Name: Anti-Collagen XVII antibody [EPR18614]
Brand: RabMAb
SKU: ab184996
Rating: 5 (4 reviews)

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Western blot - 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

Alexa Fluor® 647 Carrier Free

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

Suitable for: WB, IHC-P, ICC/IF, IPmore details
Species reactivity

Reacts with: Mouse, Rat, Human
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

Alternative Versions
- Alexa Fluor® 647 Anti-Collagen XVII antibody [EPR18614] (ab218594)
- Anti-Collagen XVII antibody [EPR18614] - BSA and Azide free (ab271950)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab184996 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)	1/1000. Detects a band of approximately 180, 130 kDa (predicted molecular weight: 150 kDa).
IHC-P		1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF	(1)	1/100.
IP	(1)	1/50.

Notes
WB 1/1000. Detects a band of approximately 180, 130 kDa (predicted molecular weight: 150 kDa).
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/100.
IP 1/50.

Relevance

Unlike most collagens, collagen XVII is a transmembrane protein. Collagen XVII is a structural component of hemidesmosomes, multiprotein complexes at the dermal epidermal basement membrane zone that mediate adhesion of keratinocytes to the underlying membrane. Mutations in the gene coding for collagen XVII are associated with both generalized atrophic benign and junctional epidermolysis bullosa. Two homotrimeric forms of type XVII collagen exist. The full length form is the transmembrane protein. A soluble form, referred to as either ectodomain or LAD 1, is generated by proteolytic processing of the full length form. Two transcript variants, one resulting from alternative splicing in the 3' UTR, have been identified for this gene.
Cellular localization

Cell junction, hemidesmosome. Membrane; Single-pass type II membrane protein. Note=Localized along the plasma membrane of the hemidesmosome. 120 kDa linear IgA disease antigen and 97 kDa linear IgA disease antigen: Secreted, extracellular space, extracellular matrix, basement membrane.
Database links
- Entrez Gene: 1308 Human
- Entrez Gene: 12821 Mouse
- Entrez Gene: 294027 Rat
- Omim: 113811 Human
- SwissProt: Q9UMD9 Human
- SwissProt: Q07563 Mouse
Alternative names
- 180 kDa bullous pemphigoid antigen 2 antibody
- Alpha 1 type XVII collagen antibody
- BA16H23.2 antibody
- BP 180 antibody
- BP180 antibody
- BPA 2 antibody
- BPAG 2 antibody
- BPAG2 antibody
- Bullous pemphigoid antigen 2 antibody
- COL17A1 antibody
- Collagen 17 antibody
- Collagen alpha 1 XVII chain antibody
- Collagen alpha 1(XVII) chain antibody
- Collagen alpha1 XVII chain antibody
- Collagen type XVII alpha 1 antibody
- Collagen XVII alpha 1 polypeptide antibody
- CollagenXVII antibody
- Epidermolysis bullosa junctional localisata variant antibody
- FLJ60881 antibody
- KIAA0204 antibody
- LAD 1 antibody
- LAD1 antibody
see all

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)

Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

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Publishing research using ab184996? Please let us know so that we can cite the reference in this datasheet.

ab184996 has been referenced in 25 publications.

Baradaran-Heravi A et al. Nonselective TRPC channel inhibition and suppression of aminoglycoside-induced premature termination codon readthrough by the small molecule AC1903. J Biol Chem 298:101546 (2022). PubMed: 34999117
Ruiz-Torres S et al. Directed differentiation of human pluripotent stem cells into epidermal stem and progenitor cells. Mol Biol Rep 48:6213-6222 (2021). PubMed: 34350551
Nanba D et al. EGFR-mediated epidermal stem cell motility drives skin regeneration through COL17A1 proteolysis. J Cell Biol 220:N/A (2021). PubMed: 34550317
Reske JJ et al. Co-existing TP53 and ARID1A mutations promote aggressive endometrial tumorigenesis. PLoS Genet 17:e1009986 (2021). PubMed: 34941867
Hiroyasu S et al. Granzyme B inhibition reduces disease severity in autoimmune blistering diseases. Nat Commun 12:302 (2021). PubMed: 33436591

View all Publications for this product

Customer reviews and Q&As

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Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (neonatal skin cryosections)

Permeabilization

Yes - 0.1% triton X

Specification

neonatal skin cryosections

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 21°C

Fixative

Formaldehyde

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.

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Submitted Dec 21 2020

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Related Products

Applications

The Abpromise guarantee

Target

Relevance

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (25)

Customer reviews and Q&As

Immunocytochemistry/ Immunofluorescence abreview for Anti-Collagen XVII antibody [EPR18614]