  • High photostability
  • Less pH-sensitive
  • Excellent batch-to-batch reproducibility
  • Stringently QC tested
  • Molecular weight: 793 g/mol

Dylight 405 Datasheet

 Dylight 405 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 400 nm

λem = 420 nm

εmax = 3.0×104

Laser = 405 nm

 

Dylight 594
Overview:

  • High fluorescence yield
  • High photostability
  • Less pH-sensitive
  • Excellent batch-to-batch reproducibility
  • Stringently QC tested
  • Molecular weight: 1078 g/mol

Dylight 594 Datasheet

 Dylight 594 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 593 nm

λem = 618 nm

εmax = 8.0×104

Laser = 526 nm

 

 Dylight 633
Overview:

  • High fluorescence yield
  • High photostability
  • Less pH-sensitive
  • Excellent batch-to-batch reproducibility
  • Stringently QC tested
  • Molecular weight: 1066 g/mol

Dylight 633 Datasheet

 Dylight 633 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 638 nm

λem = 658 nm

εmax = 1.7×105

Laser = 633 nm

 

 PerCP 
Overview:

  • Peridinin-Chlorophyll-Protein Complex
  • Small phycobiliprotein
  • Isolated from red algae
  • Large stokes shift (195 nm)
  • Molecular Weight: 35 kDa
  • Applications: The listed applications provide a general overview of potential uses for conjugated antibodies. However, they do not guarantee that every antibody-conjugate combination has been tested or validated for these specific applications.

PerCP Datasheet

  PerCP Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 482 nm

λem = 677 nm

εmax = 1.96 x 106

Laser = 488 nm

 

 PE/ATTO 594
PE/ATTO 594 is a tandem conjugate, where PE is excited at 535 nm and transfers energy to ATTO 594 via FRET (fluorescence resonance energy transfer), which emits at 627 nm.
Overview:

  • High fluorescence yield
  • High photostability
  • Very hydrophilic
  • Excellent solubility in water
  • Very little aggregation

PE/ATTO 594 Datasheet

 PE-ATTO 594 Fluorophore Conjugate Excitation and Emission Spectra Optical Properties:

λex = 535 nm

λem = 627 nm

Laser = 488 to 561 nm

 

  FITC (Fluorescein)
Overview:

  • Excellent fluorescence quantum yield
  • High rate of photobleaching
  • Good solubility in water
  • Broad emission spectrum
  • pH dependent spectra
  • Molecular formula: C20H12O5
  • Molar mass: 332.3 g/mol
  • Applications: The listed applications provide a general overview of potential uses for conjugated antibodies. However, they do not guarantee that every antibody-conjugate combination has been tested or validated for these specific applications.

FITC-Fluorescent-conjugate

 FITC Fluorescein Fluorophore Excitation and Emission Spectra Optical Properties:

λex = 494 nm

λem = 520 nm

εmax = 7.3×104

Φf = 0.92

τfl = 5.0 ns

Brightness = 67.2

Laser = 488 nm

Filter set = FITC

 

 ATTO 700
Overview:

  • High fluorescence yield
  • Excellent thermal and photostability
  • Quenched by electron donors
  • Very hydrophilic
  • Good solubility in polar solvents
  • Zwitterionic dye
  • Molar Mass: 575 g/mol

ATTO 700 Datasheet

  ATTO 700 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 700 nm

λem = 719 nm

εmax = 1.25×105

Φf = 0.25

τfl = 1.6 ns

Brightness = 31.3

Laser = 676 nm

Filter set = Cy®5.5

 

 ATTO 680
Overview:

  • High fluorescence yield
  • Excellent thermal and photostability
  • Quenched by electron donors
  • Very hydrophilic
  • Good solubility in polar solvents
  • Zwitterionic dye
  • Molar Mass: 631 g/mol

ATTO 680 Datasheet

  ATTO 680 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 680 nm

λem = 700 nm

εmax = 1.25×105

Φf = 0.30

τfl = 1.7 ns

Brightness = 37.5

Laser = 633 – 676 nm

Filter set = Cy®5.5

 

 ATTO 655
Overview:

  • High fluorescence yield
  • High thermal and photostability
  • Excellent ozone resistance
  • Quenched by electron donors
  • Very hydrophilic
  • Good solubility in polar solvents
  • Zwitterionic dye
  • Molar Mass: 634 g/mol

