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<strong>Figure 1 Independent Antibody Validation (IAV) via Protein Expression Profile in Human Cell Lines</strong><br>Loading: 15 μg of lysates per lane.Antibodies: SPT2, 6305 (1 μg/mL), SPT2, 64-238 (1 μg/mL), and beta-actin (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 2 Western Blot Validation in Human Cell Lines</strong><br>Loading: 15 μg of lysates per lane.Antibodies: SPT2, 6305 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 3 Western Blot Validation in Mouse 3T3 Cell Lysate in (A) the absence and (B) the presence of blocking peptide</strong><br>Loading: 15 μg of lysates per lane.Antibodies: SPT2, 6305 (0.5 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 4 Western Blot Validation in Human Brain Tissue Lysate</strong><br>Loading: 15 μg of lysates per lane.Antibodies: SPT2, 6305 (Lane 1: 0.5 μg/mL and Lane 2: 1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 5 Western Blot Validation in Mouse 3T3/NIH Cell Lysate</strong><br>Loading: 15 μg of lysates per lane.Antibodies: SPT2, 6305 (Lane 1: 0.5 μg/mL and Lane 2: 1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 6 Immunocytochemistry Validation of SPT2 in Mouse 3T3 Cells</strong><br>Immunocytochemical analysis of 3T3 cells using anti-SPT2 antibody (6305) at 10 μg/ml. Cells was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 7 Immunofluorescence Validation of SPT2 in Mouse 3T3 Cells</strong><br>Immunofluorescent analysis of 4% paraformaldehyde-fixed 3T3 Cells labeling SPT2 with 6305 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
Figure 1 Independent Antibody Validation (IAV) via Protein Expression Profile in Human Cell Lines
Loading: 15 μg of lysates per lane.Antibodies: SPT2, 6305 (1 μg/mL), SPT2, 64-238 (1 μg/mL), and beta-actin (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
IAV
Trial Size

SPT2 Antibody

CATALOG NUMBER: 6305

Clonality:
Polyclonal
Tested Applications:
ELISA, ICC, IF, WB
Host Species:
Rabbit
Species Reactivity:
Human, Mouse
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Mouse
Homology:
Predicted species reactivity based on immunogen sequence: Rat (100%).
Immunogen:
Anti-SPT2 antibody (6305) was raised against a peptide corresponding to 15 amino acids near the carboxy terminus of human SPT2.
The immunogen is located within the last 50 amino acids of SPT2.
Conjugate:
Unconjugated
Tested Applications:
ELISA, ICC, IF, WB
Application Note:
WB: 0.5-1 μg/mL; ICC: 10 μg/mL; IF: 20 μg/mL.
Antibody validated: Western Blot in human and mouse samples; Immunocytochemistry in mouse samples; Immunofluorescence in mouse samples. All other applications and species not yet tested.
Specificity:
SPT2 antibody is predicted to not cross-react with SPT1.
Positive Control 1:
Cat. No. 1282 - 3T3 (NIH) Cell Lysate
Positive Control 2:
Cat. No. 17-201 - 3T3/BALB Cell Slide
Positive Control 3:
Cat. No. 1204 - K562 Cell Lysate
Positive Control 4:
Cat. No. 1219 - MCF7 Cell Lysate
Positive Control 5:
Cat. No. 1220 - SK-N-SH Cell Lysate
Predicted Molecular Weight:
Predicted: 63kD
Observed: 63 kD
Validation:

Independent Antibody Validation in Cell lines (Figure 1) shows similar SPT2 expression profile in human cell lines detected by two independent anti-SPT2 antibodies that recognize different epitopes, 6305 against C-terminus domain and 64-238 against central domain.  SPT2 proteins are detected in the most tested cell lines at different expression levels by the two independent antibodies. 

Isoforms:
Human SPT2 has one isoform (562aa, 63kD). Mouse SPT2 has one isoform (560aa, 63kD) and Rat SPT2 also has one isoform (560aa, 63kD). 6305 can detect human, mouse and rat.
Purification:
SPT2 Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
SPT2 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
SPT2 antibody can be stored at 4°C for three months and -20°C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.
Ncbi Official Symbol:
SPTLC2
Additional Names:
SPT2 Antibody: LCB2, SPT2, HSN1C, LCB2A, NSAN1C, hLCB2a, KIAA0526, LCB2, Serine palmitoyltransferase 2, Long chain base biosynthesis protein 2, LCB 2
Protein Accession Number:
NP_004854
Protein Gi Number:
4758668
Ncbi Gene Id Number:
9517
User Note:
Optimal dilutions for each application to be determined by the researcher.
Background:
SPT2 Antibody: Serine palmitoyltransferase (SPT), which consists of two different subunits, is the key enzyme in sphingolipid biosynthesis. It converts L-serine and palmitoyl-CoA to 3-oxosphinganine with pyridoxal 5'-phosphate as a cofactor. SPT2 is the long chain base subunit 2 of mammalian serine palmitoyltransferase. SPT2 is catalytically active but needs its related protein SPT1 for its stabilization and anchoring the holoenzyme to the cytosolic face of the endoplasmic reticulum. As in the case with SPT1, mutations in the SPT2 gene can cause hereditary sensory and autonomic neuropathy type I (HSAN-I), resulting from a loss of SPT activity as well as the accumulation of the atypical and neurotoxic sphingoid metabolite 1-deoxy-sphinganine.
Background Reference 1:
Batheja et al. J. Histochem. & Cytochem. 2003; 51:687-96.
Background Reference 2:
Nagiec et al. Gene 1996; 177:237-41
Background Reference 3:
Yasuda et al. J. Biol. Chem. 2003; 278:4176-83.
Background Reference 4:
Rotthier et al. Am. J. Hum. Genet. 2010; 87:513-22.

FOR RESEARCH USE ONLY

For additional information, visit ProSci’s Terms & Conditions Page.

Disclaimer:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.

CATALOG NUMBER:

6305

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

Datasheet
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