Anti-PSD95抗体- Synaptic Marker
参阅全部 PSD95 一抗
兔多克隆抗体to PSD95 - Synaptic Marker
Rabbit
适用于: ICC/IF, WB, IHC-Pmore details
与反应: Mouse, Rat, Human
预测可用于: Zebrafish, Cynomolgus monkey, Common marmoset
Synthetic peptide corresponding to Mouse PSD95 aa 50-150 (internal sequence) conjugated to keyhole limpet haemocyanin (Cysteine residue).
(Peptide available as ab18661)
WB: Human Brain, Mouse & rat brain tissue lysate, SHSY5Y whole cell lysate . ICC/IF: SH-SY5Y cells , Primary hippocampal rat neurons/glia, (obtained from Neuromics, cat. no. PC35101), DIV14. IHC-P: Mouse & rat brain.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Liquid
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
Immunogen affinity purified
多克隆
IgG
Abpromise™承诺保证使用ab18258于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
| 应用 | Ab评论 | 说明 |
|---|---|---|
| ICC/IF | (5) | Use a concentration of 1 - 5 µg/ml. |
| WB | (14) | Use a concentration of 1 - 3 µg/ml. Detects a band of approximately 85 kDa (predicted molecular weight: 80 kDa). Abcam recommends using 3% milk as the blocking agent. |
| IHC-P | (7) | Use at an assay dependent concentration. |
hide
SwissProt: Q6R005 Zebrafish
Unigene: 463928 Human
Unigene: 27256 Mouse
Unigene: 9765 Rat
Unigene: 30412 Zebrafish
Entrez Gene: 1742 Human
Entrez Gene: 13385 Mouse
Entrez Gene: 405796 Zebrafish
Omim: 602887 Human
SwissProt: P78352 Human
SwissProt: Q62108 Mouse
hide
Dlg4 antibody
DLG4_HUMAN antibody
FLJ97752 antibody
FLJ98574 antibody
Human post synaptic density protein 95 antibody
Post synaptic density protein 95 antibody
Postsynaptic density protein 95 antibody
PSD 95 antibody
PSD-95 antibody
PSD95 antibody
SAP 90 antibody
SAP-90 antibody
SAP90 antibody
Synapse associated protein 90 antibody
Synapse-associated protein 90 antibody
Tax interaction protein 15 antibody
Discs large homolog 4 antibody
Disks large homolog 4 antibody
DLG 4 antibody
Western blot - Anti-PSD95 antibody - Synaptic Marker (ab18258)
All lanes : Anti-PSD95 antibody - Synaptic Marker (ab18258) at 1 µg/ml
Lane 1 : Wild-type U-87 MG cell lysate
Lane 2 : DLG4 knockout U-87 MG cell lysate
Lane 3 : Wild-type SH-SY5Y ab275475 cell lysate
Lane 4 : DLG4 knockout SH-SY5Y ab280043 cell lysate
Lane 5 : Human Brain cell lysate
Lane 6 : HL-60 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 80 kDa
Observed band size: 80 kDa
Western blot: Anti-DLG4 antibody (ab18258) staining at 1 ug/ml, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab18258 was shown to bind specifically to DLG4. A band was observed at 80 kDa in wild-type U-87 MG cell lysates with no signal observed at this size in DLG4 knockout cell line. To generate this image, wild-type and DLG4 knockout U-87 MG cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
Western blot - Anti-PSD95 antibody - Synaptic Marker (ab18258)This image is courtesy of an anonymous Abreview
All lanes : Anti-PSD95 antibody - Synaptic Marker (ab18258) at 1/2000 dilution
All lanes : Human differentiated neurons whole cell lysate
Lysates/proteins at 5 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit HRP at 1/2000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 80 kDa
Observed band size: 100 kDawhy is the actual band size different from the predicted?
