VPS35 Antibody

Novus Biologicals | Catalog # NB100-1397

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat, Canine, Chicken, Chinese Hamster

Cited:

Human, Mouse

Predicted:

Bovine (100%). Backed by our 100% Guarantee.

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Peptide ELISA, Immunocytochemistry/ Immunofluorescence

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry/ Immunofluorescence, IF/IHC, WB, IHC

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all VPS35 Primary Antibodies »

Product Specifications

Immunogen

Peptide with sequence C-SPESEGPIYEGLIL corresponding to C-Terminus according to NP_060676.2.

Reactivity Notes

Predicted cross-reactivity based on sequence identity: Bovine. It has been reported in the literature to react with Chinese hamster ovary cells (PMID: 22747682). Mouse reactivity reported in scientific literature (PMID: 28252385). Canine, chicken and rat reactivity reported from verified customer reviews.

Specificity

Note there is a hypothetical protein called similar to vacuolar protein sorting 35 (XP_040192.1), which is virtually identical.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Theoretical MW

91 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for VPS35 Antibody

Western Blot: VPS35 Antibody [NB100-1397]

Western Blot: VPS35 Antibody [NB100-1397]

Western Blot: VPS35 Antibody [NB100-1397] - Staining of HepG2 (A) and HEK293 (B) cell lysate (35 ug protein in RIPA buffer). Antibody at 0.03 ug/mL. Detected by chemiluminescence.
Immunocytochemistry/ Immunofluorescence: VPS35 Antibody [NB100-1397]

Immunocytochemistry/ Immunofluorescence: VPS35 Antibody [NB100-1397]

Immunocytochemistry/Immunofluorescence: VPS35 Antibody [NB100-1397] - Immunofluorescence analysis of paraformaldehyde fixed U2OS cells, permeabilized with 0.15% Triton. Primary incubation 1hr (10ug/ml) followed by Alexa Fluor 488 secondary antibody (2ug/ml), showing cytoplasmic/vesicle staining. The nuclear stain is DAPI (blue). Negative control: Unimmunized goat IgG (10ug/ml) followed by Alexa Fluor 488 secondary antibody (2ug/ml).
Immunohistochemistry-Paraffin: VPS35 Antibody [NB100-1397]

Immunohistochemistry-Paraffin: VPS35 Antibody [NB100-1397]

Immunohistochemistry-Paraffin: VPS35 Antibody [NB100-1397] - Negative Control showing staining of paraffin embedded Human Prostate, with no primary antibody.
Western Blot: VPS35 Antibody [NB100-1397]

Western Blot: VPS35 Antibody [NB100-1397]

Western Blot: VPS35 Antibody [NB100-1397] - 10 ug cell lysate per lane of various cell lines as indicated, immunoblotted with vps35 antibody at 1:2000 at 4C for 20 hrs. Secondary antibody donkey anti-goat HRP at 1:5000 for 1 hr at room temp. WB image submitted by a verified customer review.
Western Blot: VPS35 Antibody [NB100-1397]

Western Blot: VPS35 Antibody [NB100-1397]

Western Blot: VPS35 Antibody [NB100-1397] - HEK293T cell lysate. WB image submitted by a verified customer review.
Western Blot: VPS35 Antibody [NB100-1397]

Western Blot: VPS35 Antibody [NB100-1397]

Western Blot: VPS35 Antibody [NB100-1397] - Staining of Human (A) Mouse (B) and Rat (C) Brain lysate (35 ug protein in RIPA buffer). antibody at 0.03 ug/mL. Detected by chemiluminescence.
Immunocytochemistry/ Immunofluorescence: VPS35 Antibody [NB100-1397]

Immunocytochemistry/ Immunofluorescence: VPS35 Antibody [NB100-1397]

Immunocytochemistry/Immunofluorescence: VPS35 Antibody [NB100-1397] - Confocal microscopy image of human RPE-1 cells fixed with PFA, permeablilized with 0.3% Triton X-100,then immunostained with vps35 antibody at 1:2000 overnight in 1% BSA at 4 degrees. ICC image submitted by a verified customer review.
Immunocytochemistry/ Immunofluorescence: VPS35 Antibody [NB100-1397]

Immunocytochemistry/ Immunofluorescence: VPS35 Antibody [NB100-1397]

Immunocytochemistry/Immunofluorescence: VPS35 Antibody [NB100-1397] - Immunofluorescence analysis of paraformaldehyde fixed HEK293 cells, permeabilized with 0.15% Triton. Primary incubation 1hr (10ug/ml) followed by Alexa Fluor 488 secondary antibody (2ug/ml), showing cytoplasmic/vesicle staining. The nuclear stain is DAPI (blue). Negative control: Unimmunized goat IgG (10ug/ml) followed by Alexa Fluor 488 secondary antibody (2ug/ml).
Immunohistochemistry-Paraffin: VPS35 Antibody [NB100-1397]

Immunohistochemistry-Paraffin: VPS35 Antibody [NB100-1397]

Immunohistochemistry-Paraffin: VPS35 Antibody [NB100-1397] - Staining of paraffin embedded Human Prostate. Antibody at 8 ug/mL. Heat induced antigen retrieval with citrate buffer pH 6, HRP-staining.
VPS35 Antibody

Western Blot: VPS35 Antibody [NB100-1397] -

Retromer Vps35 or SNX1/2 depletion altered EGF-stimulated EGFR degradation. (A) Serum-starved cells were treated with 100 ug/mL of EGF for different time points. At indicated time points, cells were harvested, and equal amounts of protein samples were used for immunoblotting with EGFR antibody. Representative blots from at least three independent experiments are shown. (B) EGFR levels at each time point were normalized to the EGFR level at 0 min for each cell line. Graph represents the difference of EGFR levels during EGF stimulation (means +/- SEM). Two-tailed student’s t-test indicated the difference, * p < 0.05, HeLa control vs. Vps35 KO (120 min); * p < 0.05, HeLa control vs. Vps35 KO (240 min); ** p < 0.01, HeLa control vs. SNX1/2 dKO (240 min). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36359754), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
VPS35 Antibody

