PA5-19431

antibody from Invitrogen Antibodies

Targeting: HNRNPA1 ALS20, hnRNP-A1, HNRPA1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Immunohistochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-19431

Provider

Invitrogen Antibodies

Product name

hnRNP A1 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

Heat mediated antigen retrieval recommended prior to tissue staining. This antibody is predicted to react with mouse, rat and macaque monkey based on sequence homology. Store antibody at 4ºC for 1-2 weeks. For long-term storage, store at -20ºC.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

100 µg

Concentration

0.9 mg/mL

Storage

-20°C or -80°C if preferred

References

Submitted references

Structural basis for terminal loop recognition and stimulation of pri-miRNA-18a processing by hnRNP A1.

Kooshapur H, Choudhury NR, Simon B, Mühlbauer M, Jussupow A, Fernandez N, Jones AN, Dallmann A, Gabel F, Camilloni C, Michlewski G, Caceres JF, Sattler M
Nature communications 2018 Jun 26;9(1):2479

---

Genetic variation and RNA structure regulate microRNA biogenesis.

Fernandez N, Cordiner RA, Young RS, Hug N, Macias S, Cáceres JF
Nature communications 2017 May 3;8:15114

---

SAF-A Regulates Interphase Chromosome Structure through Oligomerization with Chromatin-Associated RNAs.

Nozawa RS, Boteva L, Soares DC, Naughton C, Dun AR, Buckle A, Ramsahoye B, Bruton PC, Saleeb RS, Arnedo M, Hill B, Duncan RR, Maciver SK, Gilbert N
Cell 2017 Jun 15;169(7):1214-1227.e18

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of HepG2 whole cell lysate using Product # PA5-19431, hnRNP A1 primary antibody at a dilution of 1 µg/mL (lane 1). Staining of MCF7 whole cell lysate at a dilution of 1 µg/mL (Lane 2). Staining of SHSY-5Y whole cell lysate at a dilution of 1 µg/mL (lane 3). Blot treated with a secondary IR Dye680-conjugated Goat polyclonal anti-Rabbit antibody was used at a dilution of 1:10000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts (30 µg lysate) of HepG2 (Lane 1), MCF7 (Lane 2), MDA-MB-231 (Lane 3), A549 (Lane 4) and A-431 (Lane 5). The blot was probed with hnRNP A1 Polyclonal Antibody (Product # PA5-19431, 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 band at ~38 kDa corresponding to hnRNP A1 was observed across cell lines tested.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of hnRNP A1 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with hnRNP A1 Rabbit Polyclonal Antibody(Product # PA5-19431) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nucleus localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical (formalin-fixed, paraffin-embedded) staining of human skin tissue tissue using Product # PA5-19431, anti-hnRNP A1 antibody. Primary antibody was used at a concentration of 5 µg/mL and exposed for 15 mins at room temp. The sample was pretreated using heat mediated antigen retrieval with Sodium Citrate Buffer (pH6/20mins). The detection method was a HRP conjugated polymer, DAB chromogen and the sample was counterstained with haematoxylin and mounted with DPX.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical (formalin-fixed, paraffin-embedded) staining of human skin tissue tissue using Product # PA5-19431, anti-hnRNP A1 antibody. Primary antibody was used at a concentration of 5 µg/mL and exposed for 15 mins at room temp. The sample was pretreated using heat mediated antigen retrieval with Sodium Citrate Buffer (pH6/20mins). The detection method was a HRP conjugated polymer, DAB chromogen and the sample was counterstained with haematoxylin and mounted with DPX.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Detection of binding of endogenous hnRNP A1 protein to specific RNA using Anti-hnRNP A1 Antibody: RNA Immunoprecipitation (RIP) was performed using Anti-hnRNP A1 Rabbit Polyclonal Antibody (Product # PA5-19431, 5 µg) on clarified whole cell lysate from 4 million HCT 116 cells. Normal Rabbit IgG was used as a negative IP control. Immunoprecipitated RNA was purified by RiboPure™ RNA Purification Kit (Product # AM1924) and analyzed by RT-PCR using the Power SYBR® Green RNA-to-CT™ 1-Step Kit (Product # 4389986) with primer pairs over ATF3, CYR61 (positive) and 18S rRNA (negative). Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.