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TMED9 Recombinant Mouse Monoclonal Antibody [A9A8-R]

Usd: 350

Specification

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Catalog# HA601268

TMED9 Recombinant Mouse Monoclonal Antibody [A9A8-R]

Application

  • WB

  • IF-Cell

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

Overview

Product Name

TMED9 Recombinant Mouse Monoclonal Antibody [A9A8-R]

Antibody Type

Recombinant Mouse Monoclonal Antibody

Immunogen

Recombinant protein within Human TMED9 aa 31-220 / 235.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P

Molecular Weight

Predicted band size: 27 kDa

Positive Control

HeLa cell lysate, THP-1 cell lysate, A549 cell lysate, MCF7 cell lysate, HepG2 cell lysate, PC-12 cell lysate, NIH/3T3 cell lysate, mouse liver tissue lysate, rat liver tissue lysate, HepG2, human liver tissue, mouse liver tissue, rat liver tissue.

Conjugation

unconjugated

Clone Number

A9A8-R

RRID

AB_3096693

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG1

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IF-Cell

  • 1:100

  • IHC-P

  • 1:2,000

Target

Function

P24 protein family is a group of transmembrane proteins that are major components of COPI and COPII-coated vesicles. The family is also known as EMP24/GP25L/p24 family and TMP21-like proteins. The latter naming was after transmembrane emp24 domain-containing protein 10 that was found in the human brain. It was claimed to block the beta-amyloid peptide, which is implicated in the pathogenesis of Alzheimer's disease. p24 family proteins localize to the major organelles of the early secretory pathway: the endoplasmic reticulum and the Golgi apparatus, where they seem to be involved in trafficking between the two compartments. In yeast, all p24 family proteins can be removed, causing only a mild phenotype. However, in mammals at least some p24 proteins are essential for survival, e.g. removal of p24δ1 is lethal in mice. p24 family members have been implicated in the biogenesis of COPI and COPII-coated vesicles, transporting membrane-bound proteins through the secretory system, and forming the structure of the endoplasmic reticulum and Golgi.

Background References

1. Li S et al. A new type of ERGIC-ERES membrane contact mediated by TMED9 and SEC12 is required for autophagosome biogenesis. Cell Res. 2022 Feb

2. Puri C et al. TMED9-SEC12, an important "contact" for autophagy. Cell Res. 2022 Feb

Subcellular Location

Endoplasmic reticulum membrane, Golgi apparatus, Endoplasmic reticulum-Golgi intermediate compartment membrane.

UNIPROT

SwissProt: Q9BVK6 Human

SwissProt: Q99KF1 Mouse

SwissProt: Q5I0E7 Rat

Synonyms

Transmembrane emp24 domain-containing protein 9

GMP25

Glycoprotein 25L2

p24 family protein alpha-2 (p24alpha2)

p25

TMED9

GP25L2

Images

  • Western blot analysis of TMED9 on different lysates with Mouse anti-TMED9 antibody (<a href="/products/HA601268" style="font-weight: bold;text-decoration: underline;">HA601268</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: THP-1 cell lysate (20 µg/Lane)<br />Lane 3: A549 cell lysate (20 µg/Lane)<br />Lane 4: MCF7 cell lysate (20 µg/Lane)<br />Lane 5: HepG2 cell lysate (20 µg/Lane)<br />Lane 6: PC-12 cell lysate (20 µg/Lane)<br />Lane 7: NIH/3T3 cell lysate (20 µg/Lane)<br />Lane 8: Mouse liver tissue lysate (40 µg/Lane)<br />Lane 9: Rat liver tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 27 kDa<br />Observed band size: 24 kDa<br /><br />Exposure time: 43 seconds;<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/HA601268" style="font-weight: bold;text-decoration: underline;">HA601268</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Anti-Mouse IgG for IP Nano-secondary antibody (<a href="/products/NBI02H" style="font-weight: bold;text-decoration: underline;">NBI02H</a>) at 1/5,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of TMED9 on different lysates with Mouse anti-TMED9 antibody (HA601268) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: THP-1 cell lysate (20 µg/Lane)
    Lane 3: A549 cell lysate (20 µg/Lane)
    Lane 4: MCF7 cell lysate (20 µg/Lane)
    Lane 5: HepG2 cell lysate (20 µg/Lane)
    Lane 6: PC-12 cell lysate (20 µg/Lane)
    Lane 7: NIH/3T3 cell lysate (20 µg/Lane)
    Lane 8: Mouse liver tissue lysate (40 µg/Lane)
    Lane 9: Rat liver tissue lysate (40 µg/Lane)

    Predicted band size: 27 kDa
    Observed band size: 24 kDa

    Exposure time: 43 seconds;

    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 (HA601268) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Anti-Mouse IgG for IP Nano-secondary antibody (NBI02H) at 1/5,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of HepG2 cells labeling TMED9 with Mouse anti-TMED9 antibody (<a href="/products/HA601268" style="font-weight: bold;text-decoration: underline;">HA601268</a>) at 1/100 dilution.<br /><br />Cells were fixed in 100% precooled methanol for 5 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 Mouse anti-TMED9 antibody (<a href="/products/HA601268" style="font-weight: bold;text-decoration: underline;">HA601268</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</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.

    Immunocytochemistry analysis of HepG2 cells labeling TMED9 with Mouse anti-TMED9 antibody (HA601268) at 1/100 dilution.

    Cells were fixed in 100% precooled methanol for 5 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 Mouse anti-TMED9 antibody (HA601268) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue with Mouse anti-TMED9 antibody (<a href="/products/HA601268" style="font-weight: bold;text-decoration: underline;">HA601268</a>) at 1/2,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA601268" style="font-weight: bold;text-decoration: underline;">HA601268</a>) at 1/2,000 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.

    Immunohistochemical analysis of paraffin-embedded human liver tissue with Mouse anti-TMED9 antibody (HA601268) at 1/2,000 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 (HA601268) at 1/2,000 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.

  • Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Mouse anti-TMED9 antibody (<a href="/products/HA601268" style="font-weight: bold;text-decoration: underline;">HA601268</a>) at 1/2,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA601268" style="font-weight: bold;text-decoration: underline;">HA601268</a>) at 1/2,000 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.

    Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Mouse anti-TMED9 antibody (HA601268) at 1/2,000 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 (HA601268) at 1/2,000 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.

  • Immunohistochemical analysis of paraffin-embedded rat liver tissue with Mouse anti-TMED9 antibody (<a href="/products/HA601268" style="font-weight: bold;text-decoration: underline;">HA601268</a>) at 1/2,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA601268" style="font-weight: bold;text-decoration: underline;">HA601268</a>) at 1/2,000 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.

    Immunohistochemical analysis of paraffin-embedded rat liver tissue with Mouse anti-TMED9 antibody (HA601268) at 1/2,000 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 (HA601268) at 1/2,000 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.

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