Exposure time: 1 minute
Western blot - Anti-PSD95 antibody - Synaptic Marker (ab18258)
All lanes : Anti-PSD95 antibody - Synaptic Marker (ab18258) at 1 µg/ml
Lane 1 : Wild-type U-87 MG cell lysate
Lane 2 : DLG4 knockout U-87 MG cell lysate
Lane 3 : Wild-type SH-SY5Y ab275475 cell lysate
Lane 4 : DLG4 knockout SH-SY5Y ab280043 cell lysate
Lane 5 : Human Brain cell lysate
Lane 6 : HL-60 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 80 kDa
Observed band size: 80 kDa
Western blot: Anti-DLG4 antibody (ab18258) staining at 1 ug/ml, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab18258 was shown to bind specifically to DLG4. A band was observed at 80 kDa in wild-type U-87 MG cell lysates with no signal observed at this size in DLG4 knockout cell line. To generate this image, wild-type and DLG4 knockout U-87 MG cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PSD95 antibody - Synaptic Marker (ab18258)
IHC image of PSD95 staining in a section of formalin-fixed paraffin-embedded normal rat eye performed on a Leica BOND™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab18258, 1ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Immunocytochemistry/ Immunofluorescence - Anti-PSD95 antibody - Synaptic Marker (ab18258)
ab18258 staining PSD95 in primary hippocampal rat neurons/glia, (obtained from Neuromics, cat. no. PC35101), DIV14. cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab18258 at 5µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 4% paraformaldehyde (10 min).
Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.
Western blot - Anti-PSD95 antibody - Synaptic Marker (ab18258)
All lanes : Anti-PSD95 antibody - Synaptic Marker (ab18258) at 1 µg/ml
Lane 1 : Mouse brain tissue lysate at 10 µg
Lane 2 : Human brain tissue lysate at 20 µg
Lane 3 : SHSY5Y whole cell lysate at 20 µg
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution
Predicted band size: 80 kDa
Observed band size: 85 kDawhy is the actual band size different from the predicted?
Exposure time: 8 minutes
Gel type: MOPS
Blocking buffer: 1% milk
Western blot - Anti-PSD95 antibody - Synaptic Marker (ab18258)
All lanes : Anti-PSD95 antibody - Synaptic Marker (ab18258) at 1 µg/ml
Lane 1 : Brain (Mouse) Tissue Lysate (ab27253)
Lane 2 : Brain (Rat) Tissue Lysate (ab7942)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Rabbit IgG secondary antibody (ab28446) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 80 kDa
Observed band size: 85 kDawhy is the actual band size different from the predicted?
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PSD95 antibody - Synaptic Marker (ab18258)This image is courtesy of an abreview submitted by Carl Hobbs, King's College London, United Kingdom
ab18258 (1/500) staining PSD95 in paraffin-embedded mouse cerebellum (top) and medulla (bottom) tissue, showing positive staining to the synaptic regions of the brain. Tissue was fixed in formaldehyde, blocking performed using 1% BSA (10 minutes/RT) and heat mediated antigen retrieval performed before staining. The secondary antibody (1/200) was goat anti rabbit IgG conjugated to Biotin. For further experimental details, please refer to abreview.
Immunocytochemistry/ Immunofluorescence - Anti-PSD95 antibody - Synaptic Marker (ab18258)
ab18258 staining PSD95 in SH-SY5Y cells. The cells were fixed with 100% methanol (5 min) at room temperature, permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with the antibody ab18258 at 5µg/ml and ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PSD95 antibody - Synaptic Marker (ab18258)This image is courtesy of an abreview submitted by Carl Hobbs, King's College London, United Kingdom.
ab18258 (1/1000) staining PSD9 in paraffin-embedded rat cerebellum. Tissue was fixed in formaldehyde, blocking performed using 1% BSA (10 minutes/RT) and heat mediated antigen retrieval performed before staining. The secondary antibody (1/200) was goat anti rabbit IgG conjugated to Biotin. For further experimental details, please refer to abreview.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PSD95 antibody - Synaptic Marker (ab18258)
IHC image of PSD95 staining in mouse brain formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 minutes. The section was then incubated with ab18258, 1 µg/ml, for 15 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Immunocytochemistry/ Immunofluorescence - Anti-PSD95 antibody - Synaptic Marker (ab18258)Image courtesy of an anonymous Abreview.
ab18258 staining PSD95 in human SH-SY5Y cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed in paraformaldehyde, blocked with 10% serum for 20 minutes at 24°C, then incubated with ab18258 at a 1/1000 dilution for 16 hours at 4°C. The secondary used was an Alexa-Fluor® 488 conjugated donkey anti-rabbit polyclonal used at a 1/1000 dilution. Counterstained with Hoechst 33258 (blue).
