Anti-VPS35 Antibody [7E4]

Name: VPS35 Antibody
SKU: SMC-602
Availability: InStock

Mouse Anti-Human VPS35 Monoclonal IgG2a

Catalog No. SMC-602

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Species Reactivity	Hu, Ms, Rt
Applications	WB IHC ICC/IF FCM IP

SKU: SMC-602 Categories: Antibodies, Monoclonal

SMC-602_VPS35_Antibody_7E4_ICC-IF_Human_A549-cells_1.png

Mouse Anti-VPS35 Antibody [7E4] used in Immunoprecipitation (IP) on Human, Mouse MEF, A549 cells (SMC-602)

Product Name VPS35 Antibody

Description

Mouse Anti-Human VPS35 Monoclonal IgG2a

Species Reactivity Human, Mouse, Rat

Applications WB, ICC/IF, IP

Antibody Dilution WB (1:1000); ICC/IF (1:200); IP (1:200); 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:

λ_ex = 493 nm

λ_em = 518 nm

ε_max = 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:

λ_ex = 652 nm

λ_em = 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:

λ_ex = 353 nm

λ_em = 432 nm

ε_max = 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:

λ_ex = 400 nm

λ_em = 420 nm

ε_max = 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:

λ_ex = 593 nm

λ_em = 618 nm

ε_max = 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:

λ_ex = 638 nm

λ_em = 658 nm

ε_max = 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

PerCP Datasheet

PerCP Fluorophore Absorption and Emission Spectrum

Optical Properties:

λ_ex = 482 nm

λ_em = 677 nm

ε_max = 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:

λ_ex = 535 nm

λ_em = 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

FITC-Fluorescent-conjugate

FITC Fluorescein Fluorophore Excitation and Emission Spectra

Optical Properties:

λ_ex = 494 nm

λ_em = 520 nm

ε_max = 7.3×10⁴

Φ_f = 0.92

τ_fl = 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:

λ_ex = 700 nm

λ_em = 719 nm

ε_max = 1.25×10⁵

Φ_f = 0.25

τ_fl = 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:

λ_ex = 680 nm

λ_em = 700 nm

ε_max = 1.25×10⁵

Φ_f = 0.30

τ_fl = 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:

λ_ex = 663 nm

λ_em = 684 nm

ε_max = 1.25×10⁵

Φ_f = 0.30

τ_fl = 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:

λ_ex = 629 nm

λ_em = 657 nm

ε_max = 1.3×10⁵

Φ_f = 0.64

τ_fl = 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

ATTO 594 Datasheet

ATTO 594 Fluorophore Excitation and Emission Spectrum

Optical Properties:

λ_ex = 601 nm

λ_em = 627 nm

ε_max = 1.2×10⁵

Φ_f = 0.85

τ_fl = 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:

λ_ex = 563 nm

λ_em = 592 nm

ε_max = 1.2×10⁵

Φ_f = 0.9

τ_fl = 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

ATTO 488 Datasheet

ATTO 488 Fluorophore Excitation and Emission Spectra

Optical Properties:

λ_ex = 501 nm

λ_em = 523 nm

ε_max = 9.0×10⁴

Φ_f = 0.80

τ_fl = 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

ATTO 390 Datasheet

ATTO 390 Fluorescent Dye Excitation and Emission Spectra

Optical Properties:

λ_ex = 390 nm

λ_em = 479 nm

ε_max = 2.4×10⁴

Φ_f = 0.90

τ_fl = 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

APC Datasheet

APC Fluorophore Absorption and Emission Spectrum

Optical Properties:

λ_ex = 650 nm

λ_em = 660 nm

ε_max = 7.0×10⁵

Φ_f = 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

Biotin

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

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

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

R-PE Datasheet

R-PE Fluorophore Excitation and Emission Spectra

Optical Properties:

λ_ex = 565 nm

λ_em = 575 nm

ε_max = 2.0×10⁶

Φ_f = 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	7E4
Isotype	IgG2a
Specificity	Detects ~92 kDa.
Cite This Product	Mouse Anti-Human VPS35 Monoclonal (StressMarq Biosciences, Victoria BC, Cat# SMC-602)
Certificate of Analysis	A 1:1000 dilution of SMC-602 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	Vacuolar protein sorting-associated protein 35, MEM3, PARK17, VPS35 retromer complex component, maternal-embryonic 3, vesicle protein sortin 35, TCCCTA00141, FLJ10752
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 component of the retromer complex, which is essential for endosome-to-Golgi retrieval of membrane proteins. VPS35 mutations such as D620N have been linked to Parkinson’s Disease (PD) (1,2) and affect retromer function, protein homeostasis, and mitochondria (3).
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>Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-VPS35 Monoclonal Antibody, Clone 7E4 (SMC-602). Tissue: A549 cells. Species: Human. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody (SMC-602) at 1:5 (tissue culture supernatant). Secondary Antibody: Donkey anti-mouse: Alexa Fluor 594 at 1:4000 in 0.2% BSA PBS. Counterstain: DAPI. Localization: Vesicles. A) VPS35 KO A549 cells B) WT A549 cells. Courtesy of: Dario Alessi Lab, University of Dundee.</p>

Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-VPS35 Monoclonal Antibody, Clone 7E4 (SMC-602). Tissue: A549 cells. Species: Human. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody (SMC-602) at 1:5 (tissue culture supernatant). Secondary Antibody: Donkey anti-mouse: Alexa Fluor 594 at 1:4000 in 0.2% BSA PBS. Counterstain: DAPI. Localization: Vesicles. A) VPS35 KO A549 cells B) WT A549 cells. Courtesy of: Dario Alessi Lab, University of Dundee.

<p>Western Blot analysis of Human, Mouse A549, MEF showing detection of VPS35 protein using Mouse Anti-VPS35 Monoclonal Antibody, Clone 7E4 (SMC-602). 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-602) 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 7E4 (SMC-602). 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-602) 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 SH-SY5Y lysates showing detection of 91.7 kDa VPS35 protein using Mouse Anti-VPS35 Monoclonal Antibody, Clone 7E4 (SMC-602). Lane 1: Molecular Weight Ladder. Lane 2: SH-SY5Y. Load: 10 μg. Block: 5% Skim Milk powder in TBST. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody (SMC-602) 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. Predicted/Observed Size: 91.7 kDa.</p>

Western Blot analysis of Human SH-SY5Y lysates showing detection of 91.7 kDa VPS35 protein using Mouse Anti-VPS35 Monoclonal Antibody, Clone 7E4 (SMC-602). Lane 1: Molecular Weight Ladder. Lane 2: SH-SY5Y. Load: 10 μg. Block: 5% Skim Milk powder in TBST. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody (SMC-602) 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. Predicted/Observed Size: 91.7 kDa.

<p>Immunoprecipitation analysis using Mouse Anti-VPS35 Monoclonal Antibody, Clone 7E4 (SMC-602). Tissue: A549 cells. Species: Human. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody (SMC-602). 500 µL cell culture supernatants were incubated with 10 µL of Protein A/G resin beads for 1 hour at 4⁰C. SMC-602 clone 7E4 depletes virtually all of the VPS35 from the A549 cell extract..</p>

Immunoprecipitation analysis using Mouse Anti-VPS35 Monoclonal Antibody, Clone 7E4 (SMC-602). Tissue: A549 cells. Species: Human. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody (SMC-602). 500 µL cell culture supernatants were incubated with 10 µL of Protein A/G resin beads for 1 hour at 4⁰C. SMC-602 clone 7E4 depletes virtually all of the VPS35 from the A549 cell extract..

Immunoprecipitation analysis using Mouse Anti-VPS35 Monoclonal Antibody, Clone 7E4 (SMC-602). Tissue: A549 cells. Species: Human. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody (SMC-602).

<p>Immunoprecipitation analysis using Mouse Anti-VPS35 Monoclonal Antibody, Clone 7E4 (SMC-602). Tissue: embryonic fibroblast. Species: Mouse. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody (SMC-602). Three amounts of SMC-602 (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 7E4 (SMC-602). Tissue: embryonic fibroblast. Species: Mouse. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody (SMC-602). Three amounts of SMC-602 (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.

Immunoprecipitation analysis using Mouse Anti-VPS35 Monoclonal Antibody, Clone 7E4 (SMC-602). Tissue: A549 cells. Species: Human. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody (SMC-602). Three amounts of SMC-602 (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 7E4 (SMC-602). Tissue: MEF, A549 cells. Species: Human, Mouse. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody (SMC-602) at 1:5 (tissue culture supernatant). 10 ug antibody were coupled to 10 uL A/G resin beads either covalently (with DMP) or non-covalently (1 hour at 4 degrees). The antibody immunoprecipitates VPS35 in mouse and human cells effectively when covalently coupled to the beads..</p>

Immunoprecipitation analysis using Mouse Anti-VPS35 Monoclonal Antibody, Clone 7E4 (SMC-602). Tissue: MEF, A549 cells. Species: Human, Mouse. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody (SMC-602) at 1:5 (tissue culture supernatant). 10 ug antibody were coupled to 10 uL A/G resin beads either covalently (with DMP) or non-covalently (1 hour at 4 degrees). The antibody immunoprecipitates VPS35 in mouse and human cells effectively when covalently coupled to the beads..

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Size
Conjugate	Clear