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RecombinantRabMAb

Recombinant Anti-Entactin/NID antibody [EPR22414-125] (ab254325)

Reviews (1) Submit a question References (1)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Entactin/NID antibody [EPR22414-125] (ab254325)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Entactin/NID antibody [EPR22414-125] (ab254325)
  • Immunohistochemistry (Frozen sections) - Anti-Entactin/NID antibody [EPR22414-125] (ab254325)
  • Immunohistochemistry (Frozen sections) - Anti-Entactin/NID antibody [EPR22414-125] (ab254325)
  • Western blot - Anti-Entactin/NID antibody [EPR22414-125] (ab254325)
  • Western blot - Anti-Entactin/NID antibody [EPR22414-125] (ab254325)
  • Western blot - Anti-Entactin/NID antibody [EPR22414-125] (ab254325)
  • Anti-Entactin/NID antibody [EPR22414-125] (ab254325)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22414-125] to Entactin/NID
  • Suitable for: WB, IHC-P, IHC-Fr
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-Entactin/NID antibody [EPR22414-125]
    See all Entactin/NID primary antibodies

  • Description

    Rabbit monoclonal [EPR22414-125] to Entactin/NID

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, IHC-Frmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • Positive control

    • WB: Recombinant human Entactin/NID protein (aa29 -1114); Human kidney, skeletal muscle, lung and placenta tissue lysate; Mouse heart, lung, skeletal muscle, kidney tissue lysate. IHC-P: Mouse cardiac tissue and rat cardiac muscle tissue. IHC-Fr: Mouse and rat heart tissue.

  • 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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab254325 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
WB
1/1000. Predicted molecular weight: 136 kDa.
IHC-P (1)
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

IHC recommended for Mouse and Rat only.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
IHC-Fr
1/500.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)
Notes
WB
1/1000. Predicted molecular weight: 136 kDa.
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

IHC recommended for Mouse and Rat only.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
IHC-Fr
1/500.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)
Application notes
Is unsuitable for IP.

Target

  • Function

    Sulfated glycoprotein widely distributed in basement membranes and tightly associated with laminin. Also binds to collagen IV and perlecan. It probably has a role in cell-extracellular matrix interactions.

  • Sequence similarities

    Contains 6 EGF-like domains.
    Contains 4 LDL-receptor class B repeats.
    Contains 1 NIDO domain.
    Contains 1 nidogen G2 beta-barrel domain.
    Contains 1 thyroglobulin type-1 domain.

  • Post-translational
    modifications

    N- and O-glycosylated.

  • Cellular localization

    Secreted > extracellular space > extracellular matrix > basement membrane.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • Enactin antibody
    • Ent antibody
    • Entactin 1 antibody
    • Entactin antibody
    • NID 1 antibody
    • NID antibody
    • NID-1 antibody
    • Nid1 antibody
    • NID1_HUMAN antibody
    • Nidogen (enactin) antibody
    • Nidogen 1 antibody
    • Nidogen antibody
    • Nidogen-1 antibody
    see all

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Entactin/NID antibody [EPR22414-125] (ab254325)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Entactin/NID antibody [EPR22414-125] (ab254325)

    Immunohistochemical analysis of paraffin-embedded rat cardiac muscle tissue labeling Entactin/NID with ab254325 at 1/1000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on basement membrane of rat cardiac muscle (PMID: 10958695). The section was incubated with ab254325 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). 

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Entactin/NID antibody [EPR22414-125] (ab254325)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Entactin/NID antibody [EPR22414-125] (ab254325)

    Immunohistochemical analysis of paraffin-embedded mouse cardiac muscle tissue labeling Entactin/NID with ab254325 at 1/1000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on basement membrane of mouse cardiac muscle (PMID: 10958695). The section was incubated with ab254325 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). 

