HA722727

DISC1 Recombinant Rabbit Monoclonal Antibody [JE35-55]

cat.: HA722727

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P, FC, IP
Clonality:	Monoclonal
Clone number:	JE35-55

Form:	Liquid
Storage condition:	Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
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: 94 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within human DISC1 aa 1-150.
Positive control:	HeLa cell lysate, U-87 MG cell lysate, SH-SY5Y cell lysate, Neuro-2a cell lysate, RAW264.7 cell lysate, C6 cell lysate, PC-12 cell lysate, Neuro-2a, C6, human brain tissue, mouse brain tissue, rat brain tissue.
Subcellular location:	Cytoplasm, cytoskeleton, Mitochondrion, microtubule organizing center, centrosome, Postsynaptic density.
Recommended Dilutions: WB IF-Cell IHC-P FC IP	1:1,000 1:100 1:1,000 1:1,000 1-2μg/sample
Uniprot #:	SwissProt: Q9NRI5 Human \| Q811T9 Mouse \| Q810H6 Rat
Alternative names:	C1orf136 DISC1 DISC1_HUMAN Disrupted in schizophrenia 1 Disrupted in schizophrenia 1 protein KIAA0457 RP4-730B13.1 SCZD9

Images

Fig1: Western blot analysis of DISC1 on different lysates with Rabbit anti-DISC1 antibody (HA722727) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: U-87 MG cell lysate
Lane 3: SH-SY5Y cell lysate
Lane 4: Neuro-2a cell lysate
Lane 5: RAW264.7 cell lysate
Lane 6: C6 cell lysate
Lane 7: PC-12 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 94 kDa
Observed band size: 94 kDa

Exposure time: 4 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 (HA722727) at 1/1,000 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 Neuro-2a cells labeling DISC1 with Rabbit anti-DISC1 antibody (HA722727) 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-DISC1 antibody (HA722727) 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: Immunocytochemistry analysis of C6 cells labeling DISC1 with Rabbit anti-DISC1 antibody (HA722727) 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-DISC1 antibody (HA722727) 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-DISC1 antibody (HA722727) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (HA722727) 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 brain tissue with Rabbit anti-DISC1 antibody (HA722727) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (HA722727) 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.

	Fig6: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-DISC1 antibody (HA722727) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (HA722727) 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.
	Fig7: Flow cytometric analysis of Neuro-2a cells labeling DISC1. Cells were fixed and permeabilized. Then stained with the primary antibody (HA722727, 1/1,000) (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).
	Fig8: DISC1 was immunoprecipitated from 0.2 mg Neuro-2a cell lysate with HA722727 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using HA722727 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: Neuro-2a cell lysate (input) Lane 2: HA722727 IP in Neuro-2a cell lysate Lane 3: Rabbit IgG instead of HA722727 in Neuro-2a cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 44 seconds; ECL: K1801

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.