Ordering Information
- : 05-1000MG
- : 1 mg
Product Images
Phosphoserine Specificity Determined by Blocking Amino Acids:
Calyculin A/Okadaic acid-treated A431 cell lysate resolved by SDS-PAGE, transferred...
Phosphospecificity Demonstrated Through Phosphatase Treatment:
50 ng of recombinant Akt (expressed by baculovirus in Sf21 cells) was either untre...
){kind=spacer}
 was either untreated (Lanes 1 and 3) or pretreated (Lanes 2 and 4) with lambda phosphatase. The two samples in duplicate, were resolved by SDS-PAGE and transferred to PVDF membrane. The membranes were then subsequently probed with either Anti-Akt, PH Domain, clone SKB1 (Cat. No. 05-591, lanes 1 and 2) at 1 μg/mL or Anti-Phosphoserine, clone 4A4 (Cat. No. 05-1000, lanes 3 and 4) at 2 μg/mL.'))
 or 2 µg/mL in 3% milk/TBST (lane 2). Both lanes used 3% milk/TBST for the secondary incubation. '))
 in 3% BSA. Note: expression of various kinases by baculovirus in Sf21 cells can result in basal phosphorylation levels of various serines on the recombinant protein. It is important to note the increase in phosphorylation in the active counterparts. Some of the kinases were not full-length proteins and other with tags, hence they run at a different molecular weight from what would be expected by an endogenous protein of the same target.'))
 in 3% milk. Note: some of the kinases were not full-length proteins, hence they run at a reduced molecular weight.'))
)
 and DAPI (blue).'))
 4A4 staining pattern/morphology on paraffin embedded human breast cancer. Tissue pre-treated with citrate buffer pH 6.0. Antibody diluted to 1:500, IHC HRP/DAB detection system.'))
 or treated (unshaded) with Calyculin A/Okadaic acid (30 minutes). Analysis was run with mouse IgG1 control (purple).'))
Anti-Phosphoserine, clone 4A4
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| Vrt | WB, IF, FC, IH(P), ELISA | Mouse | Purified | Monoclonal Antibody |
Description:
Anti-Phosphoserine Antibody, clone 4A4 | 05-1000MG
Promotional Text:
Trade Name:
Upstate (Millipore)
Specificity:
Serine-phosphorylated proteins from all species
Molecular Weight:
Dependent upon the molecular weight of the serine phosphorylated protein being detected.
Immunogen:
Phosphoserine coupled to KLH
Modifications:
Phosphorylation
Clone:
4A4
Isotype:
IgG1
Background Information:
The identification of protein phosphorylation as a regulatory mechanism originated from studies by Fischer and Krebs in the mid 1950s that later earned them the 1992 Nobel prize. It is the major mechanism for the regulation of diverse cellular processes including cell division, protein synthesis, transcriptional regulation and neurotransmission. The steady state phosphorylation of any given substrate is governed by the opposing activities of kinases and phosphatases. It is now believed that a third of all eukaryotic cellular proteins are phosphorylated and that the majority of all phosphorylation events occur on serine and threonine residues (greater than 95%).
Species Reactivity:
Vertebrates
Control:
Suggested Positive Control: Cat.# 12-628, Calyculin A/Okadaic-treated A431 cell lysate is recommended as a positive control for western immunoblotting. Aliquot as desired, refreeze immediately, and store at –20ºC. The lysate is stable for 6 months at –20ºC.
Quality Assurance:
Routinely evaluated by immunoblot analysis on lysate from Calyculin A/Okadaic-treated human A431 carcinoma cells.
Purification Method:
Protein G-Sepharose™ chromatography
Presentation:
1 mL of PBS
Storage Conditions:
2 years at -20°C from date of shipment.
For maximum recovery of the product, centrifuge the original vial prior to removing the cap. If the product has accidentally been frozen and thawed, spin it at 13,000 x g for 10 minutes at 2-8°C.
For maximum recovery of the product, centrifuge the original vial prior to removing the cap. If the product has accidentally been frozen and thawed, spin it at 13,000 x g for 10 minutes at 2-8°C.
Key Applications:
- Western Blotting
- Immunofluorescence
- Flow Cytometry
- Immunohistochemistry (Paraffin)
- ELISA
Brand Family:
Upstate
Product Name:
Anti-Phosphoserine, clone 4A4
Antibody Type:
Monoclonal Antibody
Qty/Pk:
1 mg
Format:
Purified
Host:
Mouse