VPS35 Polyclonal antibody

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  WB analysis of HEK-293 using 10236-1-AP

  WB result of VPS35 antibody (10236-1-AP; 1:10000; incubated at room temperature for 1.5 hours) with sh-Control and sh-VPS35 transfected HEK-293 cells.

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  WB analysis of HAP1 using 10236-1-AP

  HAP1 (WT and VPS35 KO) lysates prepared with RIPA buffer, 20 μg of protein loaded. 10236-1-AP incubated at 1:500 at 4°C overnight in 5% milk in TBST. Ponceau stained transfers shown for each blot. These data were performed and provided by YCharOS in partnership with Proteintech. YCharOS is an open science company with a mission to characterise all commercial antibodies to improve scientific reproducibility and transparency.

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  WB analysis using 10236-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 10236-1-AP (VPS35 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis using 10236-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 10236-1-AP (VPS35 antibody) at dilution of 1:600 incubated at room temperature for 1.5 hours.

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  IP experiment of HEK-293 using 10236-1-AP

  IP result of anti-VPS35 (IP:10236-1-AP, 4ug; Detection:10236-1-AP 1:2000) with HEK-293 cells lysate 1800 ug.

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  IHC staining of mouse kidney using 10236-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse kidney tissue slide using 10236-1-AP (VPS35 antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse kidney using 10236-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse kidney tissue slide using 10236-1-AP (VPS35 antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HepG2 using 10236-1-AP

  Immunofluorescent analysis of (-20°C Ethanol) fixed HepG2 cells using VPS35 antibody (10236-1-AP) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).

• Featured Product
• KD/KO Validated

VPS35 Polyclonal Antibody for WB, IHC, IF/ICC, IP, ELISA

Cat No. 10236-1-AP

Publications(15)

Reviews (2)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat and More (1)

Applications

WB, IHC, IF/ICC, IP, CoIP, ELISA

Synonyms

hVPS35, Maternal embryonic 3, Maternal-embryonic 3, MEM3, TCCCTA00141

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Tested Applications

• Positive WB detected in: HepG2 cells, A549 cells, HAP1 cells, HEK-293 cells, mouse kidney tissue, rat kidney tissue
• Positive IP detected in: HEK-293 cells
• Positive IHC detected in: mouse kidney tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HepG2 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 2 publications below
• WB: See 14 publications below
• IHC: See 1 publications below
• IF: See 6 publications below
• IP: See 1 publications below
• CoIP: See 2 publications below

Product Information

10236-1-AP targets VPS35 in WB, IHC, IF/ICC, IP, CoIP, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat, monkey
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: VPS35 fusion protein Ag0340
• Full Name: vacuolar protein sorting 35 homolog (S. cerevisiae)
• Calculated Molecular Weight: 92 kDa
• Observed Molecular Weight: 80-92 kDa
• GenBank Accession Number: BC002414
• Gene Symbol: VPS35
• Gene ID (NCBI): 55737
• RRID: AB_2215216
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• UNIPROT ID: Q96QK1
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

VPS35 protein belongs to a group of vacuolar protein sorting (VPS) proteins, which ensure the proper delivery of organelle-specific proteins in eukaryotic cells. VPS35 is the core of a multimeric complex, termed the retromer complex, which is involved in retrograde transport of proteins from endosomes to the trans-Golgi network. Vps35 serves as the core of the multimeric complex by binding directly to Vps26 and Vps29 and SNX1. Northern blot analyses in 16 tissues showed that one transcript of Vps35 with a size of 3.6 kb was highly expressed in brain, heart, testis, ovary, small intestine, spleen, skeletal muscle, and placenta and expressed at moderate or low levels in other tissues. Another transcript of Vps35, a message of 3.0 kb, was also expressed with proportionally lower levels than the 3.6-kb transcript in all the tissues except that the 3.0-kb transcript was not detected in brain. Human Vps35 is mapped at 16q13-q21.

Publications

Species	Application	Title
human	WB	Nat Commun; A genome-wide CRISPR screen identifies host factors that regulate SARS-CoV-2 entry.; Authors - Yunkai Zhu
rat	WB,IF,CoIP	Cell Death Differ; Sorting nexin 3 induces heart failure via promoting retromer-dependent nuclear trafficking of STAT3.; Authors - Jing Lu
human	IF	Proc Natl Acad Sci U S A; SNX27 suppresses SARS-CoV-2 infection by inhibiting viral lysosome/late endosome entry.; Authors - Bo Yang
	WB	Hum Mol Genet; VPS35 pathogenic mutations confer no dominant toxicity but partial loss of function in Drosophila and genetically interact with parkin.; Authors - Bilal R Malik
monkey	WB,IF	Am J Pathol; Dynein Dysfunction Reproduces Age-Dependent Retromer Deficiency: Concomitant Disruption of Retrograde Trafficking Is Required for Alteration in β-Amyloid Precursor Protein Metabolism.; Authors - Nobuyuki Kimura
mouse	WB	J Neurochem; Retromer and Rab2-dependent trafficking mediate PS1 degradation by proteasomes in endocytic disturbance.; Authors - Naoya Ueda ; KD Validated

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.

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FH

Xin (Verified Customer) (10-10-2022)

A very good antibody to detect endogenous VPS35 in mouse brain lysate.

• Applications: Western Blot
• Primary Antibody Dilution: 1:2000
• Cell Tissue Type: mouse brain lysate

VPS35 Antibody Western Blot validation (1:2000 dilution) in mouse brain lysate (Cat no:10236-1-AP)

FH

stephen (Verified Customer) (09-07-2019)

Oc-2 cells fixed with 4% PFA Perm. by 0.3% tx100 for 5 min blocked with 1% BSA in 1XPBS for 2 hours vps35 antibody incubated 1:300 overnight at 4 degrees in 1%BSA in 1x PBS. Co-stained with DAPI (blue) and Ph647 (red) to visualize DNA and F-actin respectively.

• Applications: Immunofluorescence,
• Primary Antibody Dilution: 1:300
• Cell Tissue Type: mouse OC2 cells

VPS35 Antibody Immunofluorescence, validation (1:300 dilution) in mouse OC2 cells (Cat no:10236-1-AP)