- MA5-33117 - Invitrogen Antibodies ATF4 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot analysis of precipitated ATF4 from Hela whole cell lysate using a ATF4 monoclonal antibody (Product # MA5-33117). An HRP-conjugated Protein G antibody was used as the secondary antibody (1:2000). Lane 1: Rabbit control IgG. Lane 2: Hela whole cell lysate (500µg). Lane 3: Hela whole cell lysate (20µg).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of ATF4 using a ATF4 Monoclonal antibody (Product # MA5-33117) at a concentration of 1.6 µg/mL. Positive WB detected in: Hela whole cell lysate, MCF-7 whole cell lysate, 293 whole cell lysate, HepG2 whole cell lysate, Jurkat whole cell lysate, K562 whole cell lysate. A secondary Goat polyclonal antibody to rabbit IgG was applied at a 1:50,000 dilution. Observed band size: 50 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: CRISPR-Cas9 mediated genome editing ofATF4 (as confirmed by next generation sequencing) was achieved by using LentiArray™ Lentiviral sgRNA (Product # A32042, AssayID CRISPR988840_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Fig (a) Western blot analysis of ATF4 was performed by loading 30 µg of HeLa wild type (Lane 1), HeLa wild type treated with 2 µg/mL tunicamycin for 8hrs (Lane 2), HeLa Cas9 (Lane 3), HeLa Cas9 treated with 2 µg/mL tunicamycin for 8hrs (Lane 4), HeLa Cas9 cells transduced with ATF4 Lentiviral sgRNA (Lane 5) and HeLa Cas9 cells transduced with ATF4 Lentiviral sgRNA and treated with 2 µg/mL tunicamycin for 8hrs (Lane 6) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein 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 Anti-ATF4 Recombinant Rabbit Monoclonal Antibody (Product # MA5-33117, 1.6 µg/mL dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036 1:20000 dilution).Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). A reduced signal in sgRNA transduced cells using the LentiArray™ CRISPR product line confirms that antibody is specific toATF4. Uncharacterized band was obs

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of precipitated ATF4 from Hela whole cell lysate using a ATF4 monoclonal antibody (Product # MA5-33117). An HRP-conjugated Protein G antibody was used as the secondary antibody (1:2000). Lane 1: Rabbit control IgG. Lane 2: Hela whole cell lysate (500µg). Lane 3: Hela whole cell lysate (20µg).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of ATF4 using a ATF4 Monoclonal antibody (Product # MA5-33117) at a concentration of 1.6 µg/mL. Positive WB detected in: Hela whole cell lysate, MCF-7 whole cell lysate, 293 whole cell lysate, HepG2 whole cell lysate, Jurkat whole cell lysate, K562 whole cell lysate. A secondary Goat polyclonal antibody to rabbit IgG was applied at a 1:50,000 dilution. Observed band size: 50 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of ATF4 in HepG2 cells using a ATF4 monoclonal antibody (Product # MA5-33117) at a dilution of 1:53. The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated Goat Anti-Rabbit IgG (H+L). Cells were counter-stained with DAPI.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of ATF4 in HepG2 cells using a ATF4 monoclonal antibody (Product # MA5-33117) at a dilution of 1:53. The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated Goat Anti-Rabbit IgG (H+L). Cells were counter-stained with DAPI.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of ATF4 in paraffin embedded human breast cancer using a ATF4 monoclonal antibody (Product # MA5-33117) at a dilution of 1:160. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow cytometric analysis of ATF4 in Hela cells using a monoclonal antibody (Product # MA5-33117) at a dilution of 1:50. The cells were fixed with 70% Ethyl alcohol (18h) and then permeabilized with 0.3% Triton X-100 for 2 min. The cells were then incubated in 1x PBS /10% normal goat serum to block non-specific protein-protein interactions followed by primary antibody for 1 h at 4°C. The secondary antibody used was FITC goat anti-rabbit IgG (H+L) at 1:200 dilution for 1 h at 4°C. Control antibody (green line) was used under the same conditions. Acquisition of >10,000 events was performed.

MA5-33117

Antibody data