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Goat Anti-VPS35 / MEM3 Antibody (EB06268)

Code Name Applications Availability Size Tested species Price Grade  
EB06268 Goat Anti-VPS35 / MEM3 Antibody Pep-ELISA, WB, IHC Yes 100µg specific antibody in 200µl Human, Rat $315.00 Elite

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EB06268 1
EB06268 (1µg/ml) staining of Human Brain (Cerebellum) (A) and Rat Brain (B) lysates (35µg protein in RIPA buffer). Detected by chemiluminescence.
EB06268 2
EB06268 (5µg/ml) staining of paraffin embedded Human Prostate. Steamed antigen retrieval with citrate buffer pH 6, AP-staining.
EB06268 3
EB06268 (5µg/ml) staining of paraffin embedded Human Small Intestine. Steamed antigen retrieval with citrate buffer pH 6, AP-staining.
EB06268 4
EB06268 staining of PFA-fixed and saponin permeabilized HEK293 and detected with FITC in confocal microscopy. Data obtained from Aarhus University Denmark. This data is from a previous batch, not on sale.
EB06268 5
EB06268 staining of PFA-fixed and saponin permeabilized SHSY5Y and detected with FITC in confocal microscopy. Data obtained from Dr. M. Schallburg Nielsen, Aarhus University Denmark. This data is from a previous batch, not on sale.
Click here to download PDF datasheet.

Target Protein

Principal Names: VPS35, MEM3, FLJ10752, FLJ13588, FLJ20388, DKFZp434E1211, DKFZp434P1672, vacuolar protein sorting 35 (yeast), maternal-embryonic 3, vacuolar protein sorting 35 homolog (S. cerevisiae), vacuolar protein sorting 35
Official Symbol: VPS35
Accession Number(s): NP_060676.2
Human GeneID(s): 55737
Non-Human GeneID(s): 65114 (mouse)
Important Comments: Note there is a hypothetical protein called similar to vacuolar protein sorting 35 (XP_040192.1), which is virtually identical.

Immunogen

Peptide with sequence C-SPESEGPIYEGLIL, from the C Terminus of the protein sequence according to NP_060676.2.

Please note the peptide is available for sale.

Purification and Storage

Purified from goat serum by ammonium sulphate precipitation followed by antigen affinity chromatography using the immunizing peptide.
Supplied at 0.5 mg/ml in Tris saline, 0.02% sodium azide, pH7.3 with 0.5% bovine serum albumin.
Aliquot and store at -20°C. Minimize freezing and thawing.

Applications Tested

Peptide ELISA: antibody detection limit dilution 1:128000.
Western blot: Approx 85-90kDa band observed in Human Brain (Cerebellum), Heart and Rat Brain lysates (calculated MW of 91.7kDa according to NP_060676.2). Recommended concentration: 1-3µg/ml. Preliminary testing was unsuccessful on Mouse Brain for this particular batch. Primary incubation was 1 hour.
IHC: Paraffin embedded Human Prostate and Small Intestine (Vessel). Recommended concentration: 5µg/ml.

Immunocytochemistry: Vesicular staining of VPS35 (green) in HEK293 and in SHSY5Y. Based on these data, a previous batch of this antibody was endorsed by PabmAbs (http://pabmabs.com/wordpress/?p=1523).

Species Reactivity

Tested: Human, Rat
Expected from sequence similarity: Human, Mouse, Rat, Cow

Specific References

This antibody has been successfully used in IF on Human:
Takatori S, Tatematsu T, Cheng J, Matsumoto J, Akano T, Fujimoto T.
Phosphatidylinositol 3,5-Bisphosphate-Rich Membrane Domains in Endosomes and Lysosomes
Traffic. 2016 Feb;17(2):154-67
PMID: 26563567

This antibody has been successfully used in ICC on CHO cells:
Lee S, Uchida Y, Emoto K, Umeda M, Kuge O, Taguchi T, Arai H.
Impaired retrograde membrane traffic through endosomes in a mutant CHO cell defective in phosphatidylserine synthesis.
Genes Cells. 2012 Aug;17(8):728-36. doi: 10.1111/j.1365-2443.2012.01622.x.
PMID: 22747682

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