41-9749-82

antibody from Invitrogen Antibodies

Targeting: APP AD1

Provider product page for 41-9749-82

Immunocytochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [3]
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Validations
Immunocytochemistry [1]
Other assay [4]

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Product number: 41-9749-82 - Provider product page
Provider: Invitrogen Antibodies
Product name: APP (Amyloid Precursor Protein) Monoclonal Antibody (22C11), eFluor™ 570, eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: Description: The monoclonal antibody 22C11 recognizes human, mouse, and rat APP (Amyloid Precursor Protein). APP is expressed in high abundance in the central nervous system and has three major isoforms resulting from alternative splicing. APP plays a role in synaptic formation and repair, anterograde neuronal transport, iron export, and hormonal regulation. Secreted APP (sAPP) may have neuroprotective effects against neurotoxic insult, oxidative stress, and excitotoxicity. APP belongs to a family that contains at least two homologs, amyloid precursor-like proteins 1 and 2 (APLP1 and APLP2). Similarities between APP and APLP, especially APLP2, suggest that APLP could share and compensate for the function of APP. Proteolytic cleavage of APP results in the generation of beta amyloid, which is the primary component of senile plaques. Senile plaques are one of the major histopathologic features of Alzheimer's disease. Abnormal regulation and processing of APP also plays a role in Down's syndrome, early onset familial Alzheimer's disease, cerebral hemorrhage, and arthritis.
Antibody clone number: 22C11
Concentration: 0.2 mg/mL

APP protein structure - 41-9749-82 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 3 Increased Abeta accumulation in iPSC-derived cerebral organoids from AD patients. Lysates of 4-5 cerebral organoids per iPSC line were analyzed by ELISA and western blotting at week 12. a - c Amounts of Abeta40 ( a ; APOE4: p = 0.2167, AD: p = 0.0002, APOE4 x AD: p = 0.4849, Con-E3 vs. AD-E3: p = 0.0287, Con-E3 vs. AD-E4: p = 0.0028, Con-E4 vs. AD-E3: p = 0.0171, Con-E4 vs. AD-E4: p = 0.0011) and Abeta42 ( b ; APOE4: p = 0.1014, AD: p < 0.0001, APOE4 x AD: p = 0.7778, Con-E3 vs. AD-E3: p < 0.0001, Con-E3 vs. AD-E4: p < 0.0001, Con-E4 vs. AD-E3: p < 0.0001, Con-E4 vs. AD-E4: p < 0.0001) in the RIPA fraction were measured by ELISA. Data were normalized to the total protein concentration of the respective sample. The ratio of Abeta42/Abeta40 was calculated accordingly ( c ; APOE4: p = 0.9549, AD: p = 0.0034, APOE4 x AD: p = 0.5331, Con-E4 vs. AD-E3: p = 0.0397). d - f Amounts of sAPPalpha ( d ; APOE4: p = 0.1081, AD: p = 0.8009, APOE4 x AD: p = 0.1351), sAPPbeta (E; APOE4: p = 0.7772, AD: p = 0.4410, APOE4 x AD: p = 0.0936) and CTF-beta ( f ; APOE4: p = 0.1150, AD: p = 0.5326, APOE4 x AD: p = 0.9877) in RIPA were measured by ELISA. Data are shown as ratios to Con-E3 after normalization to total protein concentration. g , h Amounts of full-length APP were assessed by western blotting analysis using 22C11 monoclonal antibody in RIPA lysate ( APOE4: p = 0.0331, AD: p = 0.6706, APOE4 x AD: p = 0.3170). All data are expressed as mean +- SEM ( N = 5). ANCOVA for APOE4 , AD stat

