Ordering Information
- : 05-1000
- : 100 µg
- : 1 mg/mL
Product Images
Flow Cytometry: Flow cytometry analysis using anti-phosphoserine, clone 4A4. Cells were either untreated (shaded, green) or treated (unshaded) with Ca...
Immunofluorecence Microscopy:
Confocal immunofluorescence image of insulin-treated 293 cells labeled with Anti-phosphoserine, clone 4A4 (green) a...
 or treated (unshaded) with Calyculin A/Okadaic acid (30 minutes). Analysis was run with mouse IgG1 control (purple).'))
 and DAPI (blue).'))
)
 in 3% milk. Note: some of the kinases were not full-length proteins, hence they run at a reduced molecular weight.'))
 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.'))
 or 2 µg/mL in 3% milk/TBST (lane 2). Both lanes used 3% milk/TBST for the secondary incubation. '))
 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.'))
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 or Insulin untreated or treated 293 cell lysate (lanes 3 & 4, respectively) were resolved by electrophoresis, transferred to PVDF, and probed with anti-phosphoserine,4A4 (0.5 µg/mL). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection system.'))
Anti-Phosphoserine, clone 4A4
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| Vrt | ELISA, FC, IF, IH(P), WB | Mouse | Purified | Monoclonal Antibody |
Description:
Anti-Phosphoserine Antibody, clone 4A4 (mouse IgG1) | 05-1000
Promotional Text:
Trade Name:
Upstate (Millipore)
Specificity:
This antibody recognizes 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 (>95%).
Species Reactivity:
Vertebrates
Species Reactivity Note:
Reacts with vertebrates
Application Notes:
Immunofluorescence
Flow Cytometry
Immunohistochemistry (Paraffin)
ELISA
Flow Cytometry
Immunohistochemistry (Paraffin)
ELISA
Control:
Included Positive Control: Cat.# 12-628, Calyculin A/Okadaic-treated A431 cell lysate is provided as a free 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 Western Blot analysis on lysate from Calyculin A/Okadaic-treated human A431 carcinoma cells.
Western Blot Analysis:
0.5–2 μg/mL of this lot detected serinephosphorylated proteins in a lysate from either insulin or Calyculin A/Okadaic-treated human A431 carcinoma cells.
Western Blot Analysis:
0.5–2 μg/mL of this lot detected serinephosphorylated proteins in a lysate from either insulin or Calyculin A/Okadaic-treated human A431 carcinoma cells.
Purification Method:
Protein G-Sepharose™ chromatography
Presentation:
Purified mouse monoclonal IgG1 in buffer containing PBS with 0.1% sodium azide and 30% glycerol.
Storage Conditions:
Stable for 1 year at -20ºC from date of receipt.
For maximum recovery, centrifuge the original vial prior to cap removal. 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, centrifuge the original vial prior to cap removal. If the product has accidentally been frozen and thawed, spin it at 13,000 x g for 10 minutes at 2-8°C.
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Key Applications:
- ELISA
- Flow Cytometry
- Immunofluorescence
- Immunohistochemistry (Paraffin)
- Western Blotting
Brand Family:
Upstate
Product Name:
Anti-Phosphoserine, clone 4A4
Concentration:
1 mg/mL
Antibody Type:
Monoclonal Antibody
Qty/Pk:
100 µg
Format:
Purified
Host:
Mouse