Neurodegenerative disease-associated synaptic plasticity Antibody Sampler Kit
cat.: HAK21203
Product Type: Antibody Sampler Kit
Species reactivity: Human, Mouse
Applications: WB, IHC-Fr
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
Uniprot #: SwissProt: P17600 Human | Q92777 Human | O88935 Mouse | Q64332 Mouse | P09951 Rat | Q63537 Rat | P14416 Human | P61168 Mouse | P21728 Human | Q61616 Mouse | P18901 Rat | Q13224 Human | Q01097 Mouse | Q00960 Rat | Q05586 Human | P35438 Mouse | P35439 Rat | P42858 Human | P42859 Mouse | P51111 Rat
Images
HAK21203_1.jpg Fig1: Application: Immunofluorescence (IHC-Fr)

Species: Mouse
Tissue: Cerebellum
Sample: Frozen section

Antigen retrieval: Not required

Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 0.5 % Triton X-100 + 0.3 M Glycine in PBS, 10 minutes at room temperature.
Primary antibody: HA723196, 1/500, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.
HAK21203_2.jpg Fig2: Application: Immunofluorescence (IHC-Fr)

Species: Rat
Tissue: Cerebral cortex
Sample: Frozen section

Antigen retrieval: Not required

Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 0.5 % Triton X-100 + 0.3 M Glycine in PBS, 10 minutes at room temperature.
Primary antibody: HA723196, 1/500, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.
HAK21203_3.jpg Fig3: Western blot analysis of Synapsin I + II on different lysates with Rabbit anti-Synapsin I + II antibody (HA723196) at 1/5,000 dilution.

Lane 1: Mouse brain tissue lysate
Lane 2: Mouse lung tissue lysate (negative)
Lane 3: Rat brain tissue lysate
Lane 4: Rat lung tissue lysate (negative)

Lysates/proteins at 20 µg/Lane.

Predicted band size: 74 kDa
Observed band size: 50-74 kDa

Exposure time: 6 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 (HA723196) at 1/5,000 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.
HAK21203_4.jpg Fig4: Immunocytochemistry analysis of mouse primary neuronal cells labeling Synapsin I + II with Rabbit anti-Synapsin I + II antibody (HA723196) 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-Synapsin I + II antibody (HA723196) 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.
HAK21203_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse retina tissue with Rabbit anti-Synapsin I + II antibody (HA723196) 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 (HA723196) 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.
HAK21203_6.jpg Fig6: Immunofluorescence analysis of frozen mouse striatum tissue with Rabbit anti-Dopamine D2 Receptor antibody (HA723162) at 1/500 dilution.

The section was not undergone antigen retrieval. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA723162, green) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
HAK21203_7.jpg Fig7: Western blot analysis of Dopamine D2 Receptor on different lysates with Rabbit anti-Dopamine D2 Receptor antibody (HA723162) at 1/10,000 dilution.

Lane 1: SH-SY5Y cell lysate (no heat)
Lane 2: U-87 MG cell lysate (no heat)
Lane 3: NCI-H1299 cell lysate (no heat)
Lane 4: THP-1 cell lysate (no heat)
Lane 5: Neuro-2a cell lysate (no heat)

Notice: no heat means the lysate is not boiled.

Lysates/proteins at 20 µg/Lane.

Predicted band size: 51 kDa
Observed band size: 60 kDa

Exposure time: 10 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 (HA723162) at 1/10,000 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.
HAK21203_8.jpg Fig8: Immunocytochemistry analysis of SH-SY5Y cells labeling Dopamine D2 Receptor with Rabbit anti-Dopamine D2 Receptor antibody (HA723162) at 1/100 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-Dopamine D2 Receptor antibody (HA723162) 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 (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.
HAK21203_9.jpg Fig9: Application: IHC-Fr

Species: Mouse

Site: Striatum

Sample: Frozen section

Antibody concentration: 1/200

Antigen retrieval: Not required
HAK21203_10.jpg Fig10: Western blot analysis of NMDAR2B on different lysates with Rabbit anti-NMDAR2B antibody (HA722284) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution.

Lane 1: Mouse brain tissue lysate (no heat)
Lane 2: Mouse hippocampus tissue lysate
Lane 3: Mouse liver tissue lysate (negative)
Lane 4: Rat brain tissue lysate (no heat)
Lane 5: Rat hippocampus tissue lysate
Lane 6: Rat liver tissue lysate (negative)
Lane 7: Human brain tissue lysate
Lane 8: Human liver tissue lysate (negative)

Notice: no heat means the lysate is not boiled.

Lysates/proteins at 20 µg/Lane.

Predicted band size: 166 kDa
Observed band size: 115/180 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 (HA722284) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution were 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.
HAK21203_11.jpg Fig11: Immunofluorescence analysis of frozen mouse cerebellum tissue with Rabbit anti-NMDAR2B antibody (HA722284) at 1/500 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA722284, green) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
HAK21203_12.jpg Fig12: Application: IHC-Fr

Species: Mouse

Site: Hippocampus

Sample: Frozen section

Antibody concentration: 1:500

Antigen retrieval: Not required
HAK21203_13.jpg Fig13: Application: IHC-Fr

Species: Mouse

Site: Cerebral cortex

Sample: Frozen section

Antibody concentration: 1:500

Antigen retrieval: Not required
HAK21203_14.jpg Fig14: Application: IHC-Fr

Species: Rat

Site: Cerebral cortex

Sample: Frozen section

Antibody concentration: 1:500

Antigen retrieval: Not required
HAK21203_15.jpg Fig15: Western blot analysis of NMDAR1 on different lysates with Rabbit anti-NMDAR1 antibody (ET1703-75) at 1/5,000 dilution.

Lane 1: MCF7 cell lysate (15 µg/Lane)
Lane 2: Human brain tissue lysate (20 µg/Lane)
Lane 3: Mouse brain tissue lysate (20 µg/Lane)
Lane 4: Rat brain tissue lysate (20 µg/Lane)
Lane 5: Mouse heart tissue lysate (negative) (20 µg/Lane)
Lane 6: Rat liver tissue lysate (negative) (20 µg/Lane)

Predicted band size: 105 kDa
Observed band size: 120 kDa

Exposure time: 1 minute 2 seconds; 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 (ET1703-75) at 1/5,000 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.
HAK21203_16.jpg Fig16: Application: IHC-Fr

Species: Mouse

Site: Cerebral cortex

Sample: Frozen section

Antibody concentration: 1/200

Antigen retrieval: Not required
HAK21203_17.jpg Fig17: Application: IF-tissue

Species: Mouse

Site: Cerebral cortex

Sample: Paraffin-embedded section

Antibody concentration: 1/200
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.