Ab21685

Manufactured by Merck Group

Ab21685 is a laboratory equipment product. It is designed to perform a core function, but a detailed description cannot be provided while maintaining an unbiased and factual approach without extrapolation.

Automatically generated - may contain errors

Lab products found in correlation

Ab12223, by Merck Group (1 mentions) Sc 376768, by Abcam (1 mentions) Sc 3887, by Santa Cruz Biotechnology (1 mentions) Alexa 594 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) Mouse anti myc, by Santa Cruz Biotechnology (1 mentions) Mouse anti sma, by Merck Group (1 mentions) Rabbit anti c myc, by Merck Group (1 mentions) 4 6 diamidino 2 phenylindole dapi staining reagents, by Thermo Fisher Scientific (1 mentions) Rabbit anti yap1, by Cell Signaling Technology (1 mentions) Alexa 488 phalloidin, by Thermo Fisher Scientific (1 mentions) Hrp labeled secondary antibody, by Santa Cruz Biotechnology (1 mentions) Ab2302, by Abcam (1 mentions) Ab2799, by Abcam (1 mentions) Ab53098, by Abcam (1 mentions) Ab104223, by Abcam (1 mentions) Ab144p, by Merck Group (1 mentions) Fluoview fv1000, by Olympus (1 mentions) Mab374, by Merck Group (1 mentions)

4 protocols using ab21685

Validation of Anti-GRP78 Antibodies

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Seven commercial anti-GRP78 Abs (Santa Cruz Biotechnology, sc-376768, sc-1051, sc-1050, and sc-13968; Abcam, ab21685, ab12223; Sigma-Aldrich, G8918) were tested. Control IgGs were goat, rabbit, and mouse polyclonal Abs (Santa Cruz Biotechnology, sc-3887, sc-2027, and sc-2025). Cultured cells were incubated with these Abs or IgGs (10, 20, 40, and 80 μg/ml) for 1 hour and fixed with 4% PFA.

Shimizu F., Schaller K.L., Owens G.P., Cotleur A.C., Kellner D., Takeshita Y., Obermeier B., Kryzer T.J., Sano Y., Kanda T., Lennon V.A., Ransohoff R.M, & Bennett J.L. (2017). Glucose-regulated protein 78 autoantibody associates with blood-brain barrier disruption in neuromyelitis optica. Science translational medicine, 9(397), eaai9111.

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Comprehensive Antibody Characterization for Cell-Based Studies

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The following primary antibodies were used: rabbit anti-NOS1AP (Novus Biologicals, NBP2-38758 and NBP2-38151), guinea anti-nephrin (Progen, GP-N2), mouse anti-SMA (Sigma-Aldrich, A2547), mouse anti-CD31 (MA3100, Thermo Fisher Scientific), mouse anti-NWASP (LSBio, LS-C133098-100), mouse anti-DIAPH3 (Proteintech, 14342-1-AP), rabbit anti-c-Myc (Sigma-Aldrich, C3956), mouse anti-myc (Santa Cruz Biotechnology, SC-40), mouse horseradish peroxidase (HRP)–linked anti–β-actin (Abcam, ab20272), rabbit anti-YAP1 (Cell Signaling Technology, 4912), anti-Golgin B1 (GOLGB1) (Sigma-Aldrich, HPA011008), anti-GRP78 Binding Immunoglobulin Protein (BiP) (Abcam, ab21685), anti-Nucleoporin 153 (NUP153) (Sigma-Aldrich, HPA027897), and anti–cleaved CASP3 (Abcam, ab2302). Donkey anti-mouse, anti-guinea, and anti-rabbit Alexa 488– and Alexa 594–conjugated secondary antibodies; 4′,6-diamidino-2-phenylindole (DAPI) staining reagents; and phalloidin–Alexa 488 were obtained from Invitrogen (Thermo Fisher Scientific). HRP-labeled secondary antibodies were purchased from Santa Cruz Biotechnology.

