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product-image-AAA126110_FACS8.png FCM/FACS (Flow Cytometry) (Figure 5. Flow Cytometry analysis of THP-1 cells using anti-PU.1/SPI1 antibody (AAA126110).Overlay histogram showing THP-1 cells stained with AAA126110 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PU.1/SPI1 Antibody (AAA126110, 1 ug/1x10^6 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1 ug/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

Rabbit PU.1/SPI1 Polyclonal Antibody | anti-SPI1 antibody

Anti-PU.1/SPI1 Antibody Picoband

Gene Names
SPI1; OF; PU.1; SFPI1; SPI-1; SPI-A
Reactivity
Human, Mouse, Rat
Applications
ELISA, Flow Cytometry, Functional Assay, Immunohistochemistry, Western Blot
Purity
Immunogen affinity purified.
Synonyms
PU.1/SPI1, Antibody; Anti-PU.1/SPI1 Antibody Picoband; anti-SPI1 antibody
Ordering
Host
Rabbit
Reactivity
Human, Mouse, Rat
Clonality
Polyclonal
Isotype
Rabbit IgG
Specificity
Natural and recombinant Mouse Nt5e
Purity/Purification
Immunogen affinity purified.
Form/Format
Lyophilized
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Concentration
Adding 0.2 ml of distilled water will yield a concentration of 500 ug/ml. (varies by lot)
Applicable Applications for anti-SPI1 antibody
ELISA, FCM/FACS (Flow Cytometry), IHC (Immunohistochemistry), WB (Western Blot)
Samples
Cell culture supernatants, serum and plasma (heparin, EDTA)
Detection Range
0.156 ng/ml - 10 ng/ml
Sensitivity
<20 pg/ml
Intra-assay Precision
Intra-Assay Precision (Precision within an assay): Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-assay Precision
Inter-Assay Precision (Precision across assays): Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Preparation and Storage
Store at -20 degree C for one year from date of receipt. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for six months. Avoid repeated freezing and thawing.

FCM/FACS (Flow Cytometry)

(Figure 5. Flow Cytometry analysis of THP-1 cells using anti-PU.1/SPI1 antibody (AAA126110).Overlay histogram showing THP-1 cells stained with AAA126110 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PU.1/SPI1 Antibody (AAA126110, 1 ug/1x10^6 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1 ug/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

product-image-AAA126110_FACS8.png FCM/FACS (Flow Cytometry) (Figure 5. Flow Cytometry analysis of THP-1 cells using anti-PU.1/SPI1 antibody (AAA126110).Overlay histogram showing THP-1 cells stained with AAA126110 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PU.1/SPI1 Antibody (AAA126110, 1 ug/1x10^6 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1 ug/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

IHC (Immunohistochemistry)

(Figure 4. IHC analysis of PU.1/SPI1 using anti-PU.1/SPI1 antibody (AAA126110).PU.1/SPI1 was detected in a paraffin-embedded section of rat lymph gland tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-PU.1/SPI1 Antibody (AAA126110) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)

product-image-AAA126110_IHC10.jpg IHC (Immunohistochemistry) (Figure 4. IHC analysis of PU.1/SPI1 using anti-PU.1/SPI1 antibody (AAA126110).PU.1/SPI1 was detected in a paraffin-embedded section of rat lymph gland tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-PU.1/SPI1 Antibody (AAA126110) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)

IHC (Immunohistochemisry)

(Figure 3. IHC analysis of PU.1/SPI1 using anti-PU.1/SPI1 antibody (AAA126110).PU.1/SPI1 was detected in a paraffin-embedded section of mouse lymph gland tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-PU.1/SPI1 Antibody (AAA126110) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)

product-image-AAA126110_IHC11.jpg IHC (Immunohistochemisry) (Figure 3. IHC analysis of PU.1/SPI1 using anti-PU.1/SPI1 antibody (AAA126110).PU.1/SPI1 was detected in a paraffin-embedded section of mouse lymph gland tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-PU.1/SPI1 Antibody (AAA126110) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)

IHC (Immunohiostchemistry)

(Figure 2. IHC analysis of PU.1/SPI1 using anti-PU.1/SPI1 antibody (AAA126110).PU.1/SPI1 was detected in a paraffin-embedded section of human lymphoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-PU.1/SPI1 Antibody (AAA126110) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)

product-image-AAA126110_IHC13.jpg IHC (Immunohiostchemistry) (Figure 2. IHC analysis of PU.1/SPI1 using anti-PU.1/SPI1 antibody (AAA126110).PU.1/SPI1 was detected in a paraffin-embedded section of human lymphoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-PU.1/SPI1 Antibody (AAA126110) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)