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2. GSAP interacts with Fe65 to regulate APP phosphorylation and trafficking. (A) Immunoblot analysis of protein levels in N2a695 cells transfected with control or GSAP siRNA (left panel). Quantification of APP phosphorylation at Thr668 normalized to total APP level (right panel). Data represent mean +- SEM; unpaired t test, **, P < 0.01. pT668, phospho-Thr668. Representative data of four experiments. (B) Co-IP analysis of full-length GSAP (HA-tagged) interaction with full-length Fe65 (Flag-tagged) using Flag antibody in HEK293T cells. Representative data of two experiments. (C) Co-IP analysis of GSAP C-terminal 16K domain (HA-tagged) coprecipitation with full-length Fe65 (Flag-tagged) using a Flag antibody in HEK293T cells. Representative data of two experiments. (D) Co-IP analysis of endogenous Fe65 interaction with GSAP and PP1 using Fe65 antibody in HEK293T cells. GSAP was detected using an antibody from R&D Systems. Representative data of two experiments. (E) Genomic DNA from CAD WT and Fe65KO cells was isolated, and PCR-amplified fragments flanking the CRISPR-Cas9 cleavage site were generated. PCR fragments were cloned into TOPO vector for Sanger sequencing. A 1-bp insertion (red) and deletion (blue) was identified in Fe65KO CAD cells (upper panel). Immunoblot analysis of proteins from WT and Fe65KO CAD cells (lower panel). (F) Immunoblot analysis of protein levels in CAD cells transiently overexpressing APP (left panel). Quantification of APP phosphorylation at T

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: FIGURE 3 miR-425 deficiency activates amyloidogenic APP processing and systematic neuroinflammation. (a) Generation of Mir425 +/- mice with CRISPR/Cas9 and genotypes verification of Mir425 +/- and WT mice using PCR. (b) In situ hybridization of miR-425 in hippocampus and cortex of Mir425 +/- and WT mice. Scale bars, 50 mum. (c, d) RT-PCR of miR-425 level in the hippocampus of Mir425 +/- and WT mice at 3 months and 15 month. (e) Western blot of APP, BACE1, sAPPalpha/beta, CTF-alpha, and CTF-beta in the brain of miR-425 +/- and WT mice. (f) ELISA of Abeta40 and CTF-beta in the brain of 15-month-old Mir425 +/- and WT mice. (g, h) Western blot of APP and CTF-beta in PC12 cells transfected with Antagomir-425 and Agomir-425. (i, j) Luciferase assay of miR-425 and APP 3'UTR. (k, l) Immunofluorescence and quantification of GFAP-positive and Iba1-positive glial cells in the hippocampus of 15-month-old Mir425 +/- and WT mice. Scale bars, 50 mum. (m, n) Microarray cytokines assay of hippocampal cell lysates in 15-month-old Mir425 +/- and WT mice. Data were presented as mean +- SEM. Two-tailed unpaired Student's t test. *** p < 0.001; ** p < 0.01; * p < 0.05

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 7 AgomiR-425 oligonucleotide treatment ameliorates pathological and behavioral phenotypes in APP/PS1 mice. (a) Stereotactic injection of FAM-labeled oligonucleotide in the hippocampus of 6-month-old APP/PS1 mice. Scale bars, 200 mum (left), 20 mum (right). (b) Chromogenic in situ hybridization and quantification of miR-425 in hippocampus of APP/PS1 mice. Scale bars, 100 mum, n = 6 per group. (c) Immunofluorescence and quantification of APP and PTEN in AgomiR-425- and scramble-treated APP/PS1 mice, n = 6 per group. Scale bars, 50 mum. (d) Immunofluorescence and quantification of amyloid plaques in AgomiR-425- and scramble-treated APP/PS1 mice, n = 6 per group. Scale bars, 100 mum. (e, f) Immunofluorescence and quantification of APP-CTFs, GFAP and Iba1 in AgomiR-425- and scramble-treated APP/PS1 mice. Scale bars, 25 mum. (g) Immunofluorescence and quantification of Ki67- and DCX-positive neurons in AgomiR-425-and scramble-treated APP/PS1 mice. Scale bars, 50 mum. (h-j) MWM data showing times across the target, distance in target quadrant for four times and swimming speed of APP/PS1 mice, n = 12 per group. (k, l) NOR tests showing the total distance of novel object exploration and preference index of APP/PS1 mice, n = 12 per group. (m, n) Y-maze tests showing alternation percentage and the number of arm entered of APP/PS1 mice, n = 12 per group. Data were presented as mean +- SEM. Two-tailed unpaired Student's t test. *** p < 0.001; * p < 0.05