Majmundar A.J., Buerger F., Forbes T.A., Klämbt V., Schneider R., Deutsch K., Kitzler T.M., Howden S.E., Scurr M., Tan K.S., Krzeminski M., Widmeier E., Braun D.A., Lai E., Ullah I., Amar A., Kolb A., Eddy K., Chen C.H., Salmanullah D., Dai R., Nakayama M., Ottlewski I., Kolvenbach C.M., Onuchic-Whitford A.C., Mao Y., Mann N., Nabhan M.M., Rosen S., Forman-Kay J.D., Soliman N.A., Heilos A., Kain R., Aufricht C., Mane S., Lifton R.P., Shril S., Little M.H, & Hildebrandt F. (2021). Recessive NOS1AP variants impair actin remodeling and cause glomerulopathy in humans and mice. Science Advances, 7(1), eabe1386.

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Immunocytochemistry of iMN Markers

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iMNs plated on coverslips were fixed using 4% paraformaldehyde (PFA) for 15 min and blocked for 1 h with 3% bovine serum albumin (BSA) and 0.1% TritonX-100 in phosphate buffered saline PBS. iMNs were incubated with the following primary antibody: mouse anti-GRP75 (Abcam, ab2799, 1:200), rabbit anti-GRP75 (1:200, Abcam, ab53098), rabbit anti-BiP (Abcam, ab21685, 1:500), goat anti-ChAT (Millipore, AB144P, 1:500), chicken anti-MAP2 (Sigma Aldrich, AB15452, 1:500), rat anti Isl1-2 (DSHB 39.4D5), mouse anti-HB9 (1:500, DSHB 81.5C10), mouse anti-TDP43 (ABCAM ab104223, 1:500), in blocking buffer overnight at 4 °C. After three washes with PBS, cells were incubated in blocking buffer with appropriate Alexa Fluor secondary antibodies and DAPI for 1 h at RT, followed by three PBS washes and coverslips were mounted on glass slides. Confocal images were acquired with FluoViewTM FV1000 (Olympus) fitted with a 20 × or 40 × air objective and 60 × immersion oil objective.

Pilotto F., Schmitz A., Maharjan N., Diab R., Odriozola A., Tripathi P., Yamoah A., Scheidegger O., Oestmann A., Dennys C.N., Sinha Ray S., Rodrigo R., Kolb S., Aronica E., Di Santo S., Widmer H.R., Charlet-Berguerand N., Selvaraj B.T., Chandran S., Meyer K., Zuber B., Goswami A., Weis J, & Saxena S. (2022). PolyGA targets the ER stress-adaptive response by impairing GRP75 function at the MAM in C9ORF72-ALS/FTD. Acta Neuropathologica, 144(5), 939-966.

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Western Blot Analysis of Cellular Proteins

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Cell pellets were lysed with either RIPA (Thermo Scientific Cat#89900) (with sonication) or IP (without sonication) lysis buffer. 50 μg of cell lysate was loaded onto a BIS‐Tris PAGE gel and run at 120 volts for 1 h. Protein was transferred onto a PDVF membrane and blocked for 1 h at room temperature using Odyssey blocking buffer. Antibodies against either SOD1 (Cell signaling cat# 2770S), BIP (Abcam Cat# ab21685), and GAPDH (Millipore cat# MAB374) were incubated overnight at 4°C. Following three TBS‐T washes cells were then incubated with 1/15,000 Licor secondaries (goat anti Mouse 680 red, cat# 926‐68,020, goat anti rabbit 800 green, cat# 926‐32211, goat anti rabbit 680 red, cat# 926‐68071, goat anti mouse 800 green, cat# 926‐32210) for 1 h at room temperature and washed again three times.

Dennys C.N., Roussel F., Rodrigo R., Zhang X., Sierra Delgado A., Hartlaub A., Saelim‐Ector A., Ray W., Heintzman S., Fox A., Kolb S.J., Beckman J., Franco M.C, & Meyer K. (2022). CuATSM effectively ameliorates ALS patient astrocyte‐mediated motor neuron toxicity in human in vitro models of amyotrophic lateral sclerosis. Glia, 71(2), 350-365.

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