MA5-14820

antibody from Invitrogen Antibodies

Targeting: DNAJC3 ERdj6, HP58, P58, p58(IPK), P58IPK, PRKRI

Western blot (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-14820

Provider

Invitrogen Antibodies

Product name

DNAJC3 Monoclonal Antibody (J.98.6)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

It is not recommended to aliquot this antibody.

Reactivity

Human, Mouse

Host

Rabbit

Isotype

IgG

Antibody clone number

J.98.6

Vial size

100 µL

Storage

-20°C

References

Submitted references

Inhibition of eIF2α Dephosphorylation Protects Hepatocytes from Apoptosis by Alleviating ER Stress in Acute Liver Injury.

Tang YJ, Chen H, Yi Y, Chen GM, Yang FW, Li Y, Tian RD, Huang WG, Cheng QJ, He YH
BioMed research international 2020;2020:2626090

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Inhibiting alpha subunit of eukaryotic initiation factor 2 dephosphorylation protects injured hepatocytes and reduces hepatocyte proliferation in acute liver injury.

Chen G, Yang X, He Y, Tang Y, Tian R, Huang W, Chen H, Yang F, Li Y, Lin S
Croatian medical journal 2019 Dec 31;60(6):532-544

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of p58IPK in extracts from Beta-TC-6 and NIT-1 cells using p58IPK monoclonal antibody (Product # MA5-14820).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of DNAJC3 was achieved by transfecting HeLa cells with DNAJC3 specific siRNAs (Silencer® select Product # s11190, s11188). Western blot analysis (Fig. a) was performed using membrane enriched extracts from the DNAJC3 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Anti-DNAJC3 Monoclonal Antibody (Product # MA5-14820, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 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 DNAJC3.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts (30 µg lysate) of HeLa (Lane 1), Hep G2 (Lane 2), Raji (Lane 3), NIH/3T3 (Lane 4), MOLT-4 (Lane 5), COLO 205 (Lane 6), tissue extracts of Mouse Pancreas (Lane 7), Mouse Stomach (Lane 8) and Mouse Colon (Lane 9). The blot was probed with Anti-DNAJC3 Monoclonal Antibody (Product # MA5-14820, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 58 kDa band corresponding to DNAJC3 was detected across all the cell lines and tissues tested.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 3 The inhibition of alpha subunit of eukaryotic initiation factor 2 (eIF2alpha) dephosphorylation mitigates carbon tetrachloride (CCl 4 )-induced hepatocyte proliferation in mice. Male BALB/c mice were pretreated with vehicle (dimethyl sulfoxide + phosphate buffer solution), salubrinal, integrated stress response inhibitor (ISRIB) for 2 h, or recombinant AAV8 expressing DnaJC3 for 4 weeks, then injected with olive oil or CCl 4 . Relative levels of intrahepatic cyclin D1 and proliferating cell nuclear antigen (PCNA) expression at 24 h after CCl 4 injection were determined by Western blot. PCNA expression was assessed by immunohistochemistry. ( A ) Salubrinal, ISRIB, or overexpressed DnaJC3 altered eIF2alpha phosphorylation, DnaJC3, and PCNA expression (n = 6; total mice = 30). ( B ) Salubrinal, ISRIB, or ( C ) overexpressed DnaJC3 altered the levels of CCl 4 -induced eIF2alpha phosphorylation, cyclin D1, and PCNA expression (n = 8, total mice = 40, day 1). ( D ) Immunohistochemistry analysis of hepatic PCNA expression (magnification x100).