SEC24D Recombinant Rabbit Monoclonal Antibody [JE65-27]
cat.: HA721902
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, FC, IHC-P
Clonality: Monoclonal
Clone number: JE65-27
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 113 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human SEC24D aa 1-50 / 1032.
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.
Subcellular location: Cytoplasm.
Recommended Dilutions:
  WB
  IF-Cell
  FC
  IHC-P
1:2,000
1:100
1:1,000
1:1,000-1:2,000
Uniprot #: SwissProt: O94855 Human
Entrez Gene: 69608 Mouse | 310843 Rat
Alternative names: KIAA0755 Protein transport protein Sec24D SC24D_HUMAN SEC24 family member D SEC24 related gene family, member D SEC24-related protein D SEC24D
Images
HA721902_1.jpg Fig1: 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.

Predicted band size: 113 kDa
Observed band size: 113 kDa

Exposure time: 1 minute; ECL: K1802;

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.
HA721902_2.jpg Fig2: 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.
HA721902_3.jpg Fig3: 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.
HA721902_4.jpg Fig4: 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.
HA721902_5.jpg Fig5: 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.
HA721902_6.jpg Fig6: 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.
HA721902_7.jpg Fig7: 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).
HA721902_8.jpg Fig8: 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).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.