Anti-Chromogranin A antibody [RM1025]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Chromogranin A antibody [RM1025] (AB283265)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Jurkat (Human T cell leukemia T lymphocyte / Left) and SH-SY5Y (Human neuroblastoma epithelial cell / Right) cells labelling Chromogranin A with ab283265 at 1/500 dilution (0.1 μg) compared with an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

Negative control : Jurkat.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chromogranin A antibody [RM1025] (AB283265)

Immunohistochemical analysis of paraffin-embedded human stomach tissue labelling Chromogranin A with ab283265 at 1/5000 dilution (0.107 μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on the endocrine cells in human stomach. The section was incubated with ab283265 overnight at 4℃. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

Heat mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0)

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chromogranin A antibody [RM1025] (AB283265)

Immunohistochemical analysis of paraffin-embedded human pancreas tissue labelling Chromogranin A with ab283265 at 1/5000 dilution (0.107 μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on the islet in human pancreas. The section was incubated with ab283265 overnight at 4℃. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

Heat mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0)

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Chromogranin A antibody [RM1025] (AB283265)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SH-SY5Y (human neuroblastoma cell line from bone marrow) cells labelling Chromogranin A with ab283265 at 1/50 dilution (10.74 μg/ml), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (2 μg/ml) (Green). Confocal image showing cytoplasmic staining in subsets of SH-SY5Y cells.

Negative control : Jurkat.

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (2.5 μg/ml) (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed.

• IP

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Immunoprecipitation - Anti-Chromogranin A antibody [RM1025] (AB283265)

Chromogranin A was immunoprecipitated from 0.35 mg SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate with ab283265 at 1/30 dilution (2 μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283265 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate 10ug

Lane 2 : ab283265 IP in SH-SY5Y whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab283265 in SH-SY5Y whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 30 seconds

Lower bands could be cleavage fragment. (PMID : 17991725)

All lanes:

Immunoprecipitation - Anti-Chromogranin A antibody [RM1025] (ab283265)

Predicted band size: 50 kDa

Observed band size: 75 kDa

false

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-Chromogranin A antibody [RM1025] (AB283265)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse colon (fresh) tissue labeling Chromogranin A with ab283265 at 1/500 dilution (1.074 μg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (2 μg/ml) (Green). Positive satining on mouse colon is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chromogranin A antibody [RM1025] (AB283265)

Immunohistochemical analysis of paraffin-embedded rat adrenal gland tissue labelling Chromogranin A with ab283265 at 1/5000 dilution (0.107 μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining in the rat adrenal gland. The section was incubated with ab283265 overnight at 4℃. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

Heat mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chromogranin A antibody [RM1025] (AB283265)

Immunohistochemical analysis of paraffin-embedded rat pancreas tissue labelling Chromogranin A with ab283265 at 1/5000 dilution (0.107 μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on the islet in rat pancreas. The section was incubated with ab283265 overnight at 4℃. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

Heat mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chromogranin A antibody [RM1025] (AB283265)

Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue labelling Chromogranin A with ab283265 at 1/5000 dilution (0.107 μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on the islet in mouse pancreas. The section was incubated with ab283265 overnight at 4℃. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

Heat mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0)

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Chromogranin A antibody [RM1025] (AB283265)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat colon (fresh) tissue labeling Chromogranin A with ab283265 at 1/500 dilution (1.074 μg/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (2 μg/ml) (Green). Positive staining on rat colon is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Chromogranin A antibody [RM1025] (AB283265)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Neuro-2a (mouse neuroblastoma cell line) cells labelling Chromogranin A with ab283265 at 1/50 dilution (10.74 μg/ml), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (2 μg/ml) (Green). Confocal image showing cytoplasmic staining in of Neuro-2a cells.

Negative control : NIH/3T3 (PMID : 7615829).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (2.5 μg/ml) (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chromogranin A antibody [RM1025] (AB283265)

Immunohistochemical analysis of paraffin-embedded mouse adrenal gland tissue labelling Chromogranin A with ab283265 at 1/5000 dilution (0.107 μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining in the mouse adrenal gland. The section was incubated with ab283265 overnight at 4℃. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

Heat mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0)

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Chromogranin A antibody [RM1025] (AB283265)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast / Left) and Neuro-2a (Mouse neuroblastoma neuroblast / Right) cells labelling Chromogranin A with ab283265 at 1/500 dilution (0.1 μg / Red) compared with a Rabbit monoclonal IgG (ab172730 / Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

Negative control : NIH/3T3. (PMID : 7615829)

• IP

Supplier Data

Immunoprecipitation - Anti-Chromogranin A antibody [RM1025] (AB283265)

Chromogranin A was immunoprecipitated from 0.35 mg Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysate with ab283265 at 1/30 dilution (2 μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283265 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysate 10ug

Lane 2 : ab283265 IP in Neuro-2a whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab283265 in Neuro-2a whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 3 min

Lower bands could be cleavage fragment. (PMID : 17991725)

All lanes:

Immunoprecipitation - Anti-Chromogranin A antibody [RM1025] (ab283265)

Predicted band size: 50 kDa

Observed band size: 75 kDa

false

• WB

Lab

Western blot - Anti-Chromogranin A antibody [RM1025] (AB283265)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Negative control : Jurkat.

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

This blot was developed using a higher sensitivity ECL substrate.

Exposure time : 3 min

All lanes:

Western blot - Anti-Chromogranin A antibody [RM1025] (ab283265) at 1/1000 dilution

Lane 1:

Human adrenal gland lysate at 20 µg

Lane 2:

SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate at 20 µg

Lane 4:

Neuro-2a (Mouse neuroblastoma neuroblast) whole cell 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

false

• WB

Lab

Western blot - Anti-Chromogranin A antibody [RM1025] (AB283265)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Negative control : NIH/3T3. (PMID : 7615829)

Exposure time : Lane 1-2 : 5 seconds Lane 3-5 : 92 seconds

All lanes:

Western blot - Anti-Chromogranin A antibody [RM1025] (ab283265) at 1/1000 dilution

Lane 1:

Mouse adrenal gland lysate at 20 µg

Lane 2:

Rat adrenal gland lysate at 20 µg

Lane 3:

Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 4:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 5:

PC-12 (Rat adrenal gland pheochromocytoma) whole cell 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

false

• ELISA

Supplier Data

ELISA - Anti-Chromogranin A antibody [RM1025] (AB283265)

ELISA using ab283265 at varying antibody concentrations and antigen concentration at 1000 ng/ml. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) (1/2500) was used as the secondary antibody.