PAX5 was detected in immersion fixed paraffin-embedded sections of human Tonsil using Rabbit Anti-Human PAX5, Monoclonal Antibody (Catalog # MAB9857) at 5ug/mL at 37 ° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module) and Dewax and HIER Buffer H (pH 9; Epredia Catalog # TA-999-DHBH). Tissue was stained using the Alexa Fluor™ Plus 555 Goat anti-Rabbit IgG Secondary Antibody at 1:100 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR555RB) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the nucleus. Protocol available in COMET™ Panel Builder.

Detection of Human Pax5/BSAP by Western Blot.

Western blot shows lysates of Raji human Burkitt's lymphoma cell line, Ramos human Burkitt's lymphoma cell line, Daudi human Burkitt's lymphoma cell line, and Nalm-6 human Pre-B acute lymphocytic leukemia cell line. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-Human/Mouse Pax5/BSAP Monoclonal Antibody (Catalog # MAB3487) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). A specific band was detected for Pax5/BSAP at approximately 42 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Pax5/BSAP in Human PBMCs by Flow Cytometry.

Human peripheral blood mononuclear cells (PBMCs) were stained with Mouse Anti-Human CD19 APC-conjugated Monoclonal Antibody (FAB4867A) and either (A) Rabbit Anti-Human/Mouse Pax5/BSAP Monoclonal Antibody (Catalog # MAB3487) or (B) Normal Rabbit IgG Control (MAB1050) followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (F0110). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (FC012). View our protocol for Staining Intracellular Molecules.

Pax5/BSAP in Mouse Embryo.

Pax5/BSAP was detected in perfusion fixed frozen sections of mouse embryo (13 d.p.c.) using Rabbit Anti-Human/Mouse Pax5/BSAP Monoclonal Antibody (Catalog # MAB3487) at 3 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; CTS005) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Detection of Human Pax5/BSAP by Simple Western^TM.

Simple Western lane view shows lysates of Raji human Burkitt's lymphoma cell line, Ramos human Burkitt's lymphoma cell line, Daudi human Burkitt's lymphoma cell line, and Nalm‑6 human Pre-B acute lymphocytic leukemia cell line, loaded at 0.2 mg/mL. A specific band was detected for Pax5/BSAP at approximately 56 kDa (as indicated) using 1 µg/mL of Rabbit Anti-Human/Mouse Pax5/BSAP Monoclonal Antibody (Catalog # MAB3487). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Pax5/BSAP in Daudi human Burkitt's lymphoma cell line (Positive) & Jurkat human acute T cell leukemia cell line (Negative).

Pax5/BSAP was detected in immersion fixed Daudi human Burkitt's lymphoma cell line (Positive) & Jurkat human acute T cell leukemia cell line (Negative) using Rabbit Anti-Human/Mouse Pax5/BSAP Monoclonal Antibody (Catalog # MAB3487) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to Nucleus. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

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Applications for Human/Mouse Pax5/BSAP Antibody

Application

Recommended Usage

COMET

Flow Cytometry

0.25 µg/10⁶ cells
Sample: Human peripheral blood mononuclear cells (PBMCs) fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012).

Immunocytochemistry

3-25 µg/mL
Sample: Immersion fixed Daudi human Burkitt's lymphoma cell line (Positive) & Jurkat human acute T cell leukemia cell line (Negative)

Immunohistochemistry

3-25 µg/mL
Sample: Perfusion fixed frozen sections of mouse embryo (13 d.p.c.)

Multiplex Immunofluorescence

5 µg/mL
Sample: Immersion fixed paraffin-embedded sections of Human Tonsil.

Simple Western

1 µg/mL
Sample: Raji human Burkitt's lymphoma cell line, Ramos human Burkitt's lymphoma cell line, Daudi human Burkitt's lymphoma cell line, and Nalm‑6 human Pre-B acute lymphocytic leukemia cell line

Western Blot

0.1 µg/mL
Sample: Raji human Burkitt's lymphoma cell line, Ramos human Burkitt's lymphoma cell line, Daudi human Burkitt's lymphoma cell line, and Nalm‑6 human Pre-B acute lymphocytic leukemia cell line

Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Pax5/BSAP

Pax5, also known as BSAP (B-cell-specific transcription factor) is a 42 kDa protein belonging to the paired box transcription factor family. It is a developmental regulator that is important for B-cell lineage commitment and development. Human Pax5 is a 391 amino acid (aa) protein containing the paired DNA-binding domain at the N‑terminal region. Several alternatively spliced isoforms with altered C‑terminal regions and possessing different transactivation properties have been described. All isoforms share the N‑terminal 235 aa with full-length Pax5. Human Pax5 shares 99%, 97% and 94% aa sequence identity with mouse, bovine and canine Pax5, respectively.

Long Name

Paired Box Gene 5/B-cell Specific Activator Protein

Alternate Names

BSAP

Entrez Gene IDs

5079 (Human); 18507 (Mouse)

Gene Symbol

PAX5

UniProt

Q02548

Additional Pax5/BSAP Products

Product Documents for Human/Mouse Pax5/BSAP Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Human/Mouse Pax5/BSAP Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Human/Mouse Pax5/BSAP Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB3487

Key Product Details

Species Reactivity

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Applications

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Label

Antibody Source

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Product Specifications

Immunogen

Specificity

Clonality

Host

Isotype

Scientific Data Images for Human/Mouse Pax5/BSAP Antibody

Detection of PAX5 in Human Tonsil via seqIF™ staining on COMET™​