MA1-19287

antibody from Invitrogen Antibodies

Targeting: PAG1 CBP, PAG

Provider product page for MA1-19287

Western blot

Immunoprecipitation

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [2]
Comments [0]
Validations
Other assay [3]

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Product number: MA1-19287 - Provider product page
Provider: Invitrogen Antibodies
Product name: PAG1 Monoclonal Antibody (PAG-C1)
Antibody type: Monoclonal
Antigen: Synthetic peptide
Description: This antibody recognizes an epitope located in the intracellular C-terminal domain of Csk-binding protein (Cbp / PAG), a 46 kDa ubiquitously expressed transmembrane adaptor protein present in membrane rafts (glycosphingolipid-enriched microdomains), which however migrates on SDS PAGE gels anomalously as an 80 kDa molecule. A suggested positive control for Immunoprecipitation is RAJI human Burkitt lymphoma cell line. A suggested positive control for immunohistochemistry is appendix (germinal center of lymphatic follicle). For IHC staining, heat-mediated antigen retrieval in citrate buffer, pH 6.1, is recommended.
Reactivity: Human, Mouse, Rat, Bovine
Host: Mouse
Isotype: IgG
Antibody clone number: PAG-C1
Vial size: 100 µg
Concentration: 1 mg/mL
Storage: 4° C, do not freeze

PAG1 protein structure - MA1-19287 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunoprecipitation of PAG1 using a monoclonal antibody (Product # MA1-19287).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunoprecipitation of human PAG/Cbp from the lysate of RAJI human Burkitt lymphoma cell line. Western blot was immunostained with anti-human PAG (MEM-255). Note: PAG/Cbp is a 46 kDa adaptor protein, which however migrates on SDS PAGE gels anomalously as an 80 kDa molecule. Lane 1,2: immunoprecipitation with anti-PAG (PAG-C1) Monoclonal antibody (Product # MA1-19287); Lane 3: immunoprecipitation with anti-PAG (polyclonal antibody).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 1: SH-SY5Y cells expressing cytosolic PAG1 (PAG1 TM- ) were generated using CRISPR/Cas9. (A) WT PAG1 has a small extracellular N-terminal region, transmembrane domain (TM), and long intracellular C-terminal tail. Several phosphorylated residues and proline-rich regions (PRR, PXXP motif, green) act as docking sites for protein-protein interactions. Phosphorylated Y163/Y181 and Y387/417 (blue) are binding sites for FYN and LYN SH2 domains, respectively (; , ; ). (B) CRISPR sgRNA recognition site is highlighted in green. Sequencing results indicated a single base pair insertion that resulted in a stop codon to generate a heterozygous PAG1 - SH-SY5Y cell line (red). (C) Met residues upstream of the detected peptides act as alternate translation start sites to produce the cytosolic PAG1 product (PAG1 TM- ). PAG1 peptides were detected via mass spectrometry on PAG1 - cell lines (amino acid numbers are indicated in black). (D) Organelle fractionation comparing the endosomal distribution of WT PAG1 vs. PAG1 TM- . Fractions were taken from 25% to 2.5% iodixanol gradients and decrease in density from left to right. L, biotinylated ladder. PAG1 is extensively posttranslationally modified in WT cells and displays multiple bands on Western blots; in PAG1 TM- , a single band is visible, suggesting few such modifications. (E) Levels of activated SFKs were increased in PAG1 TM- -expressing SH-SY5Ys (blue) compared with WT SH-SY5Y cells (red). Cells were stained with a pSFK antibody re