WB (Western Blot)

(Figure 1. Western blot analysis of PU.1/SPI1 using anti-PU.1/SPI1 antibody (AAA126110).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.Lane 1: human K562 whole cell lysates,Lane 2: human Raji whole cell lysates.After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-PU.1/SPI1 antigen affinity purified polyclonal antibody (#AAA126110) at 0.5 ug/mL overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for PU.1/SPI1 at approximately 40 kDa. The expected band size for PU.1/SPI1 is at 40 kDa.)

product-image-AAA126110_WB15.jpg WB (Western Blot) (Figure 1. Western blot analysis of PU.1/SPI1 using anti-PU.1/SPI1 antibody (AAA126110).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.Lane 1: human K562 whole cell lysates,Lane 2: human Raji whole cell lysates.After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-PU.1/SPI1 antigen affinity purified polyclonal antibody (#AAA126110) at 0.5 ug/mL overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for PU.1/SPI1 at approximately 40 kDa. The expected band size for PU.1/SPI1 is at 40 kDa.)
Related Product Information for anti-SPI1 antibody
Background: 5'-nucleotidase (5'-NT), also known as ecto-5'-nucleotidase or CD73 (Cluster of Differentiation 73), is an enzyme that in humans is encoded by the NT5E gene. Ecto-5-prime-nucleotidase catalyzes the conversion at neutral pH of purine 5-prime mononucleotides to nucleosides, the preferred substrate being AMP. The enzyme consists of a dimer of 2 identical 70-kD subunits bound by a glycosyl phosphatidyl inositol linkage to the external face of the plasma membrane. And the enzyme is used as a marker of lymphocyte differentiation. Consequently, a deficiency of NT5 occurs in a variety of immunodeficiency diseases. Other forms of 5-prime nucleotidase exist in the cytoplasm and lysosomes and can be distinguished from ectoNT5 by their substrate affinities, requirement for divalent magnesium ion, activation by ATP, and inhibition by inorganic phosphate. Rareallelic variants are associated with a syndrome of adult-onset calcification of joints and arteries (CALJA) affecting the iliac, femoral, and tibial arteries reducing circulation in the legs and the joints of the hands and feet causing pain.

Principle of the Assay: The Mouse Nt5e Pre-Coated ELISA (Enzyme-Linked Immunosorbent Assay) kit is a solid-phase immunoassay specially designed to measure Mouse Nt5e with a 96-well strip plate that is pre-coated with antibody specific for Nt5e. The detection antibody is a biotinylated antibody specific for Nt5e. The capture antibody is a monoclonal antibody from rat and the detection antibody is a biotinylated polyclonal antibody from goat. The kit contains recombinant Mouse Nt5e with immunogen: Expression system for standard: CHO; Immunogen sequence: W29-S551. The kit is analytically validated with ready-to-use reagents. To measure Mouse Nt5e, add standards and samples to the wells, then add the biotinylated detection antibody. Wash the wells with PBS or TBS buffer, and add Avidin-Biotin-Peroxidase Complex (ABC-HRP). Wash away the unbounded ABC-HRP with PBS or TBS buffer and add TMB. TMB is an HRP substrate and will be catalyzed to produce a blue color product, which changes into yellow after adding the acidic stop solution. The absorbance of the yellow product at 450nm is linearly proportional to Mouse Nt5e in the sample. Read the absorbance of the yellow product in each well using a plate reader, and benchmark the sample wells' readings against the standard curve to determine the concentration of Mouse Nt5e in the sample.
Product Categories/Family for anti-SPI1 antibody

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
31,083 Da
NCBI Official Full Name
transcription factor PU.1 isoform 1
NCBI Official Synonym Full Names
spleen focus forming virus (SFFV) proviral integration oncogene spi1
NCBI Official Symbol
SPI1
NCBI Official Synonym Symbols
OF; PU.1; SFPI1; SPI-1; SPI-A
NCBI Protein Information
transcription factor PU.1; SPI-1 proto-oncogene; 31 kDa transforming protein; 31 kDa-transforming protein; hematopoietic transcription factor PU.1
UniProt Protein Name
Transcription factor PU.1
UniProt Gene Name
SPI1
UniProt Entry Name
SPI1_HUMAN

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Product Notes

The SPI1 spi1 (Catalog #AAA126110) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-PU.1/SPI1 Antibody Picoband reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's PU.1/SPI1 can be used in a range of immunoassay formats including, but not limited to, ELISA, FCM/FACS (Flow Cytometry), IHC (Immunohistochemistry), WB (Western Blot). Researchers should empirically determine the suitability of the SPI1 spi1 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "PU.1/SPI1, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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