ATTO 655 Datasheet

 ATTO 655 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 663 nm

λem = 684 nm

εmax = 1.25×105

Φf = 0.30

τfl = 1.8 ns

Brightness = 37.5

Laser = 633 – 647 nm

Filter set = Cy®5

 

 ATTO 633
Overview:

  • High fluorescence yield
  • High thermal and photostability
  • Moderately hydrophilic
  • Good solubility in polar solvents
  • Stable at pH 4 – 11
  • Cationic dye, perchlorate salt
  • Molar Mass: 652.2 g/mol

ATTO 633 Datasheet

 ATTO 633 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 629 nm

λem = 657 nm

εmax = 1.3×105

Φf = 0.64

τfl = 3.2 ns

Brightness = 83.2

Laser = 633 nm

Filter set = Cy®5

 

 ATTO 594
Overview:

  • High fluorescence yield
  • High photostability
  • Very hydrophilic
  • Excellent solubility in water
  • Very little aggregation
  • New dye with net charge of -1
  • Molar Mass: 1137 g/mol
  • Applications: The listed applications provide a general overview of potential uses for conjugated antibodies. However, they do not guarantee that every antibody-conjugate combination has been tested or validated for these specific applications.

ATTO 594 Datasheet

  ATTO 594 Fluorophore Excitation and Emission Spectrum Optical Properties:

λex = 601 nm

λem = 627 nm

εmax = 1.2×105

Φf = 0.85

τfl = 3.5 ns

Brightness = 102

Laser = 594 nm

Filter set = Texas Red®

 

 ATTO 565
Overview:

  • High fluorescence yield
  • High thermal and photostability
  • Good solubility in polar solvents
  • Excellent solubility in water
  • Very little aggregation
  • Rhodamine dye derivative
  • Molar Mass: 611 g/mol

ATTO 565 Datasheet

  ATTO 565 Fluorophore Excitation and Emission Spectra Optical Properties:

λex = 563 nm

λem = 592 nm

εmax = 1.2×105

Φf = 0.9

τfl = 3.4 n

Brightness = 10

Laser = 532 nm

Filter set = TRITC

 

  ATTO 488
Overview:

  • High fluorescence yield
  • High photostability
  • Very hydrophilic
  • Excellent solubility in water
  • Very little aggregation
  • New dye with net charge of -1
  • Molar Mass: 804 g/mol
  • Applications: The listed applications provide a general overview of potential uses for conjugated antibodies. However, they do not guarantee that every antibody-conjugate combination has been tested or validated for these specific applications. 

ATTO 488 Datasheet

   ATTO 488 Fluorophore Excitation and Emission Spectra Optical Properties:

λex = 501 nm

λem = 523 nm

εmax = 9.0×104

Φf = 0.80

τfl = 4.1 ns

Brightness = 72

Laser = 488 nm

Filter set = FITC

 

  ATTO 390
Overview:

  • High fluorescence yield
  • Large Stokes-shift (89 nm)
  • Good photostability
  • Moderately hydrophilic
  • Good solubility in polar solvents
  • Coumarin derivate, uncharged
  • Low molar mass: 343.42 g/mol
  • Applications: The listed applications provide a general overview of potential uses for conjugated antibodies. However, they do not guarantee that every antibody-conjugate combination has been tested or validated for these specific applications. 

ATTO 390 Datasheet

 ATTO 390 Fluorescent Dye Excitation and Emission Spectra Optical Properties:

λex = 390 nm

λem = 479 nm

εmax = 2.4×104

Φf = 0.90

τfl = 5.0 ns

Brightness = 21.6

Laser = 365 or 405 nm

 

APC (Allophycocyanin)
Overview:

  • High quantum yield
  • Large phycobiliprotein
  • 6 chromophores per molecule
  • Isolated from red algae
  • Molecular Weight: 105 kDa
  • Applications: The listed applications provide a general overview of potential uses for conjugated antibodies. However, they do not guarantee that every antibody-conjugate combination has been tested or validated for these specific applications.

