VPS35 | HUABIO HA751603 product information

product summary

company name :

HUABIO

product type :

antibody

product name :

VPS35

catalog :

HA751603

quantity :

100μl

price :

385.00 USD

clonality :

monoclonal

host :

domestic rabbit

conjugate :

nonconjugated

clone name :

PSH16-54

reactivity :

human, mouse, rat, rhesus macaque

application :

western blot, immunoprecipitation, flow cytometry, immunohistochemistry - paraffin section

more info or order :

HUABIO product webpage

image

image 1 :

Western blot analysis of VPS35 on different lysates with Rabbit anti-VPS35 antibody (HA751603) at 1/5,000 dilution. Lane 1: SH-SY5Y cell lysate (20 µg/Lane) Lane 2: A549 cell lysate (20 µg/Lane) Lane 3: U-937 cell lysate (20 µg/Lane) Lane 4: COS-1 cell lysate (20 µg/Lane) Lane 5: Neuro-2a cell lysate (20 µg/Lane) Lane 6: NIH/3T3 cell lysate (20 µg/Lane) Lane 7: C2C12 cell lysate (20 µg/Lane) Lane 8: C6 cell lysate (20 µg/Lane) Lane 9: NR8383 cell lysate (20 µg/Lane) Lane 10: PC-12 cell lysate (20 µg/Lane) Lane 11: Mouse brain tissue lysate (40 µg/Lane) Lane 12: Mouse colon tissue lysate (40 µg/Lane) Lane 13: Rat brain tissue lysate (40 µg/Lane) Lane 14: Rat colon tissue lysate (40 µg/Lane) Predicted band size: 92 kDa Observed band size: 80 kDa Exposure time: 59 seconds; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA751603) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

image 2 :

Western blot analysis of VPS35 on different lysates with Rabbit anti-VPS35 antibody (HA751603) at 1/5,000 dilution. Lane 1: HAP1-parental cell lysate (10 µg/Lane) Lane 2: HAP1-VPS35 KD cell lysate (10 µg/Lane) Predicted band size: 92 kDa Observed band size: 80 kDa Exposure time: 3 minutes; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA751603) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

image 3 :

Immunocytochemistry analysis of SH-SY5Y cells labeling VPS35 with Rabbit anti-VPS35 antibody (HA751603) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-VPS35 antibody (HA751603) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

product information

SKU :

HA751603

Target name :

VPS35

Species reactivity :

Human,Mouse,Rat,Monkey

Applications :

WB,IF-Cell,FC,IHC-P,IP

Conjugate :

Non-conjugated

Immunogen :

Synthetic peptide within human VSP35 aa 760-795.

Uniprot id :

Q96QK1>SwissProt: Q96QK1 Human;SwissProt: Q9EQH3 Mouse;SwissProt: G3V8A5 Rat;SwissProt: I0FFY6 Monkey

Host :

Rabbit

Clone number :

PSH16-54

Isotype :

IgG

Size :

100μl

List Price :

385.00 USD

Storage Buffer :

PBS (pH7.4).

Form :

Liquid

Storage Instruction :

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Purity :

Protein A affinity purified.

Product type :

Recombinant Rabbit monoclonal Antibody

Positive control :

SH-SY5Y cell lysate, A549 cell lysate, U-937 cell lysate, COS-1 cell lysate, Neuro-2a cell lysate, NIH/3T3 cell lysate, C2C12 cell lysate, C6 cell lysate, NR8383 cell lysate, PC-12 cell lysate, Mouse brain tissue lysate, Mouse colon tissue lysate, Rat brain tissue lysate, Rat colon tissue lysate, SH-SY5Y, Neuro-2a, C6, human brain tissue, human colon carcinoma tissue, human kidney tissue, mouse brain tissue, mouse kidney tissue, rat brain tissue, rat kidney tissue.

Molecular wt :

Predicted band size: 92 kDa

Subcellular location :

Cytoplasm, Membrane, Endosome, Early endosome, Late endosome.

Concentration :

1 mg/mL.

