MA5-13096

antibody from Invitrogen Antibodies

Targeting: CHGA

Provider product page for MA5-13096

Western blot

Immunocytochemistry

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [41]
Comments [0]
Validations
Western blot [3]
Immunocytochemistry [3]
Immunohistochemistry [5]
Other assay [3]

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Product number: MA5-13096 - Provider product page
Provider: Invitrogen Antibodies
Product name: Chromogranin A Monoclonal Antibody (LK2H10)
Antibody type: Monoclonal
Antigen: Other
Description: MA5-13096 targets Chromogranin A in IHC (P), ICC/IF and WB applications and shows reactivity with Human, Non-human primate, Porcine, Rabbit, and Rat samples. The MA5-13096 immunogen is human pheochromocytoma.
Reactivity: Human, Rat, Porcine, Rabbit
Host: Mouse
Isotype: IgG
Antibody clone number: LK2H10
Vial size: 500 µL
Concentration: 0.2 mg/mL
Storage: 4° C

CHGA protein structure - MA5-13096 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot of Chromogranin A using Chromogranin A Monoclonal Antibody (Product # MA5-13096) on PC12 Cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of Chromogranin A was achieved by transfecting SH-SY5Y cells with Chromogranin A specific siRNAs (Silencer® select Product # s2992, s2993) and further treating with PTI (1X for 4hr). Western blot analysis (Fig. a) was performed using modified whole cell extracts (1% SDS) from Chromogranin A knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with Chromogranin A Monoclonal Antibody (LK2H10) (Product # MA5-13096, 1ug/ml) and Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Chromogranin A.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-Chromogranin A Monoclonal Antibody (Product # MA5-13096) and a ~60 kDa band corresponding to Chromogranin A was observed in S-HSY5Y cell line treated with Protein Transport Inhibitor (PTI, 1X for 4r) and FL1-06 (10uM for 4hr). An uncharacterized band at ~70kDa was also observed across the panel tested. Whole cell extracts (30ug) of SH-SY5Y (Lane 1), SH-SY5Y treated with PTI (1X for 4hr) (Lane 2) and SH-SY5Y treated with FL1-06 (10uM for 4hr) (Lane 3) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1ug/ml) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Chromogranin A (green) showing staining in the cytoplasm of PC12 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Chromogranin A monoclonal antibody (Product # MA5-13096) in 3% BSA-PBS at a dilution of 1:50 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Chromogranin A (green) showing staining in the cytoplasm of SH-SY5Y cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Chromogranin A monoclonal antibody (Product # MA5-13096) in 3% BSA-PBS at a dilution of 1:50 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Chromogranin A (green) showing staining in the cytoplasm of U87-MG cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Chromogranin A monoclonal antibody (Product # MA5-13096) in 3% BSA-PBS at a dilution of 1:50 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Formalin-fixed, paraffin-embedded human pancreatic tumor stained with Chromogranin A antibody using peroxidase-conjugate and AEC chromogen. Note cytoplasmic staining of tumor cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Formalin-fixed, paraffin-embedded rat pancreas stained with Chromogranin A antibody using peroxidase-conjugate and AEC chromogen. Note cytoplasmic staining of islet cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Chromogranin A showing positive staining in the cytoplasm of paraffin-treated Human pancreas tissue (right) compared with a negative control in the absence of primary antibody (left). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Chromogranin A monoclonal antibody (Product # MA5-13096) diluted by 3% BSA-PBS at a dilution of 1:2000 overnight at 4°C in a humidified chamber. Tissues were washed extensively PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Chromogranin A showing positive staining in the cytoplasm of paraffin-treated Human small intestine (right) compared with a negative control in the absence of primary antibody (left). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Chromogranin A monoclonal antibody (Product # MA5-13096) diluted by 3% BSA-PBS at a dilution of 1:2000 overnight at 4°C in a humidified chamber. Tissues were washed extensively PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Chromogranin A showing positive staining in the cytoplasm of paraffin-treated Rat pancreas tissue (right) compared with a negative control in the absence of primary antibody (left). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Chromogranin A monoclonal antibody (Product # MA5-13096) diluted by 3% BSA-PBS at a dilution of 1:1000 overnight at 4°C in a humidified chamber. Tissues were washed extensively PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Establishment of neuroendocrine neoplasm (NEN) patient-derived xenograft (PDX) models: ( A ) Tumor samples from NEN patients were cryopreserved, thawed, and injected into the flank of immunocompromised NOD Scid Gamma (NSG) mice. Two mice developed subcutaneous tumors at three months post injection (NEC913 and NEC1452 PDX models). Both PDX models have been passaged in 6 generations of mice. ( B ) Formalin-fixed and paraffin-embedded tumor sections are stained with H&E and stained for Ki67 and neuroendocrine tumor markers such chromogranin A (CgA), synaptophysin (SYP), and somatostatin receptor 2 (SSTR2) by IHC. Scale bar represents 40 mum. ( C ) Exome sequencing of NEC913 and NEC1452 PDX tumors demonstrated mutations in TP53 and RB1 .

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 7 Analysis of human primary, relapsed and bone metastatic PCa specimens for the expression of CD44 and Twist1. ( A ) IHC staining was performed on ten primary, ten relapsed and ten bone metastatic PCa specimens using a specific antibody against a NE marker CgA. Representative images of CgA expression are shown depicting negative staining in primary and positive staining in relapsed and bone metastatic PCa. ( B ) The RNA level of CD44 and Twist1 were measured in each specimen by RT-qPCR. The statistical significance was calculated between the three groups of specimens using Student's t-test . Statistical significance between groups is indicated. Data represent means +- SD. * p < 0.05 by Student's-t test.