Recombinant Anti-PAX5 antibody [SP34] - ChIP Grade (ab227635)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [SP34] to PAX5 - ChIP Grade
- Suitable for: WB, IP, ChIP, IHC-P, Flow Cyt (Intra)
- Reacts with: Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-PAX5 antibody [SP34] - ChIP Grade
See all PAX5 primary antibodies -
Description
Rabbit monoclonal [SP34] to PAX5 - ChIP Grade -
Host species
Rabbit -
Tested applications
Suitable for: WB, IP, ChIP, IHC-P, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- Flow Cyt (Intra): Ramos cells. IHC-P: Human tonsil and B cell lymphoma and rat spleen tissues. IP: Ramos whole cell lysates. WB: Ramos, Daudi, Jurkat and U-937 cell lysates.
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General notes
This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.
Properties
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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: 7.60
Preservative: 0.1% Sodium azide
Constituents: PBS, 1% BSA -
Concentration information loading...
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Purity
Protein A/G purified -
Clonality
Monoclonal -
Clone number
SP34 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab227635 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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WB |
Use a concentration of 0.172 µg/ml. Predicted molecular weight: 42 kDa.
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IP |
1/20.
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ChIP |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration.
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Flow Cyt (Intra) |
1/20.
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Notes |
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WB
Use a concentration of 0.172 µg/ml. Predicted molecular weight: 42 kDa. |
IP
1/20. |
ChIP
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. |
Flow Cyt (Intra)
1/20. |
Target
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Function
May play an important role in B-cell differentiation as well as neural development and spermatogenesis. Involved in the regulation of the CD19 gene, a B-lymphoid-specific target gene. -
Involvement in disease
A chromosomal aberration involving PAX5 is a cause of acute lymphoblastic leukemia. Translocation t(9;18)(p13;q11.2) with ZNF521. Translocation t(9;3)(p13;p14.1) with FOXP1. Translocation t(9;12)(p13;p13) with ETV6.
Leukemia, acute lymphoblastic, 3 -
Sequence similarities
Contains 1 paired domain. -
Developmental stage
Expressed at early B-cell differentiation, in the developing CNS and in adult testis. -
Post-translational
modificationsO-glycosylated. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 5079 Human
- Omim: 167414 Human
- SwissProt: Q02548 Human
- Unigene: 654464 Human
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Alternative names
- B cell lineage specific activator antibody
- B cell lineage specific activator protein antibody
- B cell specific activator protein antibody
see all
Images
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All lanes : Anti-PAX5 antibody [SP34] - ChIP Grade (ab227635) at 0.172 µg/ml
Lane 1 : Ramos (Human Burkitt's lymphoma B lymphocyte) cell lysate
Lane 2 : Daudi (Human Burkitt's lymphoma lymphoblast) cell lysate
Lane 3 : Jurkat (Human T cell leukemia T lymphocyte) cell lysate
Lane 4 : U-937 (Human histiocytic lymphoma monocyte) cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
Lane 1 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 0.05 µg
Lanes 2-4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 0.05 µg/ml
Predicted band size: 42 kDa
Blocking/Diluting buffer and concentration: 5% NFDM /TBST -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat spleen tissue sections labeling PAX5 with ab227635 at 1/50 dilution. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 6.0, epitope retrieval solution 1) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Nuclear staining on the rat spleen, performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab228136 for 30 mins at room temperature. -
Flow Cytometry analysis of Ramos(Human Burkitt's lymphoma B lymphocyte) cells labeling PAX5 with purified ab227635 at 1:20 dilution (8.6 µg/ml) Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor® 488, ab150081) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) / Black. Unlabeled control - Unlabelled cells / Blue.
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Chromatin was prepared from Ramos cells according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min.
The ChIP was performed with 25 µg of chromatin, 5 µg of ab227635 (red), or 5 µg of rabbit normal IgG ab172730 (gray) and 20 µl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci).
Primers and probes are commercial primers from Paper (PMID: 19806612)
*http://www.abcam.com/resources?keywords=X%20ChIP%20protocol -
ab227635 (purified) at 1/20 dilution immunoprecipitating PAX5 in Ramos whole cell lysate.
Lane 1 (input): Ramos(Human Burkitt's lymphoma B lymphocyte) whole cell lysate 10µg
Lane 2 (+): ab227635 & Ramos whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab227635 in Ramos whole cell lysate
For western blotting, ab227648 (purified at 1/500 dilution (0.344 µg/ml) and veriBlot for IP secondary antibody (HRP) (ab131366) at 1/1000 dilution were used.
Blocking and diluting buffer: 5% NFDM /TBST . -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human tonsil tissue sections labeling PAX5 with ab227635 at 1/50 dilution. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 6.0, epitope retrieval solution 1) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Nuclear staining on the human tonsil, performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab228136 for 30 mins at room temperature. -
Immunohistochemical analysis of Human tonsil tissue labeling PAX5 using ab227635.
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Immunohistochemical analysis of Human B cell lymphoma tissue staining PAX5 using ab227635.
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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SDS download
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Datasheet download
References (1)
ab227635 has been referenced in 1 publication.
- Liu PJ et al. In-depth proteomic analysis of six types of exudative pleural effusions for nonsmall cell lung cancer biomarker discovery. Mol Cell Proteomics 14:917-32 (2015). PubMed: 25638566