APC Datasheet

  APC Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 650 nm

λem = 660 nm

εmax = 7.0×105

Φf = 0.68

Brightness = 476

Laser = 594 or 633 nm

Filter set = Cy®5

 

Streptavidin

Properties:

  • Homo-tetrameric protein purified from Streptomyces avidinii which binds four biotin molecules with extremely high affinity
  • Molecular weight: 53 kDa
  • Formula: C10H16N2O3S
  • Applications: Western blot, immunohistochemistry, and ELISA

Streptavidin Datasheet

BiotinBiotin Conjugate Structure

Properties:

  • Binds tetrameric avidin proteins including Streptavidin and neuravidin with very high affinity
  • Molar mass: 244.31 g/mol
  • Formula: C10H16N2O3S
  • Applications: Western blot, immunohistochemistry, and ELISA*

*The listed applications provide a general overview of potential uses for conjugated antibodies. However, they do not guarantee that every antibody-conjugate combination has been tested or validated for these specific applications.

Biotin Datasheet

HRP (Horseradish peroxidase)

Properties:

  • Enzymatic activity is used to amplify weak signals and increase visibility of a target
  • Readily combines with hydrogen peroxide (H2O2) to form HRP-H2O2 complex which can oxidize various hydrogen donors
  • Catalyzes the conversion of:
    • Chromogenic substrates (e.g. TMB, DAB, ABTS) into coloured products
    • Chemiluminescent substrates (e.g. luminol and isoluminol) into light emitting products via enhanced chemiluminescence (ECL)
    • Fluorogenic substrates (e.g. tyramine, homovanillic acid, and 4-hydroxyphenyl acetic acid) into fluorescent products
  • High turnover rate enables rapid generation of a strong signal
  • 44 kDa glycoprotein
  • Extinction coefficient: 100 (403 nm)
  • Applications: Western blot, immunohistochemistry, and ELISA*
    • *The listed applications provide a general overview of potential uses for conjugated antibodies. However, they do not guarantee that every antibody-conjugate combination has been tested or validated for these specific applications.

HRP Datasheet

AP (Alkaline Phosphatase)

Properties:

  • Broad enzymatic activity for phosphate esters of alcohols, amines, pyrophosphate, and phenols
  • Commonly used to dephosphorylate the 5’-termini of DNA and RNA to prevent self-ligation
  • Catalyzes the conversion of:
    • Chromogenic substrates (e.g. pNPP, naphthol AS-TR phosphate, BCIP) into coloured products
    • Fluorogenic substrates (e.g. 4-methylumbelliferyl phosphate) into fluorescent products
  • Molecular weight: 140 kDa
  • Applications: Western blot, immunohistochemistry, and ELISA

AP Datasheet

  R-PE (R-Phycoerythrin)
Overview:

  • Broad excitation spectrum
  • High quantum yield
  • Photostable
  • Member of the phycobiliprotein family
  • Isolated from red algae
  • Excellent solubility in water
  • Molecular Weight: 250 kDa

 

  • Applications: The listed applications provide a general overview of potential uses for conjugated antibodies. However, they do not guarantee that every antibody-conjugate combination has been tested or validated for these specific applications.

R-PE Datasheet

  R-PE Fluorophore Excitation and Emission Spectra Optical Properties:

λex = 565 nm

λem = 575 nm

εmax = 2.0×106

Φf = 0.84

Brightness = 1.68 x 103

Laser = 488 to 561 nm

Filter set = TRITC

 
Field of Use Not for use in humans. Not for use in diagnostics or therapeutics. For in vitro research use only.

Properties

Storage Buffer PBS pH 7.4, 50% glycerol, 0.09% Sodium azide *Storage buffer may change when conjugated
Storage Temperature -20ºC, Conjugated antibodies should be stored according to the product label
Shipping Temperature Blue Ice or 4ºC
Purification Protein G Purified
Clonality Monoclonal
Clone Number 11H10
Isotype IgG2b
Specificity Detects ~92 kDa.
Cite This Product Mouse Anti-Human VPS35 Monoclonal (StressMarq Biosciences, Victoria BC, Cat# SMC-606)
Certificate of Analysis A 1:1000 dilution of SMC-606 was sufficient for detection of VPS35 in 10 µg of SH-SY5Y by ECL immunoblot analysis using Goat Anti-Mouse IgG:HRP as the secondary antibody.

