Anti-Chromogranin A antibody [EPR22537-248]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Chromogranin A antibody [EPR22537-248] (AB254322)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized PC-12 (rat adrenal gland pheochromocytoma cell line) cells labeling Chromogranin A with ab254322 at 1/100 dilution, followed by a Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous and cytoplasmic staining in PC-12 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

Secondary antibody only control : Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Chromogranin A antibody [EPR22537-248] (AB254322)

Intracellular flow cytometric analysis of4% paraformaldehyde-fixed, 90% methanol-permeabilized Neuro-2a (mouse neuroblastoma cell line) cells labeling Chromogranin A with ab254322 at 1/60 (red) compared with Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue).

Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Chromogranin A antibody [EPR22537-248] (AB254322)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (mouse neuroblastoma cell line) cells labeling Chromogranin A with ab254322 at 1/100 dilution, followed by a Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous and cytoplasmic staining in Neuro-2a cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

Secondary antibody only control : Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chromogranin A antibody [EPR22537-248] (AB254322)

Immunohistochemical analysis of paraffin-embedded rat panceas tissue labeling Chromogranin A with ab254322 at 1/4000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat pancreas islet (PMID : 22412953) is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

The section was incubated with ab254322 for 30 mins at RT.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-Chromogranin A antibody [EPR22537-248] (AB254322)

Frozen section of 4% PFA-fixed, 0.2% Triton X-100 permeabilized mouse colon tissue labeling Chromogranin A with ab254322 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077), at 1/1000 dilution (green). Positive staining on enteroendocrine cells of mouse colon (PMID : 27287799). is observed. The nuclear counter stain is DAPI (blue).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077), at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-Chromogranin A antibody [EPR22537-248] (AB254322)

Frozen section of 4% PFA-fixed, 0.2% Triton X-100 permeabilized rat colon tissue labeling Chromogranin A with ab254322 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077), at 1/1000 dilution (green). Positive staining on enteroendocrine cells of mouse colon (PMID : 27287799) is observed. The nuclear counter stain is DAPI (blue).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077), at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chromogranin A antibody [EPR22537-248] (AB254322)

Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue labeling Chromogranin A with ab254322 at 1/4000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse pancreas islet (PMID : 22412953) is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

The section was incubated with ab254322 for 30 mins at RT.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• WB

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Western blot - Anti-Chromogranin A antibody [EPR22537-248] (AB254322)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure times.

Lane 1 : 15 seconds; Lane 2 : 3.25 seconds; Lane 3 : 26 seconds; Lane 4 : 3.25 seconds; Lane 5 : 15 seconds.

The molecular weight observed is consistent with what has been described in the literature (PMID : 14734658, 21052719).

All lanes:

Western blot - Anti-Chromogranin A antibody [EPR22537-248] (ab254322) at 1/1000 dilution

Lane 1:

Neuro-2a (mouse neuroblastoma cell line) whole cell lysate at 20 µg

Lane 2:

PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate at 20 µg

Lane 3:

Mouse serum at 20 µg

Lane 4:

Mouse adrenal gland lysate at 20 µg

Lane 5:

Rat adrenal gland lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 50 kDa

Observed band size: 20 kDa,75 kDa

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