HA722119

SEC24A Recombinant Rabbit Monoclonal Antibody [PSH04-36]

cat.: HA722119

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat, Monkey
Applications:	WB, IF-Cell, IHC-P, FC
Clonality:	Monoclonal
Clone number:	PSH04-36

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 120 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within human SEC24A aa 1-350 / 1,093.
Positive control:	Caco-2 cell lysate, HEK-293 cell lysate, HeLa cell lysate, U-87 MG cell lysate, COS-1 cell lysate, Caco-2, human b-cell lymphoma tissue, human stomach tissue, mouse stomach tissue, rat stomach tissue.
Subcellular location:	Cytoplasmic vesicle, COPII-coated vesicle membrane, Endoplasmic reticulum membrane, Cytoplasm, cytosol.
Recommended Dilutions: WB IF-Cell IHC-P FC	1:2,000 1:100 1:200-1:1,000 1:1,000
Uniprot #:	SwissProt: O95486 Human \| Q3U2P1 Mouse Entrez Gene: 287275 Rat
Alternative names:	protein transport protein Sec24A SEC24 family member A S cerevisiae SEC24 related gene family member A SEC24 related protein A SEC24 S cerevisiae related gene family member A

Images

Fig1: Western blot analysis of SEC24A on different lysates with Rabbit anti-SEC24A antibody (HA722119) at 1/2,000 dilution.

Lane 1: Caco-2 cell lysate
Lane 2: HEK-293 cell lysate
Lane 3: HeLa cell lysate
Lane 4: U-87 MG cell lysate
Lane 5: COS-1 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 120 kDa
Observed band size: 120 kDa

Exposure time: 24 seconds; 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 (HA722119) 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.

Fig2: Immunocytochemistry analysis of Caco-2 cells labeling SEC24A with Rabbit anti-SEC24A antibody (HA722119) 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-SEC24A antibody (HA722119) 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.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

	Fig3: Immunohistochemical analysis of paraffin-embedded human b-cell lymphoma tissue with Rabbit anti-SEC24A antibody (HA722119) 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 (HA722119) 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human stomach tissue with Rabbit anti-SEC24A antibody (HA722119) 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 (HA722119) 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded mouse stomach tissue with Rabbit anti-SEC24A antibody (HA722119) at 1/200 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 (HA722119) at 1/200 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.

Fig6: Immunohistochemical analysis of paraffin-embedded rat stomach tissue with Rabbit anti-SEC24A antibody (HA722119) at 1/200 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 (HA722119) at 1/200 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.

Fig7: Flow cytometric analysis of Caco-2 cells labeling SEC24A.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA722119, 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”.