Biological Description

Alternative Names VPS35, VPS35 Retromer Complex Component, Vacuolar Protein Sorting-Associated Protein 35, Vacuolar Protein Sorting 35 Homolog, Vesicle Protein Sorting 35, MEM3, PARK17, FLJ10752, HVPS35, Maternal-Embryonic 3, TCCCTA00141
Research Areas Alzheimer's Disease, Cell Signaling, Golgi Proteins, Membrane Trafficking Proteins, Neurodegeneration, Neuroscience, Parkinson's Disease, Protein Trafficking
Cellular Localization Cytoplasm, Endosome, Lysosome, Membrane, Vesicles
Accession Number NP_060676.2
Gene ID 55737
Swiss Prot Q96QK1
Scientific Background Vacuolar Protein Sorter 35 (VPS35) is a core component of the retromer complex, a critical intracellular trafficking system responsible for the endosome-to-Golgi retrieval of membrane proteins. This pathway is essential for maintaining protein homeostasis, synaptic function, and neuronal survival.

Mutations in VPS35—most notably the D620N variant—have been directly linked to familial and sporadic forms of Parkinson’s disease (PD). These mutations impair retromer function, leading to disrupted recycling of key neuronal receptors and transporters. Dysfunctional VPS35 also contributes to mitochondrial fragmentation, oxidative stress, and impaired autophagy—cellular processes that are central to the pathogenesis of neurodegenerative diseases.

Beyond Parkinson’s, VPS35 dysfunction has been implicated in Alzheimer’s disease and other proteinopathies, where it affects the trafficking of amyloid precursor protein (APP) and tau. Its role in regulating endosomal sorting and mitochondrial dynamics positions VPS35 as a critical node in the cellular pathways that maintain neuronal integrity.

As a result, VPS35 is emerging as a promising therapeutic target in neurodegenerative disease research. Restoring or enhancing retromer function may offer novel strategies to counteract protein misfolding, synaptic loss, and neuroinflammation.
References 1. Vilarino-Guell, C. et al. (2011) Am J Hum Genet 89:162–167
2. Zimprich, A. et al. (2011) Am J Hum Genet 89:168–175
3. Rahman, A.A., Morrison, B.E. (2019) Neurosci 401:1-10.

Product Images

<p>Immunohistochemistry analysis using Mouse Anti-VPS35 Monoclonal Antibody, Clone 11H10 (SMC-606). Tissue: Intestinal Cancer. Species: Human. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody (SMC-606) at 1:100 for Overnight at 4C, then 30 min at 37C. Secondary Antibody: Goat Anti-Mouse IgG (H+L): FITC for 45 min at 37C. Counterstain: DAPI for 3 min at RT. Magnification: 20X.</p>

Immunohistochemistry analysis using Mouse Anti-VPS35 Monoclonal Antibody, Clone 11H10 (SMC-606). Tissue: Intestinal Cancer. Species: Human. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody (SMC-606) at 1:100 for Overnight at 4C, then 30 min at 37C. Secondary Antibody: Goat Anti-Mouse IgG (H+L): FITC for 45 min at 37C. Counterstain: DAPI for 3 min at RT. Magnification: 20X.

<p>Immunoprecipitation analysis using Mouse Anti-VPS35 Monoclonal Antibody, Clone 11H10 (SMC-606). Tissue: A549 cells. Species: Human. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody (SMC-606).</p>

Immunoprecipitation analysis using Mouse Anti-VPS35 Monoclonal Antibody, Clone 11H10 (SMC-606). Tissue: A549 cells. Species: Human. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody (SMC-606).

<p>Immunoprecipitation analysis using Mouse Anti-VPS35 Monoclonal Antibody, Clone 11H10 (SMC-606). Tissue: A549 cells. Species: Human. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody (SMC-606). 500 µL cell culture supernatants were incubated with 10 µL of Protein A/G resin beads for 1 hour at 4⁰C. SMC-606 clone 11H10 depletes VPS35 from the A549 cell extract..</p>

Immunoprecipitation analysis using Mouse Anti-VPS35 Monoclonal Antibody, Clone 11H10 (SMC-606). Tissue: A549 cells. Species: Human. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody (SMC-606). 500 µL cell culture supernatants were incubated with 10 µL of Protein A/G resin beads for 1 hour at 4⁰C. SMC-606 clone 11H10 depletes VPS35 from the A549 cell extract..

