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    extracellular-matrix-protein-1-antibody-epr22411-279-ab253185.pdf

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Immunology Immunoglobulins Receptors
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)

  • Datasheet
  • SDS
  • Certificate of Compliance
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)
  • Western blot - Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)
  • Western blot - Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)
  • Immunoprecipitation - Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)
  • Immunoprecipitation - Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)
  • Flow Cytometry (Intracellular) - Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)
  • Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22411-279] to Extracellular matrix protein 1
  • Suitable for: Flow Cyt (Intra), WB, IP, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat

Conjugates logo Related conjugates and formulations

Carrier Free

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Overview

  • Product name

    Anti-Extracellular matrix protein 1 antibody [EPR22411-279]
    See all Extracellular matrix protein 1 primary antibodies
  • Description

    Rabbit monoclonal [EPR22411-279] to Extracellular matrix protein 1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IP, IHC-Pmore details
    Unsuitable for: ICC/IF
  • Species reactivity

    Reacts with: Mouse, Rat
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: 4T1, LLC1, Hepa1-6 whole cell lysate. Mouse and rat skin and liver lysates. Wild-type mouse liver lysate. Flow Cyt (intra): 4T1 cells. IP: Mouse liver and lung whole cell lysate.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine)
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR22411-279
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Immunoglobulins
    • Receptors
    • Immunology
    • Immunoglobulins
    • Heavy Chain
    • IgA
    • Immunology
    • Immunoglobulins
    • Heavy Chain
    • IgM
    • Tags & Cell Markers
    • Cell Type Markers
    • Tumor Associated

Associated products

  • Alternative Versions

    • Anti-Extracellular matrix protein 1 antibody [EPR22411-279] - BSA and Azide free (ab267392)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Related Products

    • Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab253185 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt (Intra)
1/500.
WB
1/1000. Detects a band of approximately 48, 85 kDa (predicted molecular weight: 63 kDa).
IP
1/30.
IHC-P
Use at an assay dependent concentration.
Notes
Flow Cyt (Intra)
1/500.
WB
1/1000. Detects a band of approximately 48, 85 kDa (predicted molecular weight: 63 kDa).
IP
1/30.
IHC-P
Use at an assay dependent concentration.
Application notes
Is unsuitable for ICC/IF.

Target

  • Function

    Involved in endochondral bone formation as negative regulator of bone mineralization. Stimulates the proliferation of endothelial cells and promotes angiogenesis. Inhibits MMP9 proteolytic activity.
  • Tissue specificity

    Expressed in breast cancer tissues. Little or no expression observed in normal breast tissues. Expressed in skin; wide expression is observed throughout the dermis with minimal expression in the epidermis.
  • Involvement in disease

    Defects in ECM1 are the cause of lipoid proteinosis (LiP) [MIM:247100]; also known as lipoid proteinosis of Urbach and Wiethe or hyalinosis cutis et mucosae. LiP is a rare autosomal recessive disorder characterized by generalized thickening of skin, mucosae and certain viscera. Classical features include beaded eyelid papules and laryngeal infiltration leading to hoarseness. Histologically, there is widespread deposition of hyaline material and disruption/reduplication of basement membrane.
  • Cellular localization

    Secreted > extracellular space > extracellular matrix.
  • Target information above from: UniProt accession Q16610 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 13601 Mouse
    • Entrez Gene: 116662 Rat
    • SwissProt: Q61508 Mouse
    • SwissProt: Q62894 Rat
    • Unigene: 3433 Mouse
    • Unigene: 97792 Rat
    • Alternative names

      • ECM 1 antibody
      • Ecm1 antibody
      • ECM1_HUMAN antibody
      • Extracellular matrix protein 1 antibody
      • Secretory component p85 antibody
      • URBWD antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)

      Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling Extracellular matrix protein 1 with ab253185 at 1/5000 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). The section was incubated with ab253185 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

       Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer)

       Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)

      Immunohistochemical analysis of paraffin-embedded Mouse live tissue labeling Extracellular matrix protein 1 with ab253185 at 1/5000 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). The section was incubated with ab253185 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer)

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)

      Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Extracellular matrix protein 1 with ab253185 at 1/5000 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). The section was incubated with ab253185 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer)

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

    • Western blot - Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)
      Western blot - Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)
      All lanes : Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185) at 1/1000 dilution

      Lane 1 : Wild-type mouse liver lysate
      Lane 2 : ECM1 knockout mouse liver lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 63 kDa
      Observed band size: 48,85 kDa why is the actual band size different from the predicted?



      The wild-type and ECM1 knockout mouse liver lysates were kindly provided by an anonymous collaborator.

      ab253185 was shown to specifically react with Extracellular matrix protein 1 in wild-type mouse liver as signal was lost in ECM1 knockout liver. Wild-type and ECM1 knockout samples were subjected to SDS-PAGE. ab253185 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/50,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique.

      Exposure time 7.75 secs.

      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)
      Western blot - Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)
      All lanes : Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185) at 1/1000 dilution

      Lane 1 : 4T1 (mouse mammary gland carcinoma epithelial cell), whole cell lysate
      Lane 2 : LLC1 (mouse lung carcinoma), whole cell lysate
      Lane 3 : Hepa1-6 (mouse hepatoma epithelial cell), whole cell lysate
      Lane 4 : Mouse skin lysate
      Lane 5 : Mouse liver lysate
      Lane 6 : Rat skin lysate
      Lane 7 : Rat liver lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 63 kDa
      Observed band size: 48,85 kDa why is the actual band size different from the predicted?



      Mouse ECM1 has two isoforms: A long isoform reported to be 85 kDa, and the shorter form 48 kDa.

      The molecular weight observed is consistent with what has been described in the literature (PMID: 7608209; PMID: 23202415).

      Exposure times: Lanes 1-2: 7.75 seconds; Lane 3: 1 second; Lanes 4-7: 10 seconds.

      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunoprecipitation - Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)
      Immunoprecipitation - Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)

      Extracellular matrix protein 1 was immunoprecipitated from 0.35 mg mouse lung lysate with ab253185 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab253185 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

      Lane 1: Mouse lung lysate 10µg.

      Lane 2: ab253185 IP in mouse lung lysate.

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab253185 in mouse lung lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 30 seconds.

       

    • Immunoprecipitation - Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)
      Immunoprecipitation - Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)

      Extracellular matrix protein 1 was immunoprecipitated from 0.35 mg mouse liver lysate with ab253185 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab253185 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

      Lane 1: Mouse liver lysate 10µg.

      Lane 2: ab253185 IP in mouse liver lysate.

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab253185 in mouse liver lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 3 mins.

       

    • Flow Cytometry (Intracellular) - Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)
      Flow Cytometry (Intracellular) - Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)

      Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized 4T1 (mouse mammary gland carcinoma epithelial cell) cells labeling Extracellular matrix protein 1 with ab253185 at 1/500 (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. 

       

       

    • Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)
      Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (0)

    Publishing research using ab253185? Please let us know so that we can cite the reference in this datasheet.

    ab253185 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Extracellular matrix protein 1 antibody [EPR22411-279]

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Pig Tissue sections (Kidney)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citric acid from Vector (H3300)
    Permeabilization
    No
    Specification
    Kidney
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
    Fixative
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    DR. Dong Zhou

    Verified customer

    Submitted Dec 01 2022

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