Antibodies, Proteins, Kits and Reagents for Life Science

321 638 ₸ Product size

100 µl 321 638 ₸

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Anti-Collagen XVII antibody [EPR18614] (ab184996)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR18614] to Collagen XVII
Suitable for: WB, IHC-P, ICC/IF, IP
Reacts with: Mouse, Rat, Human

Product name

Anti-Collagen XVII antibody [EPR18614]
See all Collagen XVII primary antibodies
Description

Rabbit monoclonal [EPR18614] to Collagen XVII
Host species

Rabbit

Tested Applications & Species

Application	Species
ICC/IF	Human
IHC-P	Mouse Rat Human
IP	Human
WB	Mouse Rat Human

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: A431 whole cell lysate, human, mouse and rat skin lysates. IHC-P: Human esophagus, mouse skin and rat skin tissues. ICC/IF: HaCaT and A431 cells. IP: A431 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.

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: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18614
Isotype

IgG
Research areas

Western blot - Anti-Collagen XVII antibody [EPR18614] (ab184996)

All lanes : Anti-Collagen XVII antibody [EPR18614] (ab184996) at 1/5000 dilution

Lane 1 : Human skin lysate
Lane 2 : Human fetal liver lysate
Lane 3 : Human fetal lung lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 150 kDa
Observed band size: 130,180 kDa
why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 15161638).
Western blot - Anti-Collagen XVII antibody [EPR18614] (ab184996)

Anti-Collagen XVII antibody [EPR18614] (ab184996) at 1/5000 dilution + A431 (Human epidermoid carcinoma) whole cell lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 150 kDa
Observed band size: 130,180 kDa why is the actual band size different from the predicted?

Exposure time: 1 minute

Blocking/Dilution buffer: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 15161638).
Western blot - Anti-Collagen XVII antibody [EPR18614] (ab184996)

Anti-Collagen XVII antibody [EPR18614] (ab184996) at 1/2000 dilution + Mouse skin lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 150 kDa
Observed band size: 130,180 kDa why is the actual band size different from the predicted?

Exposure time: 2 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 15161638).
Western blot - Anti-Collagen XVII antibody [EPR18614] (ab184996)

Anti-Collagen XVII antibody [EPR18614] (ab184996) at 1/1000 dilution + Rat skin lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 150 kDa
Observed band size: 130,180 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 15161638).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen XVII antibody [EPR18614] (ab184996)

Immunohistochemical analysis of paraffin-embedded Human esophagus tissue labeling Collagen XVII with ab184996 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on squamous epithelial cells of Human esophagus is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

J Biol Chem. 1991. 266, 24064-24069.

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-Collagen XVII antibody [EPR18614] (ab184996)

Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling Collagen XVII with ab184996 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Negative on Human pancreas. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

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-Collagen XVII antibody [EPR18614] (ab184996)

Immunohistochemical analysis of paraffin-embedded mouse skin tissue labeling Collagen XVII with ab184996 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic and membrane staining on squamous epithelial cells of mouse skin is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

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-Collagen XVII antibody [EPR18614] (ab184996)

Immunohistochemical analysis of paraffin-embedded rat skin tissue labeling Collagen XVII with ab184996 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on epithelial cells of rat skin is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-Collagen XVII antibody [EPR18614] (ab184996)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HaCaT (Human keratinocyte cells) cells labeling Collagen XVII with ab184996 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and membranous staining on HaCaT cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab184996 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2:. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-Collagen XVII antibody [EPR18614] (ab184996)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A431 (Human epidermoid carcinoma) cells labeling Collagen XVII with ab184996 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and membranous staining on A431 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab184996 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.
Immunoprecipitation - Anti-Collagen XVII antibody [EPR18614] (ab184996)

Collagen XVII was immunoprecipitated from 1mg of A431 (Human epidermoid carcinoma) whole cell lysate with ab184996 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab184996 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: A431 whole cell lysate 10ug (Input).

Lane 2: ab184996 IP in A431 whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab184996 in A431 whole cell lysate.

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

Exposure time: 10 seconds.
Anti-Collagen XVII antibody [EPR18614] (ab184996)

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