HA724054

CHRNA7 Recombinant Rabbit Monoclonal Antibody [PSH19-20]

cat.: HA724054

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, IHC-Fr, IF-Cell, FC
Clonality:	Monoclonal
Clone number:	PSH19-20

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:	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: 56 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within human CHRNA7 aa 290-502.
Positive control:	Jurkat cell lysate, HeLa cell lysate, A431 cell lysate, Caco-2 cell lysate, RAW264.7 cell lysate, C6 cell lysate, PC-12 cell lysate, Mouse brain tissue lysate, Mouse spleen tissue lysate, Rat brain tissue lysate, Rat spleen tissue lysate, mouse dorsal root ganglion tissue, rat dorsal root ganglion tissue, Jurkat, PC-12.
Subcellular location:	Postsynaptic cell membrane, cell membrane
Recommended Dilutions: WB IHC-P IHC-Fr IF-Cell FC	1:5,000 1:1,000 1:500 1:500 1:1,000
Uniprot #:	SwissProt: P36544 Human \| P49582 Mouse \| Q05941 Rat
Alternative names:	7nAChR a7 nicotinic acetylcholine receptor acetylcholine receptor neuronal nicotinic alpha-7 subunit ACHA7_HUMAN AChR Acra7 alpha 7 neuronal nicotinic acetylcholine receptor alpha-7 nicotinic cholinergic receptor subunit alpha7 cholinergic receptor neuronal nicotinic alpha polypeptide 7 cholinergic receptor nicotinic alpha 7 (neuronal) cholinergic receptor nicotinic alpha 7 cholinergic receptor nicotinic alpha polypeptide 7 CHRNA7 CHRNA7-2 NACHRA7 neuronal acetylcholine receptor protein alpha-7 chain Neuronal acetylcholine receptor subunit alpha-7

Images

Fig1: Western blot analysis of CHRNA7 on different lysates with Rabbit anti-CHRNA7 antibody (HA724054) at 1/5,000 dilution.

Lane 1: Jurkat cell lysate (no heat) (20 µg/Lane)
Lane 2: HeLa cell lysate (no heat) (20 µg/Lane)
Lane 3: A431 cell lysate (no heat) (20 µg/Lane)
Lane 4: Caco-2 cell lysate (no heat) (20 µg/Lane)
Lane 5: RAW264.7 cell lysate (no heat) (20 µg/Lane)
Lane 6: C6 cell lysate (no heat) (20 µg/Lane)
Lane 7: PC-12 cell lysate (no heat) (20 µg/Lane)

Notice: no heat means the lysate is not boiled.

Predicted band size: 56 kDa
Observed band size: 60/48 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 (HA724054) at 1/5,000 dilution was used in primary antibody dilution (K1803) 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: Western blot analysis of CHRNA7 on different lysates with Rabbit anti-CHRNA7 antibody (HA724054) at 1/5,000 dilution.

Lane 1: Jurkat cell lysate (no heat) (20 µg/Lane)
Lane 2: C6 cell lysate (no heat) (20 µg/Lane)
Lane 3: Mouse brain tissue lysate (no heat) (20 µg/Lane)
Lane 4: Mouse spleen tissue lysate (no heat) (20 µg/Lane)
Lane 5: Rat brain tissue lysate (no heat) (20 µg/Lane)
Lane 6: Rat spleen tissue lysate (no heat) (20 µg/Lane)

Notice: no heat means the lysate is not boiled.

Predicted band size: 56 kDa
Observed band size: 60/48 kDa
Exposure time: 59 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 (HA724054) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

	Fig3: Immunohistochemical analysis of paraffin-embedded mouse dorsal root ganglion tissue with Rabbit anti-CHRNA7 antibody (HA724054) 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 (HA724054) 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 rat dorsal root ganglion tissue with Rabbit anti-CHRNA7 antibody (HA724054) 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 (HA724054) 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: Application: IHC-Fr Species: Rat Site: Kidney Sample: Frozen section Antibody concentration: 1/500 Antigen retrieval: Not required

	Fig6: Immunocytochemistry analysis of Jurkat cells labeling CHRNA7 with Rabbit anti-CHRNA7 antibody (HA724054) at 1/500 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-CHRNA7 antibody (HA724054) at 1/500 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 (HA601187, 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.
	Fig7: Immunocytochemistry analysis of PC-12 cells labeling CHRNA7 with Rabbit anti-CHRNA7 antibody (HA724054) at 1/500 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-CHRNA7 antibody (HA724054) at 1/500 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 (HA601187, 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.
	Fig8: Flow cytometric analysis of Jurkat cells labeling CHRNA7. Cells were fixed and permeabilized. Then stained with the primary antibody (HA724054, 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).

Fig9: Flow cytometric analysis of PC-12 cells labeling CHRNA7.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA724054, 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).

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