Antibodies, Proteins, Kits and Reagents for Life Science

314 937 ₸ Product size

100 µl 314 937 ₸

Order now and get it on Tuesday March 02, 2021

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

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 & Species

Application	Species
IHC-Fr	Mouse Rat
IHC-P	Mouse Rat
WB	Mouse Rat Human

See all applications and species data

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.

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: 40% Glycerol, 0.05% BSA, PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR22414-125
Isotype

IgG
Research areas

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)

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)

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)

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)

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)

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)

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)

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