Chloride Channel, Nucleotide-Sensitive, 1A (CLNS1A) antibody

Details for Product No. ABIN4903315, Supplier: Log in to see
Antigen
  • CLCI
  • CLNS1B
  • ICln
  • 2610036D06Rik
  • 2610100O04Rik
  • Clci
  • Clcni
  • ICLN
  • Icln
  • pICln
  • clci
  • clns1b
  • icln
  • pIcIn
  • chloride nucleotide-sensitive channel 1A
  • chloride channel, nucleotide-sensitive, 1A
  • chloride channel, nucleotide-sensitive, 1A L homeolog
  • CLNS1A
  • Clns1a
  • clns1a.L
  • clns1a
Alternatives
anti-Human Chloride Channel, Nucleotide-Sensitive, 1A antibody for Western Blotting
Reactivity
Human
58
21
19
7
5
2
2
2
1
1
1
1
1
1
Host
Rabbit
55
4
Clonality
Polyclonal
Conjugate
Un-conjugated
1
1
1
1
1
1
1
1
1
1
1
1
1
1
Application
Flow Cytometry (FACS), Immunohistochemistry (IHC), Western Blotting (WB)
46
14
13
13
12
11
7
5
4
3
3
Options
Supplier
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Supplier Product No.
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'Independent Validation' Badge
Antigen CLNS1A
Lot Number 03R181011
Method validated Western Blotting
Positive Control

HeLa cells

Independent antibody

Negative Control

HeLa cells, CLNS1A knockdown using Dharmacon smartpool siRNA

Notes

Passed. ABIN4903315 specifically reveals endogenous CLNS1A in HeLa cell lysates while no signal is detected subsequently to knock-down of the antigen.

Primary Antibody ABIN4903315
Secondary Antibody HRP-conjugated goat anti-rabbit IgG (H+L) (Jackson ImmunoResearch, 111-035-003)
Protocol
  • Seed 3x104 HeLa cells in DMEM (PAN Biotech, P04-41450, 2730618) supplemented with 10% FBS (PAN Biotech, P03-3302), without antibiotics, in 6 well plates in 2ml media.
  • Grow cells for 24h at 37°C and 5% CO2 to 70% confluency.
  • Change medium.
  • Transfect cells with 60pmol ON-TARGETplus SmartPool siRNA human (Dharmacon, L-012571-00-0005, lot 170831) in 125µl 150mM NaCl added to 7.5µl Lipofectamine RNAiMax (13778030, lot 1911816) in 125µl 150mM NaCl.
  • Grow cells for 24h at 37°C and 5% CO2.
  • Change medium.
  • Grow cells for 72h at 37°C and 5% CO2.
  • Wash cells once with PBS (PAN Biotech, P04-36500, lot 3620618).
  • Detach cells with 100µl trypsin per well.
  • For WB the cells were once washed with PBS (PAN Biotech) before treated with 100µl of Trypsin (PAN Biotech, P10-023100, lot 6860417) per well.
  • Once cells are detached add 900µl medium.
  • Count cells.
  • Add 40µl 1x Laemmli buffer (Roth, K929.1, lot 227259243) to 2x105 cells. Heat up to 99°C for 10min.
  • Separate 10µl of the lysates on a freshly cast denaturing 12% SDS-PAGE gel in an electrophoresis chamber for 1h at 120V.
  • Transfer proteins onto PVDF membrane (Merck, IPVH0010, lot K5KN7128I) with a wet blotting system for 1h at 80V.
  • Block the membrane with 1x Roti Block (Roth, A151.2, lot 148270222) for 30min at RT.
  • Incubation with primary
    • rabbit anti-CLNS1A antibody (antibodies-online, ABIN4903315, lot 03R181011) diluted 1:500 in 1x Roti Block for 1h at RT,
    • mouse anti-CLNS1A antibody (Santa Cruz, sc-271454, G3010) diluted 1:3000 in 1x Roti Block for 1h at RT, or
    • rat anti-tubulin antibody (YOL 1/34 gifted) diluted 1:1000 in 1x Roti Block for 1h at RT.
  • Wash membrane 2x for 10min with 1x TBST.
  • Incubation with secondary
    • HRP-conjugated goat anti-rabbit IgG (H+L) (Jackson ImmunoResearch, 111-035-003) diluted 1:1000 in 1x Roti Block for 1h at RT,
    • goat anti-mouse IgG (H+L) (Jackson ImmunoResearch, 115-005-003, lot 128525) diluted 1:10000 in PBS for 45min at RT, or
    • HRP-conjugated goat anti-rat IgG (H+L) (Jackson ImmunoResearch, 112-035-003) diluted 1:10000 in PBS for 45min at RT.
  • Wash membrane 5x for 5 min with TBS-T.
  • Reveal protein bands using ECL (GE Healthcare, RPN3244V2, 9633722) on an LAS mini system (GE healthcare).
Experimental Notes

Western blot using ABIN933127 on HeLa cell extracts reveals a band of the expected molecular weight. The same band is also detected by an independent CLNS1A antibody and disappears upon siRNA knock-down.

Validation Images
Western Blotting validation image for anti-Chloride Channel, Nucleotide-Sensitive, 1A (CLNS1A) antibody (ABIN4903315) Western blot analysis of HeLa cell lysates without (HeLa) or with siRNA knock-down (H...
Immunogen A synthetic Peptide of human CLNS1A
Isotype IgG
Purification Affinity purification
Antigen
Alternative Name CLNS1A (CLNS1A Antibody Abstract)
Background This gene encodes a protein that functions in multiple regulatory pathways. The encoded protein complexes with numerous cytosolic proteins and performs diverse functions including regulation of small nuclear ribonucleoprotein biosynthesis, platelet activation and cytoskeletal organization. The protein is also found associated with the plasma membrane where it functions as a chloride current regulator. Pseudogenes of this gene are found on chromosomes 1, 4 and 6. Several transcript variants encoding different isoforms have been found for this gene.
Molecular Weight 26.215 kDa
Gene ID 1207
UniProt P54105
Pathways Ribonucleoprotein Complex Subunit Organization
Application Notes WB 1:500 - 1:1000
IHC 1:20 - 1:50
FC 1:20 - 1:50
Restrictions For Research Use only
Format Liquid
Buffer PBS with 0.02 % sodium azide, 50 % glycerol,  pH 7.3.
Preservative Sodium azide
Precaution of Use This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice Avoid repeated freeze/thaw cycles.
Storage -20 °C
Storage Comment Store at -20C. Avoid freeze / thaw cycles.
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