HA721892

SNAP25 Recombinant Rabbit Monoclonal Antibody [JE45-86]

cat.: HA721892

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, IF-Cell, FC
Clonality:	Monoclonal
Clone number:	JE45-86

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: 23 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within Human SNAP25 aa 51-150 / 206.
Positive control:	SH-SY5Y cell lysate, Neuro-2a cell lysate, human brain tissue lysate, SH-SY5Y, human brain tissue, mouse brain tissue, rat brain tissue.
Subcellular location:	Cytoplasm, perinuclear region. Cell membrane.
Recommended Dilutions: WB IHC-P IF-Cell FC	1:2,000 1:500-1:5,000 1:100 1:1,000
Uniprot #:	SwissProt: P60880 Human \| P60879 Mouse \| P60881 Rat
Alternative names:	bA416N4.2 Bdr CMS18 dJ1068F16.2 FLJ23079 HGNC:11132 MGC105414 MGC139754 Resistance to inhibitors of cholinesterase 4 homolog RIC 4 RIC-4 RIC4 SEC 9 SEC9 SNAP 25 SNAP SNAP-25 SNAP-25B SNAP25 SNP 25 SNP25 SNP25_HUMAN sp SUP Super protein Synaptosomal associated 25 kDa protein Synaptosomal associated protein Synaptosomal associated protein 25 Synaptosomal associated protein 25kDa Synaptosomal-associated 25 kDa protein Synaptosomal-associated protein 25 Synaptosomal-associated protein Synaptosomal-associated protein, 25-KD

Images

Fig1: Western blot analysis of SNAP25 on different lysates with Rabbit anti-SNAP25 antibody (HA721892) at 1/2,000 dilution.

Lane 1: SH-SY5Y cell lysate (20 µg/Lane)
Lane 2: Neuro-2a cell lysate (20 µg/Lane)
Lane 3: K-562 cell lysate (negative) (20 µg/Lane)
Lane 4: Human brain tissue lysate (40 µg/Lane)

Predicted band size: 23 kDa
Observed band size: 25 kDa

Exposure time: 1 minute 21 seconds;

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 (HA721892) 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 SH-SY5Y cells labeling SNAP25 with Rabbit anti-SNAP25 antibody (HA721892) 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-SNAP25 antibody (HA721892) 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 brain tissue with Rabbit anti-SNAP25 antibody (HA721892) at 1/500 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 (HA721892) at 1/500 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 mouse brain tissue with Rabbit anti-SNAP25 antibody (HA721892) 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 (HA721892) 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-SNAP25 antibody (HA721892) at 1/5,000 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 (HA721892) at 1/5,000 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.

Fig6: Immunohistochemical analysis of paraffin-embedded human colon tissue (negative) with Rabbit anti-SNAP25 antibody (HA721892) at 1/500 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 (HA721892) at 1/500 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 SH-SY5Y cells labeling SNAP25.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721892, 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”.