     

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Frozen sections) - Anti-Entactin/NID antibody [EPR22414-125] (ab254325)
    Immunohistochemistry (Frozen sections) - Anti-Entactin/NID antibody [EPR22414-125] (ab254325)

    Immunohistochemical analysis of 4% paraformaldehyde fixed, 0.2% Triton X-100 permeabilized frozen mouse heart tissue labeling Entactin/NID with ab254325 at 1/500 dilution (green), followed by an AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution. Positive staining on the basement membrane of cardiac muscle (PMID: 12215539) is observed. The nuclear counterstain is DAPI (Blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is an AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution.

  • Immunohistochemistry (Frozen sections) - Anti-Entactin/NID antibody [EPR22414-125] (ab254325)
    Immunohistochemistry (Frozen sections) - Anti-Entactin/NID antibody [EPR22414-125] (ab254325)

    Immunohistochemical analysis of 4% Paraformaldehyde fixed, 0.2% Triton X-100 permeabilized frozen rat heart tissue labeling Entactin/NID with ab254325 at 1/500 dilution (green), followed by an AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution. Positive staining on the basement membrane of cardiac muscle (PMID: 12215539) is observed. The nuclear counterstain is DAPI (Blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is an AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution.

  • Western blot - Anti-Entactin/NID antibody [EPR22414-125] (ab254325)
    Western blot - Anti-Entactin/NID antibody [EPR22414-125] (ab254325)
    All lanes : Anti-Entactin/NID antibody [EPR22414-125] (ab254325) at 1/1000 dilution

    Lane 1 : Mouse heart tissue lysate
    Lane 2 : Mouse lung tissue lysate
    Lane 3 : Mouse skeletal muscle tissue lysate
    Lane 4 : Mouse kidney tissue lysate
    Lane 5 : Rat lung tissue lysate
    Lane 6 : Rat skeletal muscle tissue 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: 136 kDa


    Exposure time: 3 seconds


    Blocking/diluting buffer and concentration: 5% NFDM/TBST 

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:3084254,12215539, 22952693).

  • Western blot - Anti-Entactin/NID antibody [EPR22414-125] (ab254325)
    Western blot - Anti-Entactin/NID antibody [EPR22414-125] (ab254325)
    All lanes : Anti-Entactin/NID antibody [EPR22414-125] (ab254325) at 1/1000 dilution

    Lane 1 : Human kidney tissue lysate
    Lane 2 : Human skeletal muscle tissue lysate
    Lane 3 : Human placenta tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

    Predicted band size: 136 kDa
    Observed band size: 160, 100 kDa why is the actual band size different from the predicted?


    Exposure time: 8 seconds


    Blocking/diluting buffer and concentration: 5% NFDM/TBST

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:3084254, 12215539, 22952693).

  • Western blot - Anti-Entactin/NID antibody [EPR22414-125] (ab254325)
    Western blot - Anti-Entactin/NID antibody [EPR22414-125] (ab254325)
    Anti-Entactin/NID antibody [EPR22414-125] (ab254325) at 1/1000 dilution + Recombinant human Entactin/NID protein (aa29 -1114) 20 ng

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

    Predicted band size: 136 kDa
    Observed band size: 130 kDa why is the actual band size different from the predicted?



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

    Exposure time: 15 seconds.

  • Anti-Entactin/NID antibody [EPR22414-125] (ab254325)
    Anti-Entactin/NID antibody [EPR22414-125] (ab254325)

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

References (1)

Publishing research using ab254325? Please let us know so that we can cite the reference in this datasheet.

ab254325 has been referenced in 1 publication.

  • Jeong S  et al. Anti-Wrinkle Benefits of Peptides Complex Stimulating Skin Basement Membrane Proteins Expression. Int J Mol Sci 21:N/A (2019). PubMed: 31861912

Customer reviews and Q&As

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Entactin/NID antibody [EPR22414-125]

 Excellent
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (Kidney)
Antigen retrieval step
Heat mediated
Permeabilization
No
Specification
Kidney
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: RT°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 May 21 2021

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