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TMEM119 Recombinant Rabbit Monoclonal Antibody [PSH12-98] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA751453

TMEM119 Recombinant Rabbit Monoclonal Antibody [PSH12-98] - BSA and Azide free

Application

  • WB

  • IF-Cell

  • FC

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

TMEM119 Recombinant Rabbit Monoclonal Antibody [PSH12-98] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within mouse TMEM119 aa 113-280.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, FC, IP

Molecular Weight

Predicted band size: 29 kDa

Positive Control

SH-SY5Y cell lysate, Mouse brain tissue lysate, Mouse cerebellum tissue lysate, Rat brain tissue lysate, SH-SY5Y.

Conjugation

unconjugated

Clone Number

PSH12-98

Product Features

Form

Liquid

Concentration

Storage Instructions

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

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IF-Cell

  • 1:100

  • FC

  • 1:1,000

  • IP

  • 1-2μg/sample

Target

Function

Plays an important role in bone formation and normal bone mineralization. Promotes the differentiation of myoblasts into osteoblasts. May induce the commitment and differentiation of myoblasts into osteoblasts through an enhancement of BMP2 production and interaction with the BMP-RUNX2 pathway. Up-regulates the expression of ATF4, a transcription factor which plays a central role in osteoblast differentiation. Essential for normal spermatogenesis and late testicular differentiation (By similarity).

Background References

1. Kenkhuis B et al. Co-expression patterns of microglia markers Iba1, TMEM119 and P2RY12 in Alzheimer\'s disease. Neurobiol Dis. 2022 Jun

2. Sun T et al. TMEM119 facilitates ovarian cancer cell proliferation, invasion, and migration via the PDGFRB/PI3K/AKT signaling pathway. J Transl Med. 2021 Mar

Subcellular Location

Cell membrane, Cytoplasm, Endoplasmic reticulum membrane, Secreted.

UNIPROT

SwissProt: Q4V9L6 Human

SwissProt: Q8R138 Mouse

Entrez Gene: 304581 Rat

Synonyms

OBIF antibody

Osteoblast induction factor antibody

PSEC0199 antibody

Transmembrane protein 119 antibody

UNQ731/PRO1415 antibody

Images

  • Western blot analysis of TMEM119 on different lysates with Rabbit anti-TMEM119 antibody (<a href="/products/HA751453" style="font-weight: bold;text-decoration: underline;">HA751453</a>) at 1/2,000 dilution.<br /><br />Lane 1: SH-SY5Y cell lysate (20 µg/Lane)<br />Lane 2: Mouse brain tissue lysate (40 µg/Lane)<br />Lane 3: Mouse cerebellum tissue lysate (40 µg/Lane)<br />Lane 4: Rat brain tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 29 kDa<br />Observed band size: 35/55 kDa<br /><br />Exposure time: Lane 1: 3 minutes; Lane 2-4: 14 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/HA751453" style="font-weight: bold;text-decoration: underline;">HA751453</a>) at 1/2,000 dilution was used in 5% NFDM/TBST 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 TMEM119 on different lysates with Rabbit anti-TMEM119 antibody (HA751453) at 1/2,000 dilution.

    Lane 1: SH-SY5Y cell lysate (20 µg/Lane)
    Lane 2: Mouse brain tissue lysate (40 µg/Lane)
    Lane 3: Mouse cerebellum tissue lysate (40 µg/Lane)
    Lane 4: Rat brain tissue lysate (40 µg/Lane)

    Predicted band size: 29 kDa
    Observed band size: 35/55 kDa

    Exposure time: Lane 1: 3 minutes; Lane 2-4: 14 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 (HA751453) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of SH-SY5Y cells labeling TMEM119 with Rabbit anti-TMEM119 antibody (<a href="/products/HA751453" style="font-weight: bold;text-decoration: underline;">HA751453</a>) at 1/200 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-TMEM119 antibody (<a href="/products/HA751453" style="font-weight: bold;text-decoration: underline;">HA751453</a>) at 1/200 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 SH-SY5Y cells labeling TMEM119 with Rabbit anti-TMEM119 antibody (HA751453) at 1/200 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-TMEM119 antibody (HA751453) at 1/200 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.

  • Flow cytometric analysis of SH-SY5Y cells labeling TMEM119.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA751453" style="font-weight: bold;text-decoration: underline;">HA751453</a>, 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&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of SH-SY5Y cells labeling TMEM119.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA751453, 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).

  • TMEM119 was immunoprecipitated from 0.2 mg mouse cerebellum tissue lysate with <a href="/products/HA751453" style="font-weight: bold;text-decoration: underline;">HA751453</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA751453" style="font-weight: bold;text-decoration: underline;">HA751453</a> 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.<br /><br />Lane 1: Mouse cerebellum tissue lysate (input)<br />Lane 2: <a href="/products/HA751453" style="font-weight: bold;text-decoration: underline;">HA751453</a> IP in mouse cerebellum tissue lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA751453" style="font-weight: bold;text-decoration: underline;">HA751453</a> in mouse cerebellum tissue lysate<br /><br />Blocking/Dilution buffer: primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>)<br />Exposure time: 3 minutes; ECL: <a href="/products/K1802" style="font-weight: bold;text-decoration: underline;">K1802</a>

    TMEM119 was immunoprecipitated from 0.2 mg mouse cerebellum tissue lysate with HA751453 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751453 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: Mouse cerebellum tissue lysate (input)
    Lane 2: HA751453 IP in mouse cerebellum tissue lysate
    Lane 3: Rabbit IgG instead of HA751453 in mouse cerebellum tissue lysate

    Blocking/Dilution buffer: primary antibody dilution (K1803)
    Exposure time: 3 minutes; ECL: K1802

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