SPT2 Antibody

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ProSci
Product Code: PSI-6305
Product Group: Primary Antibodies
Supplier: ProSci
CodeSizePrice
PSI-6305-0.02mg0.02mg£150.00
Quantity:
PSI-6305-0.1mg0.1mg£449.00
Quantity:
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Overview

Host Type: Rabbit
Antibody Isotype: IgG
Antibody Clonality: Polyclonal
Regulatory Status: RUO
Applications:
  • Enzyme-Linked Immunosorbent Assay (ELISA)
  • Immunofluorescence (IF)
  • Immunohistochemistry (IHC)
  • Western Blot (WB)

Images

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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.
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<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.
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<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.
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<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.
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<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.
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<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.
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<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).
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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).

Documents

Further Information

Additional Names:
SPT2 Antibody: LCB2, SPT2, HSN1C, LCB2A, NSAN1C, hLCB2a, KIAA0526, LCB2, Serine palmitoyltransferase 2, Long chain base biosynthesis protein 2, LCB 2
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.
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 References:
  • Batheja et al. J. Histochem. & Cytochem. 2003; 51:687-96.
  • Nagiec et al. Gene 1996; 177:237-41
  • Yasuda et al. J. Biol. Chem. 2003; 278:4176-83.
  • Rotthier et al. Am. J. Hum. Genet. 2010; 87:513-22.
Buffer:
SPT2 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Conjugate:
Unconjugated
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.
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.
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.
NCBI Gene ID #:
9517
NCBI Official Name:
serine palmitoyltransferase, long chain base subunit 2
NCBI Official Symbol:
SPTLC2
NCBI Organism:
Homo sapiens
Physical State:
Liquid
PREDICTED MOLECULAR WEIGHT:
Predicted: 63kD

Observed: 63 kD
Protein Accession #:
NP_004854
Protein GI Number:
4758668
Purification:
SPT2 Antibody is affinity chromatography purified via peptide column.
Research Area:
Homeostasis
SPECIFICITY:
SPT2 antibody is predicted to not cross-react with SPT1.
Swissprot #:
O15270
User NOte:
Optimal dilutions for each application to be determined by the researcher.
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. 

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