SEC24D Recombinant Rabbit Monoclonal Antibody [JE65-27]
Catalog# HA721902
SEC24D Recombinant Rabbit Monoclonal Antibody [JE65-27]
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WB
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IF-Cell
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FC
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IHC-P
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
SEC24D Recombinant Rabbit Monoclonal Antibody [JE65-27]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human SEC24D aa 1-50 / 1032.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, FC, IHC-P
Target Molecular Weight
Predicted band size: 113 kDa
Positive Control
HeLa cell lysate, SH-SY5Y cell lysate, SK-OV-3 cell lysate, RAW264.7 cell lysate, PC-12 cell lysate, HeLa, PC-12, mouse pancreas tissue, rat pancreas tissue.
Conjugation
unconjugated
Clone Number
JE65-27
RRID
Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:2,000
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IF-Cell
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1:100
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FC
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1:1,000
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IHC-P
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1:1,000-1:2,000
Target
Function
Component of the coat protein complex II (COPII) which promotes the formation of transport vesicles from the endoplasmic reticulum (ER). The coat has two main functions, the physical deformation of the endoplasmic reticulum membrane into vesicles and the selection of cargo molecules for their transport to the Golgi complex. Plays a central role in cargo selection within the COPII complex and together with SEC24C may have a different specificity compared to SEC24A and SEC24B. May more specifically package GPI-anchored proteins through the cargo receptor TMED10. May also be specific for IxM motif-containing cargos like the SNAREs GOSR2 and STX5.
Background References
1. Mancias J.D., Goldberg J. The transport signal on Sec22 for packaging into COPII-coated vesicles is a conformational epitope. Mol. Cell 26:403-414 (2007).
2. Bonnon C., Wendeler M.W., Paccaud J.P., Hauri H.P. Selective export of human GPI-anchored proteins from the endoplasmic reticulum. J. Cell Sci. 123:1705-1715 (2010).
3. Mancias J.D., Goldberg J. Structural basis of cargo membrane protein discrimination by the human COPII coat machinery. EMBO J. 27:2918-2928 (2008).
Subcellular Location
Cytoplasm.
Synonyms
KIAA0755 antibody
Protein transport protein Sec24D antibody
SC24D_HUMAN antibody
SEC24 family member D antibody
SEC24 related gene family, member D antibody
SEC24-related protein D antibody
SEC24D antibody
Images
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Western blot analysis of SEC24D on different lysates with Rabbit anti-SEC24D antibody (HA721902) at 1/2,000 dilution.
Lane 1: HeLa cell lysate
Lane 2: SH-SY5Y cell lysate
Lane 3: SK-OV-3 cell lysate
Lane 4: RAW264.7 cell lysate
Lane 5: PC-12 cell lysate
Lysates/proteins at 20 µg/Lane.
Exposure time: 60 seconds; ECL: K1802
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA721902, 1/2,000 in 5% NFDM/TBST, overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 113 kDa
Observed band size: 113 kDa -
☑ Knockdown (KD)
Western blot analysis of SEC24D on different lysates with Rabbit anti-SEC24D antibody (HA721902) at 1/2,000 dilution.
Lane 1: A549-si NT cell lysate
Lane 2: A549-si SEC24D cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 113 kDa
Observed band size: 113 kDa
Exposure time: 3 minutes; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721902) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of HeLa cells labeling SEC24D with Rabbit anti-SEC24D antibody (HA721902) at 1/100 dilution.
Cells were fixed in 100% precooled methanol for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-SEC24D antibody (HA721902) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. -
Immunocytochemistry analysis of PC-12 cells labeling SEC24D with Rabbit anti-SEC24D antibody (HA721902) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-SEC24D antibody (HA721902) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. -
Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue with Rabbit anti-SEC24D antibody (HA721902) at 1/2,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721902) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat pancreas tissue with Rabbit anti-SEC24D antibody (HA721902) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721902) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Flow cytometric analysis of HeLa cells labeling SEC24D.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA721902, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
Flow cytometric analysis of PC-12 cells labeling SEC24D.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA721902, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"