Mouse PU.1/Spi-1 Antibody (823123)

Name: Mouse PU.1/Spi-1 Antibody (823123)
Brand: R&D Systems
SKU: MAB7124
Price: 449 USD
Availability: InStock

R&D Systems | Catalog # MAB7124

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Product Documents

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Product Specifications
Scientific Data
Applications
Preparation & Storage
Calculators
Background
Product Documents
Specific Notices
Research Areas

Key Product Details

Species Reactivity

Validated:

Mouse

Cited:

Mouse

Applications

Validated:

Western Blot, Immunocytochemistry

Cited:

Immunohistochemistry

Label

Unconjugated

Antibody Source

Monoclonal Rat IgG_2A Clone # 823123

View all PU.1/Spi-1 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant mouse Spi-1
Met1-Lys169
Accession # P17433

Specificity

Detects mouse PU.1/Spi-1 in ELISAs. In direct ELISAs, no cross-reactivity with recombinant human PU.1/Spi-1 is observed. Detects mouse and rat PU.1/Spi-1 in Western Blots

Clonality

Monoclonal

Host

Rat

Isotype

IgG_2A

Scientific Data Images for Mouse PU.1/Spi-1 Antibody (823123)

Detection of Mouse and Rat PU.1/Spi‑1 by Western Blot.

Western blot shows lysates of NR8383 rat alveolar macrophage cell line and J774A.1 mouse reticulum cell sarcoma macrophage cell line. PVDF membrane was probed with 0.1 µg/mL of Rat Anti-Mouse PU.1/Spi-1 Monoclonal Antibody (Catalog # MAB7124) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for PU.1/Spi-1 at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

PU.1/Spi‑1 in J774A.1 Mouse Cell Line.

PU.1/Spi-1 was detected in immersion fixed J774A.1 mouse reticulum cell sarcoma macrophage cell line using Rat Anti-Mouse PU.1/Spi-1 Monoclonal Antibody (Catalog # MAB7124) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red, upper panel; Catalog # NL013) and counterstained with DAPI (blue, lower panel). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Applications for Mouse PU.1/Spi-1 Antibody (823123)

Application

Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed J774A.1 mouse reticulum cell sarcoma macrophage cell line

Western Blot

0.1 µg/mL
Sample: NR8383 rat alveolar macrophage cell line and J774A.1 mouse reticulum cell sarcoma macrophage cell line

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

Volume (to add to vial)

Mass (in vial)

Desired Reconstitution Concentration

Background: PU.1/Spi-1

Spi-1 (SFFV proviral integration 1 protein; also PU.1 and Sfpi1) is a 37-41 kDa member of the ets family of transcription factors. It is both an RNA and DNA binding protein that is found in hematopoietic cells such as B cells, neutrophils, macrophages and dendritic cells (DC). Spi-1 can act as both a transcriptional activator and repressor. In DC, Spi-1 promotes the expression of CD80, CD86 and CD11b, while in proerythroblasts it blocks GATA-1 induced transcriptional activation. Spi-1 binds to DNA as a monomer and interacts with multiple factors such as Runx-1, IRF8, GATA-1 and c-Jun. Mouse Spi-1 is 272 amino acids (aa) in length. It contains a transactivation region (aa 7-117), a PEST domain (aa 118-164), and a DNA-binding domain (aa 165-266). Phosphorylation on Ser41, 142 and 148 increases activity. There is one alternative start site at Met7. Over aa 1-169, mouse Spi-1 shares 88% and 81% aa identity with rat and human Spi-1, respectively.