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VPS35 Recombinant Rabbit Monoclonal Antibody [JB33-82]

Usd: 385

Specification

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Catalog# ET7107-01

VPS35 Recombinant Rabbit Monoclonal Antibody [JB33-82]

Application

  • WB

  • IHC-P

  • IF-Cell

Reactivity

  • Human

  • Mouse

  • Rat

  • Green monkey

Predicted reactivity

  • Cynomolgus monkey

  • Pig

Predicted species support after-sales service
Conjugation

  • unconjugated

Overview

Product Name

VPS35 Recombinant Rabbit Monoclonal Antibody [JB33-82]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human VPS35 aa 80-220 / 796.

Species Reactivity

Human, Mouse, Rat, Green monkey (Predicted: Cynomolgus monkey, Pig)

Predicted species support after-sales service

Validated Applications

WB, IHC-P, IF-Cell

Molecular Weight

Predicted band size: 92 kDa

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, human colon cancer tissue, human kidney tissue, human placenta tissue, mouse colon tissue, NIH/3T3.

Conjugation

unconjugated

Clone Number

JB33-82

RRID

AB_3070718

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000

  • IHC-P

  • 1:100-1:500

  • IF-Cell

  • 1:100

Target

Function

Vacuolar sorting proteins (VPSs) are required for proper trafficking of endocytic and biosynthetic proteins to the vacuole and play an important role in the budding process of cells. VPS35 (vacuolar protein sorting 35), also known as MEM3, is the 796 amino acid human homolog of the S. cerevisiae Vps35 protein. Localized to the cytoplasm and to the peripheral membrane, VPS35 is an essential component of the retromer complex which is involved in retrieval of lysosomal enzyme receptors from endosomes to the trans-Golgi network. VPS35 is expressed ubiquitously with highest expression in heart, placenta, brain, testis, kidney, colon, ovary and spleen. In addition to its crucial role in the retromer complex, VPS35 is part of a subcomplex that is required to regulate transcytosis of the polymeric immunoglobulin receptor from the basolateral to the apical surface of epithelial cells and hepatocytes.

Background References

1. Arighi C N et al. Role of the mammalian retromer in sorting of the cation-independent mannose 6-phosphate receptor. J Cell Biol 165:123-133 (2004) .

2. Verges M et al. The mammalian retromer regulates transcytosis of the polymeric immunoglobulin receptor. Nat Cell Biol 6:763-769 (2004).

Sequence Similarity

Belongs to the VPS35 family.

Tissue Specificity

Ubiquitous. Highly expressed in heart, brain, placenta, skeletal muscle, spleen, thymus, testis, ovary, small intestine, kidney and colon.

Subcellular Location

Cytoplasm. Membrane. Endosome.

UNIPROT

SwissProt: Q96QK1 Human

SwissProt: Q9EQH3 Mouse

Entrez Gene: 25479 Rat

Synonyms

DKFZp434E1211 antibody

DKFZp434P1672 antibody

FLJ10752 antibody

FLJ13588 antibody

FLJ20388 antibody

hVPS35 antibody

Maternal embryonic 3 antibody

Maternal-embryonic 3 antibody

MEM 3 antibody

MEM3 antibody

Expand

Images

  • Western blot analysis of VPS35 on different lysates with Rabbit anti-VPS35 antibody (<a href="/products/ET7107-01" style="font-weight: bold;text-decoration: underline;">ET7107-01</a>) at 1/5,000 dilution.<br /><br />Lane 1: SH-SY5Y cell lysate (20 µg/Lane)<br />Lane 2: A549 cell lysate (20 µg/Lane)<br />Lane 3: U-937 cell lysate (20 µg/Lane)<br />Lane 4: COS-1 cell lysate (20 µg/Lane)<br />Lane 5: Neuro-2a cell lysate (20 µg/Lane)<br />Lane 6: NIH/3T3 cell lysate (20 µg/Lane)<br />Lane 7: C2C12 cell lysate (20 µg/Lane)<br />Lane 8: C6 cell lysate (20 µg/Lane)<br />Lane 9: NR8383 cell lysate (20 µg/Lane)<br />Lane 10: PC-12 cell lysate (20 µg/Lane)<br />Lane 11: Mouse brain tissue lysate (20 µg/Lane)<br />Lane 12: Mouse colon tissue lysate (20 µg/Lane)<br />Lane 13: Rat brain tissue lysate (20 µg/Lane)<br />Lane 14: Rat colon tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 92 kDa<br />Observed band size: 80 kDa<br /><br />Exposure time: 20 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET7107-01" style="font-weight: bold;text-decoration: underline;">ET7107-01</a>) at 1/5,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of VPS35 on different lysates with Rabbit anti-VPS35 antibody (ET7107-01) 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 (20 µg/Lane)
    Lane 12: Mouse colon tissue lysate (20 µg/Lane)
    Lane 13: Rat brain tissue lysate (20 µg/Lane)
    Lane 14: Rat colon tissue lysate (20 µg/Lane)

    Predicted band size: 92 kDa
    Observed band size: 80 kDa

    Exposure time: 20 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 (ET7107-01) 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.

  • Immunohistochemical analysis of paraffin-embedded human colon cancer tissue using anti-VPS35 antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded human colon cancer tissue using anti-VPS35 antibody. Counter stained with hematoxylin.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-VPS35 antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-VPS35 antibody. Counter stained with hematoxylin.

  • Immunohistochemical analysis of paraffin-embedded human placenta tissue using anti-VPS35 antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded human placenta tissue using anti-VPS35 antibody. Counter stained with hematoxylin.

  • Immunohistochemical analysis of paraffin-embedded mouse colon tissue using anti-VPS35 antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded mouse colon tissue using anti-VPS35 antibody. Counter stained with hematoxylin.

  • Immunocytochemistry analysis of NIH/3T3 cells labeling VPS35 with Rabbit anti-VPS35 antibody (<a href="/products/ET7107-01" style="font-weight: bold;text-decoration: underline;">ET7107-01</a>) at 1/100 dilution.<br /><br />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 (<a href="/products/ET7107-01" style="font-weight: bold;text-decoration: underline;">ET7107-01</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of NIH/3T3 cells labeling VPS35 with Rabbit anti-VPS35 antibody (ET7107-01) 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 (ET7107-01) 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.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of VPS35 on different lysates with Rabbit anti-VPS35 antibody (<a href="/products/ET7107-01" style="font-weight: bold;text-decoration: underline;">ET7107-01</a>) at 1/2,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-VPS35 KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 92 kDa<br />Observed band size: 80 kDa<br /><br />Exposure time: 20 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET7107-01" style="font-weight: bold;text-decoration: underline;">ET7107-01</a>) at 1/2,000 dilution was used in <a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a> at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of VPS35 on different lysates with Rabbit anti-VPS35 antibody (ET7107-01) at 1/2,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-VPS35 KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 92 kDa
    Observed band size: 80 kDa

    Exposure time: 20 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 (ET7107-01) at 1/2,000 dilution was used in 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.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"