Recommended dilutions :

WB: 1:5,000 ;IF-Cell: 1:100 ;IHC-P: 1:200-1:1,000 ;FC: 1:1,000 ;IP: 1-2μg/sample

Advanced Validation :

Knockdown (KD)

Pic img4 :

https://storage.huabio.cn/huabio/productImg/HA751603_4.jpg

Pic legend4 :

Immunocytochemistry analysis of Neuro-2a cells labeling VPS35 with Rabbit anti-VPS35 antibody (HA751603) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-VPS35 antibody (HA751603) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Pic img5 :

https://storage.huabio.cn/huabio/productImg/HA751603_5.jpg

Pic legend5 :

Immunocytochemistry analysis of C6 cells labeling VPS35 with Rabbit anti-VPS35 antibody (HA751603) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-VPS35 antibody (HA751603) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Pic img6 :

https://storage.huabio.cn/huabio/productImg/HA751603_6.jpg

Pic legend6 :

Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-VPS35 antibody (HA751603) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA751603) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Pic img7 :

https://storage.huabio.cn/huabio/productImg/HA751603_7.jpg

Pic legend7 :

Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-VPS35 antibody (HA751603) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA751603) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Pic img8 :

https://storage.huabio.cn/huabio/productImg/HA751603_8.jpg

Pic legend8 :

Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-VPS35 antibody (HA751603) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA751603) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Pic img9 :

https://storage.huabio.cn/huabio/productImg/HA751603_9.jpg

Pic legend9 :

Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-VPS35 antibody (HA751603) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA751603) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Pic img10 :

https://storage.huabio.cn/huabio/productImg/HA751603_10.jpg

Pic legend10 :

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-VPS35 antibody (HA751603) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA751603) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Pic img11 :

https://storage.huabio.cn/huabio/productImg/HA751603_11.jpg

Pic legend11 :

Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-VPS35 antibody (HA751603) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA751603) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Pic img12 :

https://storage.huabio.cn/huabio/productImg/HA751603_12.jpg

Pic legend12 :

Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-VPS35 antibody (HA751603) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA751603) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Pic img13 :

https://storage.huabio.cn/huabio/productImg/HA751603_13.jpg

Pic legend13 :

Flow cytometric analysis of SH-SY5Y cells labeling VPS35. Cells were fixed and permeabilized. Then stained with the primary antibody (HA751603, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Pic img14 :

https://storage.huabio.cn/huabio/productImg/HA751603_14.jpg

Pic legend14 :

Flow cytometric analysis of Neuro-2a cells labeling VPS35. Cells were fixed and permeabilized. Then stained with the primary antibody (HA751603, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Pic img15 :

https://storage.huabio.cn/huabio/productImg/HA751603_15.jpg

Pic legend15 :

Flow cytometric analysis of C6 cells labeling VPS35. Cells were fixed and permeabilized. Then stained with the primary antibody (HA751603, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Pic img16 :

https://storage.huabio.cn/huabio/productImg/HA751603_16.jpg

Pic legend16 :

VPS35 was immunoprecipitated from 0.2 mg A549 cell lysate with HA751603 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751603 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: A549 cell lysate (input) Lane 2: HA751603 IP in A549 cell lysate Lane 3: Rabbit IgG instead of HA751603 in A549 cell lysate Blocking/Dilution buffer: primary antibody dilution (K1803) Exposure time: 3 minutes; ECL: K1801

Pic img17 :

https://storage.huabio.cn/huabio/productImg/HA751603_17.jpg

Pic legend17 :

VPS35 was immunoprecipitated from 0.2 mg NIH/3T3 cell lysate with HA751603 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751603 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: NIH/3T3 cell lysate (input) Lane 2: HA751603 IP in NIH/3T3 cell lysate Lane 3: Rabbit IgG instead of HA751603 in NIH/3T3 cell lysate Blocking/Dilution buffer: primary antibody dilution (K1803) Exposure time: 46 seconds; ECL: K1801

more info or order :

HUABIO product webpage