Recombinant Anti-Niemann Pick C2 antibody [EPR19993] KO Tested (ab207158)

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Western blot - Anti-Niemann Pick C2 antibody [EPR19993] (ab207158)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR19993] to Niemann Pick C2
Suitable for: IHC-P, WB
Knockout validated
Reacts with: Mouse, Rat, Human

Carrier Free

Product name

Anti-Niemann Pick C2 antibody [EPR19993]
See all Niemann Pick C2 primary antibodies
Description

Rabbit monoclonal [EPR19993] to Niemann Pick C2
Host species

Rabbit
Tested applications

Suitable for: IHC-P, WBmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human fetal kidney and spleen lysates; HEK-293T, HepG2, C6, RAW 264,7, PC-12 and NIH/3T3 whole cell lysates; mouse spleen lysate; rat kidney lysate. IHC-P: Human endometrium and endometrium cancer tissues; mouse testis tissue; rat kidney 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: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19993
Isotype

IgG
Research areas

Alternative Versions
- Anti-Niemann Pick C2 antibody [EPR19993] - Low endotoxin, Azide free (ab223550)
- Anti-Niemann Pick C2 antibody [EPR19993] - BSA and Azide free (ab271975)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
KO cell lines
- Human NPC2 (Niemann Pick C2) knockout HEK-293T cell line (ab266749)
KO cell lysates
- Human NPC2 (Niemann Pick C2) knockout HEK-293T cell lysate (ab258079)
Recombinant Protein
- Recombinant Human Niemann Pick C2 protein (His tag) (ab200499)
Related Products
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab207158 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
IHC-P		1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. ab93684 can be used for antigen retrieval buffer.
WB		1/1000. Detects a band of approximately 16-18 kDa (predicted molecular weight: 17 kDa).

Notes
IHC-P 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. ab93684 can be used for antigen retrieval buffer.
WB 1/1000. Detects a band of approximately 16-18 kDa (predicted molecular weight: 17 kDa).

Function

May be involved in the regulation of the lipid composition of sperm membranes during the maturation in the epididymis.
Tissue specificity

Epididymis.
Involvement in disease

Defects in NPC2 are the cause of Niemann-Pick disease type C2 (NPDC2) [MIM:607625]. A lysosomal storage disorder that affects the viscera and the central nervous system. It is due to defective intracellular processing and transport of low-density lipoprotein derived cholesterol. It causes accumulation of cholesterol in lysosomes, with delayed induction of cholesterol homeostatic reactions. Niemann-Pick disease type C2 has a highly variable clinical phenotype. Clinical features include variable hepatosplenomegaly and severe progressive neurological dysfunction such as ataxia, dystonia and dementia. The age of onset can vary from infancy to late adulthood.
Sequence similarities

Belongs to the NPC2 family.
Cellular localization

Secreted.
Target information above from: UniProt accession P61916 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 10577 Human
- Entrez Gene: 67963 Mouse
- Entrez Gene: 286898 Rat
- Omim: 601015 Human
- SwissProt: P61916 Human
- SwissProt: Q9Z0J0 Mouse
- Unigene: 433222 Human
- Unigene: 282556 Mouse
Alternative names
- EDDM1 antibody
- Epididymal protein 1 antibody
- Epididymal secretory protein antibody
- Epididymal secretory protein E1 antibody
- hE1 antibody
- Human epididymis-specific protein 1 antibody
- Niemann-Pick disease type C2 antibody
- Niemann-Pick disease type C2 protein antibody
- NPC intracellular cholesterol transporter 2 antibody
- NPC2 antibody
- NPC2_HUMAN antibody
- Tissue specific secretory protein antibody
see all

Western blot - Anti-Niemann Pick C2 antibody [EPR19993] (ab207158)

All lanes : Anti-Niemann Pick C2 antibody [EPR19993] (ab207158) at 1/1000 dilution

Lane 1 : Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lane 2 : NPC2 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lane 3 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 17 kDa
Observed band size: 16-18 kDa why is the actual band size different from the predicted?

Lanes 1-3: Merged signal (red and green). Green - ab207158 observed at 16-18 kDa. Red - loading control ab8245 observed at 36 kDa.

ab207158 Anti-Niemann Pick C2 antibody [EPR19993] was shown to specifically react with Niemann Pick C2 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266749 (knockout cell lysate ab258079) was used. Wild-type and Niemann Pick C2 knockout samples were subjected to SDS-PAGE. ab207158 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993] (ab207158)

Immunohistochemical analysis of paraffin-embedded human endometrium cancer tissue labeling Niemann Pick C2 with ab207158 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human endometrium cancer 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.
Western blot - Anti-Niemann Pick C2 antibody [EPR19993] (ab207158)

Lanes 1 & 3 : Anti-Niemann Pick C2 antibody [EPR19993] (ab207158) at 1/1000 dilution
Lane 2 : Anti-Niemann Pick C2 antibody [EPR19993] (ab207158) at 1/10000 dilution

Lane 1 : Human fetal kidney lysate
Lane 2 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 3 : Human spleen lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 17 kDa
Observed band size: 16-18 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1/2: 3 seconds; Lane 3: 30 seconds.

NPC2 can be glycosylated, the WB pattern is consistent with references PMID:16374838 and PMID:22183894.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993] (ab207158)

Immunohistochemical analysis of paraffin-embedded human endometrium tissue labeling Niemann Pick C2 with ab207158 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human endometrium 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.
Western blot - Anti-Niemann Pick C2 antibody [EPR19993] (ab207158)

All lanes : Anti-Niemann Pick C2 antibody [EPR19993] (ab207158) at 1/1000 dilution

Lane 1 : C6 (Rat glial tumor cell line) whole cell lysate
Lane 2 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 4 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lane 5 : Mouse spleen lysate
Lane 6 : Rat kidney lysate

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 17 kDa
Observed band size: 16-18 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1-3/5: 3 minutes; Lane 4/6: 10 seconds.

NPC2 can be glycosylated, the WB pattern is consistent with references PMID:16374838 and PMID:22183894.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993] (ab207158)

Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling Niemann Pick C2 with ab207158 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on mouse testis 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-Niemann Pick C2 antibody [EPR19993] (ab207158)

Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling Niemann Pick C2 with ab207158 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on rat kidney 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.
Anti-Niemann Pick C2 antibody [EPR19993] (ab207158)

Western blot protocols
Immunohistochemistry protocols

Click here to view the general protocols

Datasheet download

Download

Publishing research using ab207158? Please let us know so that we can cite the reference in this datasheet.

ab207158 has not yet been referenced specifically in any publications.

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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

KO cell lines

KO cell lysates

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Involvement in disease

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

Certificate of Compliance

References (0)

Customer reviews and Q&As