Recombinant Anti-PRAME antibody [EPR20330] - BSA and Azide free (ab232571)
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 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), WB, ICC/IF, IP, IHC-P
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-PRAME antibody [EPR20330] - BSA and Azide free
See all PRAME primary antibodies -
Description
Rabbit monoclonal [EPR20330] to PRAME - BSA and Azide free -
Host species
Rabbit -
Specificity
PRAME is expressed in malignant cells, including leukaemias, Hodgkin's lymphoma, breast cancer, and primary and metastatic melanomas.
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Tested applications
Suitable for: Flow Cyt (Intra), WB, ICC/IF, IP, IHC-Pmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: MeWo and A-375 whole cell lysates; Human ovary cancer and testis lysates. IHC-P: Human melanoma, breast carcinoma and human testis tissue. ICC/IF: MeWo and A-375 cells. Flow Cyt (intra): MeWo cells, K562 cells. IP: MeWo whole cell lysate.
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General notes
ab232571 is the carrier-free version of ab219650.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20330 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab232571 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 |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 57 kDa (predicted molecular weight: 57 kDa).
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ICC/IF |
Use at an assay dependent concentration.
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IP |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Recommend ab219650 incubation at +4°C overnight. |
Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 57 kDa (predicted molecular weight: 57 kDa). |
ICC/IF
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Recommend ab219650 incubation at +4°C overnight. |
Target
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Function
Functions as a transcriptional repressor, inhibiting the signaling of retinoic acid through the retinoic acid receptors RARA, RARB and RARG. Prevents retinoic acid-induced cell proliferation arrest, differentiation and apoptosis. -
Tissue specificity
Expressed in testis. Detected in samples of kidney, brain and skin. -
Sequence similarities
Belongs to the PRAME family.
Contains 4 LRR (leucine-rich) repeats. -
Cellular localization
Nucleus. Cell membrane. - Information by UniProt
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Database links
- Entrez Gene: 23532 Human
- Omim: 606021 Human
- SwissProt: P78395 Human
- Unigene: 30743 Human
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Alternative names
- 4930534P07Rik antibody
- Cancer/testis antigen 130 antibody
- CT130 antibody
see all
Images
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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219650).
Flow cytometry overlay histogram showing left K562 positive cells and right negative HEK293 stained with ab219650 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1 % PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab219650) (1x 106 in 100μl at 0.008μg/ml (1/267500)) for 30min at 22°C.
The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C
Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
This antibody gave a positive signal in K562 Fixed with 80% methanol (5 min) / permeabilised with 0.1 % PBS-Triton X-100 for 15 min under the same conditions.
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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219650).
Immunohistochemical analysis of paraffin-embedded human melanoma tissue labeling PRAME with ab219650 at 1/500 dilution, followed by LeicaDS9800 (Bond™ Polymer Refine Detection). Sections were counter stained with Hematoxylin. Antigen retrieval was heat mediated with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Nuclear staining on human melanoma. The section was incubated with ab219650 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.Secondary antibody only control: Used PBS instead of primary antibody.
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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219650).
ab219650 staining PRAME in K562 cells, with negative expression in Ramos cells. The cells were fixed with 4% formaldehyde (10 min), permeabilised with 0.1% Triton x-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab219650 at 1 µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin at 0.5 µg/ml. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150119, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 647), pre-adsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.
This product also work with 100% methanol (5 min) fixation under the same testing conditions.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed and 90% Methanol-permeabilised 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219650).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219650).
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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219650).
Immunohistochemical analysis of paraffin-embedded human testis tissue labeling PRAME with ab219650 at 1/500 dilution, followed by LeicaDS9800 (Bond™ Polymer Refine Detection). Sections were counter stained with Hematoxylin. Antigen retrieval was heat mediated with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Nuclear staining on human testis. The section was incubated with ab219650 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.Secondary antibody only control: Used PBS instead of primary antibody.
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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219650).
Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue labeling PRAME with ab219650 at 1/500 dilution, followed by LeicaDS9800 (Bond™ Polymer Refine Detection). Sections were counter stained with Hematoxylin. Antigen retrieval was heat mediated with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Nuclear staining on human breast carcinoma. The section was incubated with ab219650 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.Secondary antibody only control: Used PBS instead of primary antibody.
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219650).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219650).
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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219650).
Immunohistochemical analysis of paraffin-embedded human stomach tissue labeling PRAME with ab219650 at 1/500 dilution, followed by LeicaDS9800 (Bond™ Polymer Refine Detection). Sections were counter stained with Hematoxylin. Antigen retrieval was heat mediated with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Negative control: No staining on human stomach. The section was incubated with ab219650 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219650).
Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling PRAME with ab219650 at 1/500 dilution, followed by LeicaDS9800 (Bond™ Polymer Refine Detection). Sections were counter stained with Hematoxylin. Antigen retrieval was heat mediated with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Negative control: No staining on human tonsil. The section was incubated with ab219650 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219650).
Immunohistochemical analysis of paraffin-embedded human breast tissue labeling PRAME with ab219650 at 1/500 dilution, followed by LeicaDS9800 (Bond™ Polymer Refine Detection). Sections were counter stained with Hematoxylin. Antigen retrieval was heat mediated with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Negative control: No staining on human breast. The section was incubated with ab219650 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
Tissue Microarrays stained for " Anti-PRAME antibody [EPR20330]” using " ab219650" in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). The sections were incubated with ab219650 at +4°C overnight followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab232571 has not yet been referenced specifically in any publications.