Western Blot: VPS35 Antibody [NB100-1397] -

Depletion of retromer or retromer-associated SNXs decreases EGFR protein levels. (A) Steady-state HeLa control, Vps35 KO, SNX1/2 dKO, SNX3 KO, and SNX27 KO cell monolayers grown in complete-DMEM media on coverslips were fixed in ice-cold methanol, stained with EGFR antibody, and count-stained with DAPI. Images were captured on a Leica SP8 DMi8 confocal microscope using a 60× glycerol objective (Scale Bar, 10 um). (B) Equal amounts of protein samples from HeLa control, Vps35 KO, SNX1/2 dKO, SNX3 KO, and SNX27 KO cells were subjected to SDS-PAGE and immunoblotting with the antibodies against EGFR, SNX1, SNX2, SNX3, SNX27, and Vps35, whereas tubulin served as a loading control. Representative blots from at least three independent experiments are shown. (C) The fold differences for EGFR protein levels are presented (means +/- SEM). Two-tailed student’s t-test indicated the difference between HeLa control and HeLa KO cells, ** p < 0.01, HeLa control vs. Vps35 KO; *** p < 0.001, HeLa control vs. SNX1/2 dKO; *** p < 0.001, HeLa control vs. SNX3 KO; *** p < 0.001, HeLa control vs. SNX27 KO. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36359754), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for VPS35 Antibody

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

10 ug/mL

Immunohistochemistry

8 ug/mL

Immunohistochemistry-Paraffin

8 ug/mL

Peptide ELISA

Detection limit 1:128000

Western Blot

0.03 - 0.1 ug/mL
Application Notes
Western blot: Approx 90 kDa band observed in Human Brain (Cerebellum) and Mouse Brain lysates (calculated MW of 91.7 kDa according to NP_060676.2).

Reviewed Applications

Read 5 reviews rated 4.8 using NB100-1397 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

Tris saline (20 mM Tris pH 7.3, 150 mM NaCl), 0.5% BSA

Preservative

0.02% Sodium Azide

Concentration

0.5 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C. Avoid freeze-thaw cycles.

Background: VPS35

Chemokine receptor D6 expression has been documented in human skin, small intestine, large intestine, appendix, lymph node, vessels, tonsil, spleen, as well as from rat brain tissues. ESTs have been isolated from cancerous human lung, uterus, cervix, placenta, blood, and from normal human placenta libraries. Caution: G protein-coupled receptor D6 was previously called CCR10 and/or CCR9. CCR9 and C-C chemokine receptor 9 referred to both chemokine receptor D6 and the unrelated receptor GPR-9-6.

Long Name

VPS35 retromer complex component

Alternate Names

MEM3, PARK17

Entrez Gene IDs

55737 (Human); 65114 (Mouse)

Gene Symbol

VPS35

UniProt

NP_060676.2 (Human)

Additional VPS35 Products

Product Documents for VPS35 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Download SDS

Product Specific Notices for VPS35 Antibody

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Customer Reviews for VPS35 Antibody (5)

4.8 out of 5
5 Customer Ratings
5 Stars
80%
4 Stars
20%
3 Stars
0%
2 Stars
0%
1 Stars
0%

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Customer Images

Showing  1 - 5 的 5 reviews Showing All
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  • VPS35 Antibody
    Name: jerome cattin
    Application: Immunofluorescence
    Sample Tested: INS-1
    Species: Rat
    Verified Customer | Posted 10/29/2020
    20%FCS to block, 0.5% Triton, secondary was Anti-Goat-647
    VPS35 Antibody NB100-1397
  • VPS35 Antibody
    Name: jerome cattin
    Application: Immunocytochemistry
    Sample Tested: hela cell
    Species: Human
    Verified Customer | Posted 05/28/2020
    VPS35 Antibody NB100-1397
  • VPS35 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: HEK293T cell lysate
    Species: Human
    Verified Customer | Posted 05/28/2020
    VPS35 Antibody NB100-1397
  • VPS35 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: RPE-1, DF-1, MDCK canine kidney epithelial cell line and SH-SY5Y whole cell lysates
    Species: Human, Chicken and Canine
    Verified Customer | Posted 01/07/2020
    10ug of various cell lines indicated immunbloted with vps35 antibody 1:2000
    samples prepared in 1.5x lammeli buffer, 10ug of protein loaded per lane transferred onto nitrocellulose at 100V for 1.5 hours 4 degrees. blotting coditions membrane blocked in 5% milk in PBS 0.1% tween 20 (PBST) for 1 hour primary ab incubated 1:2000 in 5% milk in PBS at 4 degrees for 20 hours washed 3X with PBST secondary donkey anti goat HPR 1:5000 for 1 hour room temp washed 2x PBST, 1X PBS blot developed using ECL prime reagent (GE healthcare )
    VPS35 Antibody NB100-1397
  • VPS35 Antibody
    Name: Anonymous
    Application: Immunocytochemistry
    Sample Tested: RPE-1 cells and RPE-1
    Species: Human
    Verified Customer | Posted 01/07/2020
    confocal RPE-1 cells fixed with PFA perm 0.3% tx100 then immunostained with vps35 ab 1:2000 overnight in 1% BSA at 4 degrees.
    VPS35 Antibody NB100-1397

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