<p>Western Blot analysis of Human SH-SY5Y showing detection of VPS35 protein using Mouse Anti-VPS35 Monoclonal Antibody, Clone 11H10 (SMC-606). Lane 1: Molecular Weight Ladder. Lane 2: SH-SY5Y (10 ug). Load: 10 µg. Block: 5% Skim Milk powder in TBST. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody (SMC-606) at 1:1000 for 2 hours at RT with shaking. Secondary Antibody: Goat anti-mouse IgG:HRP at 1:4000 for 1 hour at RT with shaking. Color Development: Chemiluminescent for HRP (Moss) for 5 min in RT.</p>

Western Blot analysis of Human SH-SY5Y showing detection of VPS35 protein using Mouse Anti-VPS35 Monoclonal Antibody, Clone 11H10 (SMC-606). Lane 1: Molecular Weight Ladder. Lane 2: SH-SY5Y (10 ug). Load: 10 µg. Block: 5% Skim Milk powder in TBST. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody (SMC-606) at 1:1000 for 2 hours at RT with shaking. Secondary Antibody: Goat anti-mouse IgG:HRP at 1:4000 for 1 hour at RT with shaking. Color Development: Chemiluminescent for HRP (Moss) for 5 min in RT.

<p>Western Blot analysis of Human, Mouse A549, MEF showing detection of VPS35 protein using Mouse Anti-VPS35 Monoclonal Antibody, Clone 11H10 (SMC-606). Lane 1: Molecular Weight Ladder. Lane 2: VPS35 KO A549 cells. Lane 3: mouse embryonic fibroblast cells.. Load: 8 µg each A549 and MEF. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody (SMC-606) at 1:5 (tissue culture supernatant). Secondary Antibody: Donkey anti-mouse IRDye 800CW at 1:25000 in TBS-T.</p>

Western Blot analysis of Human, Mouse A549, MEF showing detection of VPS35 protein using Mouse Anti-VPS35 Monoclonal Antibody, Clone 11H10 (SMC-606). Lane 1: Molecular Weight Ladder. Lane 2: VPS35 KO A549 cells. Lane 3: mouse embryonic fibroblast cells.. Load: 8 µg each A549 and MEF. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody (SMC-606) at 1:5 (tissue culture supernatant). Secondary Antibody: Donkey anti-mouse IRDye 800CW at 1:25000 in TBS-T.

<p>Immunoprecipitation analysis using Mouse Anti-VPS35 Monoclonal Antibody, Clone 11H10 (SMC-606). Tissue: embryonic fibroblast. Species: Mouse. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody (SMC-606). Three amounts of SMC-606 (3, 1 and 0.3 ug) were non-covalently coupled to 10uL of A/G sepharose beads for 1 hour at 4°C and next incubated with 250ug of MEF lysate for 2 hours at 4°C.</p>

Immunoprecipitation analysis using Mouse Anti-VPS35 Monoclonal Antibody, Clone 11H10 (SMC-606). Tissue: embryonic fibroblast. Species: Mouse. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody (SMC-606). Three amounts of SMC-606 (3, 1 and 0.3 ug) were non-covalently coupled to 10uL of A/G sepharose beads for 1 hour at 4°C and next incubated with 250ug of MEF lysate for 2 hours at 4°C.

<p>Immunoprecipitation analysis using Mouse Anti-VPS35 Monoclonal Antibody, Clone 11H10 (SMC-606). Tissue: A549 cells. Species: Human. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody (SMC-606). Three amounts of SMC-606 (3, 1 and 0.3 ug) were non-covalently coupled to 10uL of A/G sepharose beads for 1 hour at 4°C and next incubated with 250ug of A549 lysate for 2 hours at 4°C.</p>

Immunoprecipitation analysis using Mouse Anti-VPS35 Monoclonal Antibody, Clone 11H10 (SMC-606). Tissue: A549 cells. Species: Human. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody (SMC-606). Three amounts of SMC-606 (3, 1 and 0.3 ug) were non-covalently coupled to 10uL of A/G sepharose beads for 1 hour at 4°C and next incubated with 250ug of A549 lysate for 2 hours at 4°C.

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