Antibodies, Proteins, Kits and Reagents for Life Science

311 587 ₸ Product size

100 µl 311 587 ₸

Order now and get it on Tuesday March 02, 2021

Anti-PRAME antibody [EPR20330] (ab219650)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR20330] to PRAME
Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
Reacts with: Human

Product name

Anti-PRAME antibody [EPR20330]
See all PRAME primary antibodies
Description

Rabbit monoclonal [EPR20330] to PRAME
Host species

Rabbit
Specificity

PRAME is expressed in malignant cells, including leukaemias, Hodgkin's lymphoma, breast cancer, and primary and metastatic melanomas.

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IHC-P	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: MeWo and A-375 whole cell lysates; Human ovary cancer and testis lysates. IHC-P: Human testis and melanoma tissues. ICC/IF: MeWo and A-375 cells. Flow Cyt: MeWo cells. IP: MeWo whole cell lysate.
General notes
PRAME (PReferentially expressed Antigen in MElanoma) is a tumor-associated antigen and is a member of the family of cancer testis antigens (CTA). PRAME is expressed in malignant cells, including leukaemias, Hodgkin's lymphoma, breast cancer, and primary and metastatic melanomas. For more information, please refer to PMID: 27441500. PRAME has low or no expression in normal tissues except for in testis, ovary, placenta, adrenals and endometrium. For more information, please refer to PMID: 9047241.
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

EPR20330
Isotype

IgG
Research areas

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRAME antibody [EPR20330] (ab219650)

Immunohistochemical analysis of paraffin-embedded human melanoma tissue labeling PRAME with ab219650 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining on human melanoma (PMID: 9047241).

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-PRAME antibody [EPR20330] (ab219650)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MeWo (Human malignant melanoma cell line) cells labeling PRAME with ab219650 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing mostly nuclear staining on MeWo cells.

The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.
Flow Cytometry - Anti-PRAME antibody [EPR20330] (ab219650)

Flow cytometric analysis of 4% paraformaldehyde-fixed MeWo (Human malignant melanoma cell line) cells labeling PRAME with ab219650 at 1/500 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
Western blot - Anti-PRAME antibody [EPR20330] (ab219650)

All lanes : Anti-PRAME antibody [EPR20330] (ab219650) at 1/1000 dilution

Lane 1 : Human ovary cancer lysate
Lane 2 : A-375 (Human malignant melanoma cell line) whole cell lysate
Lane 3 : Human testis 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: 57 kDa
Observed band size: 57 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1: 3 minutes; Lane 2: 5 seconds; Lane 3: 1 minute.
Immunoprecipitation - Anti-PRAME antibody [EPR20330] (ab219650)

PRAME was immunoprecipitated from 0.35 mg of MeWo (Human malignant melanoma cell line) whole cell lysate with ab219650 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab219650 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: MeWo whole cell lysate 10 μg (Input).

Lane 2: ab219650 IP in MeWo whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab219650 in MeWo whole cell lysate.

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

Exposure time: 1 second.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRAME antibody [EPR20330] (ab219650)

Immunohistochemical analysis of paraffin-embedded human testis tissue labeling PRAME with ab219650 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining on human testis (PMID: 9047241).

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-PRAME antibody [EPR20330] (ab219650)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A-375 (Human malignant melanoma cell line) cells labeling PRAME with ab219650 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing mostly nuclear staining on A-375 cells.

The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.
Western blot - Anti-PRAME antibody [EPR20330] (ab219650)

Anti-PRAME antibody [EPR20330] (ab219650) at 1/1000 dilution + MeWo (Human malignant melanoma cell line) whole cell lysate at 20 µg

Secondary
Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/4000 dilution

Predicted band size: 57 kDa
Observed band size: 57 kDa

Exposure time: 15 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRAME antibody [EPR20330] (ab219650)

Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling PRAME with ab219650 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Negative control: No staining on human pancreas (PMID: 9047241).

Counter stained with Hematoxylin.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Anti-PRAME antibody [EPR20330] (ab219650)

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