Anti-GAP43 抗体 [EP890Y] - BSA and Azide free

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-GAP43 antibody [EP890Y] - BSA and Azide free (AB219582)

ICC/IF image of ab75810 (unpurified) stained SKNSH cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab75810, 1/50 dilution) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG(H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75810).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-GAP43 antibody [EP890Y] - BSA and Azide free (AB219582)

Clone EP890Y (ab219582) has been successfully conjugated by Abcam. This image was generated using Anti-GAP43 antibody [EP890Y] (Alexa Fluor® 488). Please refer to ab196324 for protocol details.

ab196324 staining GAP43 in U87MG cells. The cells were fixed with 100% methanol (5 min), 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 overnight at +4°C with ab196324 at a 1/100 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 594), at a 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

This product also gave a positive signal under the same testing conditions in U87MG cells fixed with 4% formaldehyde (10 min).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAP43 antibody [EP890Y] - BSA and Azide free (AB219582)

This data was developed using ab75810 the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of formalin fixed paraffin embedded human cerebrum labelling GAP43 with ab75810 at a concentration of 0.1µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32 mins. ab75810 anti-GAP43 antibody [EP890Y] was incubated for 16 mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-GAP43 antibody [EP890Y] - BSA and Azide free (AB219582)

Clone EP890Y (ab219582) has been successfully conjugated by Abcam. This image was generated using Anti-GAP43 antibody [EP890Y] (Alexa Fluor® 647). Please refer to ab196540 for protocol details.

ab196540 staining GAP43 in U87MG cells. The cells were fixed with 100% methanol (5 min), 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 overnight at +4°C with ab196540 at a 1/100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at a 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

This product also gave a positive signal under the same testing conditions in U87MG cells fixed with 4% formaldehyde (10 min).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-GAP43 antibody [EP890Y] - BSA and Azide free (AB219582)

Overlay histogram showing SH-SY5Y cells stained with ab75810 (unpurified) (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab75810, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75810).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-GAP43 antibody [EP890Y] - BSA and Azide free (AB219582)

Clone EP890Y (ab219582) has been successfully conjugated by Abcam. This image was generated using Anti-GAP43 antibody [EP890Y] (PE). Please refer to ab208745 for protocol details.

ab208745 staining GAP43 in u87mg cells. The cells were fixed with 100% methanol (5min), 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 overnight at +4°C with ab208745 at 1/100 dilution (Pseudocolored in green) and ab195884, Rat monoclonal to Tubulin (Alexa Fluor^® 647), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-GAP43 antibody [EP890Y] - BSA and Azide free (AB219582)

Intracellular Flow Cytometry analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling GAP43 with Purified ab75810 at 1/20 dilution (10 μg/ml) (Red). Cells were fixed with 80% Methanol and permeabilised with 0.1% Tween-20. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue). This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75810)

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAP43 antibody [EP890Y] - BSA and Azide free (AB219582)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebrum tissue sections labeling GAP43 with Purified ab75810 at 1 : 3000 dilution (0.07 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used for detection. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75810)

• IP

Lab

Immunoprecipitation - Anti-GAP43 antibody [EP890Y] - BSA and Azide free (AB219582)

ab75810 (Purified) at 1 : 20 dilution (1 μg) immunoprecipitating GAP43 in SH-SY5Y whole cell lysate.
Lane 1 (input) : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate 10 μg
Lane 2 (+) : ab75810 & SH-SY5Y whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab75810 in SH-SY5Y whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1 : 1000 dilution.
Blocking and diluting buffer : 5% NFDM/TBST.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75810)

All lanes:

Immunoprecipitation - Anti-GAP43 antibody [EP890Y] - Neuronal Marker (<a href='/products/primary-antibodies/gap43-antibody-ep890y-neuronal-marker-ab75810'>ab75810</a>)

Predicted band size: 24 kDa,45 kDa

false

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAP43 antibody [EP890Y] - BSA and Azide free (AB219582)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat cerebrum tissue sections labeling GAP43 with Purified ab75810 at 1 : 3000 dilution (0.07 μg/ml). Heat mediated antigen retrieval using Bond© Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used for detection. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75810)

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-GAP43 antibody [EP890Y] - BSA and Azide free (AB219582)

Immunocytochemistry/ Immunofluorescence analysis of Neuro-2a (Mouse neuroblastoma neuroblast) cells labeling GAP43 with Purified ab75810 at 1 : 160 dilution (1.4 µg/ml). Cells were fixed in 100% Methanol and permeabilized with None. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1 : 200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1 : 1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75810)

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAP43 antibody [EP890Y] - BSA and Azide free (AB219582)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cerebrum tissue sections labeling GAP43 with Purified ab75810 at 1 : 3000 dilution (0.07 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used for detection. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75810)

• IHC-P

AbReview18768****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAP43 antibody [EP890Y] - BSA and Azide free (AB219582)

ab75810 (unpurified) staining GAP43 in Mouse ear tissue sections by Immunohistochemistry (Formalin/ PFA-fixed paraffin-embedded tissue sections). The sections were formaldehyde fixed, subjected to heat mediated antigen retrieval at pH 6 and blocked for 10 minutes at 25C. The primary antibody was diluted 1/500 and incubated with the sample for 1 hour at 25°C. An HRP polymer anti-rabbit IgG system was used undiluted, as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75810).

This image is courtesy of an anonymous Abreview.