Mouse PU.1/Spi-1 Antibody

Catalog # Availability Size / Price Qty
MAB7124
MAB7124-SP
Mouse PU.1/Spi-1 Antibody in Western Blot
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Mouse PU.1/Spi-1 Antibody Summary

Species Reactivity
Mouse
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
Source
Monoclonal Rat IgG2A Clone # 823123
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
E. coli-derived recombinant mouse Spi-1
Met1-Lys169
Accession # P17433
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.

Applications

Recommended Concentration
Sample
Western Blot
0.1 µg/mL
See below
Immunocytochemistry
8-25 µg/mL
See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Data Examples

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

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.

Immunocytochemistry PU.1/Spi‑1 in J774A.1 Mouse Cell Line. View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

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.

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Preparation and Storage

Reconstitution
Sterile PBS to a final concentration of 0.5 mg/mL.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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: 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.

Long Name
Hematopoietic transcription factor PU.1
Entrez Gene IDs
6688 (Human); 20375 (Mouse); 366126 (Rat)
Alternate Names
hematopoietic transcription factor PU.1; OF; PU.1; SFPI1; SFPI131 kDa transforming protein; SPI-1 proto-oncogene; SPI1; Spi-1; SPI-A; spleen focus forming virus (SFFV) proviral integration oncogene spi1,31 kDa-transforming protein; transcription factor PU.1

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Secondary Antibodies

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